Study on the Effects of Guiqi Oral Liquid (归芪口服液) in Promoting Recovery of Hematopoiesis in Acute Irradiation Injured Mice

Objective: To explore the effects and possible mechanisms of Guiqi Oral Liquid (归芪口服injured mice were randomly divided into 2 groups: the treated group and the control group, and also a normal control group was set up with 6 mice in it receiving no treatment. After the mice in the former two groups were irradiated by 6.0 Gy 60Coγ-ray, every one of them was given 0.4 ml GQOL or saline in equal volume through a gastric tube twice a day for 14 days. On the 4th, 8th and 14th day after irradiation, the bone marrow mononuclear cells (BMMNC) and megakaryocytes in bone marrow tissues of the mice were counted, the proportion of hematopoietic tissues (by area) was measured, and the expression of adhesion molecules, CD44 and CD54, in bone marrow were estimated by immunochemistry. The colony forming unit of spleen (CFU-S)in the mice were counted on the 8th day after irradiation. Results: On the 4th, 8th, 14th day after irradiation,the count of BMMNC and megakaryocyte, and the proportion of hematopoietic tissues in the treated group were higher than those in the control group (P＜0.01 or P＜0.05). CD44 and CD54 expression in the treated group were higher than those in the control group on the 4th and 8th day (P＜0.01), but near normal on the 14th day ( P＜0.01). On the 8th day, CFU-S count in the treated group was higher than that in the control group (P＜0.01). Conclusion: GQOL can regulate the expression of adhesion molecules, CD44 and CD54, in the bone marrow of the acute irradiation injured mice, which may be one of the mechanisms of GQOL in accelerating the early phase hematopoiesis recovery of mice.     

Effects of Compound Zhebei Granule (复方浙贝颗粒) Combined with Doxorubicin on Expression of Specific Surface Antigens in Mice with Transplanted KG-1a Cells

Objective:To investigate the effects of Compound Zhebei Granule(复方浙贝颗粒,CZG)combined with doxorubicin hydrochloride(adriamycin,ADM)on specific surface antigens in mice with KG-1a transplanted cells.Methods:A subcutaneous tumor xenograft model was established by injection of the acute myeloid leukemia cell line KG-1a into the axillary flanks of BALB/c-nude mice.Twenty-four tumor bearing mice were divided into 4groups according to a random number table,including normal saline control group,ADM group,high-dose CZG group,and mid-dose CZG group,with 6 mice in each group.Drug administration occurred on the 14th day after cell inoculation,and normal saline control group mice were gavaged with normal saline at 0.2 m L/10 g every other day.ADM group mice were intraperitoneally injected with ADM 1 mg/kg[conversion of adults,40 mg/(m~2·d)]every other day.High-and mid-dose CZG groups mice were gavaged with CZG at the dose of 8 and 4 g/kg respectively every other day and intraperitoneally injected with ADM(1 mg/kg)every other day.The administration period lasted for10 days.The tumor xenografts were made into mononuclear cell suspensions after dissection,and the expressions of specific surface antigens,including CD34~+CD38~-,CD34~+CD38~-CD123~+,CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+,in KG-1a cell line tumor xenografts were detected by flow cytometry.Results:Compared with the control and ADM groups,expression of CD34~+CD38~-in the two CZG groups was significantly lower(P0.05).Compared with the control group,expression of CD34~+CD38~-CD123~+in the two CZG groups decreased(P0.05).The high-dose CZG group showed more obvious outcomes compared with the ADM group(P0.05).Compared with the control and ADM groups,the expression of CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+in the two CZG groups decreased(P0.05).Conclusions:CZG combined with doxorubicin could reduce the expression of CD34~+CD38~-,CD34~+CD38~-CD123~+,CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+in KG-1a cell line tumor xenografts,which shows that CZG could target leukemia stem cells and play a role in chemosensitization.     

Huangqin Decoction Attenuates DSS-Induced Mucosal Damage and Promotes Epithelial Repair via Inhibiting TNF-α-Induced NF-κB Activation

Objective:To investigate the protective effect of Chinese herbal formula Huangqin Decoction(HQD)on ulcerative colitis mouse model induced by dextran sulphate sodium(DSS)and human intestinal epithelial cell injury induced by tumour necrosis factor-α(TNF-α).Methods:In vivo,30 male C57BL/6 mice were divided into 5 groups using a random number table(n=6 per group),including control,DSS,5-aminosalicylic acid(5-ASA),HQD low-(HQD-L)and high-dose(HQD-H)groups.The colitis mouse model was established by 3%(w/v)DSS water for 5 days.Meanwhile,mice in the HQD-L,HQD-H and 5-ASA groups were administrated with 100,200 mg/kg HQD or 100 mg/kg 5-ASA,respectively,once daily by gavage.After 9 days of administration,the body weight,disease activity index(DAI)score and colon length of mice were measured,the pathological changes of colons were analyzed by hematoxylin-eosin staining(HE)staining,and the levels of serum interleukin(IL)-6,IL-1βand TNF-αwere measured by enzyme linked immunosorbent assay.In vitro,the human colon epithelial normal cells(FHC cells)were exposed to HQD(0.6 mg/mL)for 12 h and then treated with TNF-α(10 ng/mL)for 24 h.The tight junction(TJ)protein expression levels of Claudin-4 and Occludin,and the protein phosphorylation levels of p65 and inhibitor of nuclear factor kappaB(NF-κB)-α(IκBα)were measured by Western blot.Results:In vivo,compared with the DSS group,HQD-H treatment attenuated the weight loss and reduced DAI score of mice on the 8th day(P<0.05).Moreover,HQD-H treatment ameliorated the colon shortening in the DSS-induced colitis mice(P<0.05).HE staining showed HQD attenuated the pathological changes of colitis mice,and the histological scores of HQD-H and 5-ASA groups were significantly decreased compared with the DSS group(P<0.05).Meanwhile,HQD-H and 5-ASA significantly decreased the serum IL-1β,IL-6 and TNF-αlevels of mice(P<0.05).In vitro experiments showed that HQD up-regulated Occludin and Claudin-4 protein expressions and inhibited p-p65 and p-IκBαlevels in FHC cells compared with the TNF-αgroup(P<0.05).Conclusion:HQD significantly relieved the symptoms in DSS-induced colitis mice by inhibiting pro-inflammatory cytokines expression and maintained the homeostasis of TJ protein in FHC cells by suppressing TNF-α-induced NF-κB activation.     

Protective Effect of Zengye Decoction(增液汤)on Submandibular Glands in Nonobese Diabetic Mice

Objective: To investigate the protective effect of Zengye Decoction(增液汤, ZYD) on the submandibular glands(SMGs) in nonobese diabetic(NOD) mice. Methods: Twenty-seven female NOD mice were randomly equally divided into 3 groups: the model group, the hydroxychloroquine(HCQ) group, and the ZYD group. Nine C57/B6 mice served as the normal group. After 1-week acclimation, the HCQ and ZYD groups were intragastrically administered with HCQ and ZYD, respectively, and the normal and model groups were administered with normal saline. Changes in the salivary flow rate were observed. Mice from all 4 groups were sacrificed at the age of 20 weeks. The serum and SMGs were collected. Serum cytokines gamma-interferon(IFN-γ), interleukin-10(IL-10) were detected by enzyme-linked immunosorbent assay. Histological changes in the submandibular glands were examined by hematoxylin and eosin staining. The mRNA expression of IFN-γ, IL-10 and vasoactive intestinal peptide(VIP) in the submandibular glands were measured by realtime polymerase chain reaction. Results: Compared with the model group, the salivary flow of the ZYD group significantly increased(P0.05), the extent of the histological changes was ameliorated(P0.05), and the Th1/Th2 cytokine imbalance was remedied(P0.05). In the ZYD-treated mice, the VIP mRNA was up-regulated(P0.05). Conclusions: ZYD is beneficial in protecting structure and function of SMGs in NOD mice. The mechanism may be associated with the correction of the Th1/Th2 cytokine imbalance, and with the prevention of a progressive decline of the VIP level.     

Changes of Expressions of VEGF, bFGF, and Angiogenesis, and Effect of Benazepril, bFGF on Angiogenesis in Acute Myocardial Infarction Model of the Rabbits

Objective To explore the changes of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and angiogenesis, and the effects of bFGF, angiotensin converting enzyme inhibiter(ACEI) benazepril on the angiogenesis in acute myocardial infarction (AMI) model of rabbits, and to provide a probable evidence for the treatment of AMI. Methods AMI model was established by ligating anterior descending branch of coronary artery of Japan-Sino hybridization white rabbits. The postoperative rabbits were randomly divided into 6 groups and each group was treated with different drugs. Groups 1 and 2 were treated with normal saline (NS) for 28 and 14 days (d), group 3 and 4 with bFGF for 28 and 14 d, groups 5 with benazepril for 14 d, and group 6 with benazepril and bFGF for 14 d respectively. The rabbits were killed on the 14th or 28th d and their hearts were excised, sectioned and stained with HE, Masson trichrome to observe VEGF, bFGF and CD34 under a microscope, which were quantified with a computer-assisted morphometry. Results Compared with group 1, the granulation tissue of infarction zone (IZ) in group 2 freshened up, and the capillary density (CD) in IZ was increased (P=0.002). The CD in the IZ as well as VEGF and bFGF in groups 3 and 4 were increased respectively (P=0.011-0.037). In group 5 the changes of VEGF and bFGF were not found in the IZ and the border zone (BZ) while CD was significantly increased (35.4% and 25.6%, P=0.036 and 0.037). Compared with group 2, the CD in the IZ and BZ of group 6 was significantly increased (63.4% and 44.3% P=0,007 and 0.007), meanwhile VEGF and bFGF were increased. Compared with group 5, only VEGF was increased. Conclusion Intravenous bFGF may increase VEGF and bFGF significantly, thus promoting the angiogenesis in the IZ and BZ in cardiac infarction as VEGF and bFGF are the potent angiogenic growth factors. Benazepril may promote angiogenesis in the IZ and BZ in cardiac infarction, but its mechanism is irrelative to the expression of VEGF and bFGF. The combination of benazepril and bFGF may promote, to some extent, the expression of VEGF and bFGF, but their effect on angiogenesis has not been found.     

Effect of blockage of costimulatory signal on murine abortion-prone model

Background Inhibition of the key costimulatory signals results in T cell anergy, indicating the alloantigen-specific immunologic unresponsiveness. In this study, the effect of blockage of costimulatory signal CD(86) on murine abortion-prone model was studied.Methods Thirty CBA/J female mice cohabitated with DBA/2 male or BALB/c male mice were investigated. CBA/J ×DBA/2 matings were used as the abortion-prone model, and CBA/J × BALB/c matings were used as the normal pregnant model. The abortion-prone models were divided into experimental and control groups, and the normal pregnant models were set as a normal group (10 mice in each group). The mice in the experimental group were treated with anti-mouse CD(86) monoclonal antibody (mAb) (100 μg) on day 4.5 of gestation, while the controls received irrelevant-isotype matched rat IgG(2b). As for the normal group, nothing was given to the mice. The mice were killed on day 13.5 of gestation, embryo resorption rate and the expression of transforming growth factor β(1 )(TGF-β(1)), plasminogen activator inhibitor 1 (PAI-1), and matrix metalloproteinase 9 (MMP-9) were detected. Then the data were analyzed by Chi-square test and Fisher’s exact test.Results The embryo resorption rate in the experimental (8.2%) and normal groups (7.7%) was significantly lower than that of the control (23.5%, P&lt;0.05). No significant difference was detected between the experimental and normal groups (P&gt;0.05). The positive expression rates of TGF-β(1) and PAI-1 proteins in the experimental and normal groups were significantly higher than those in the control group (P&lt;0.05). The positive expression rate of MMP-9 protein in the experimental and normal groups was significantly lower than that in the control group (P&lt;0.05). No significant difference in the positive expression rates of the three proteins was detected between the experimental and normal groups (P&gt;0.05).Conclusions Blockage of costimulatory signal CD(86) at early pregnancy can treat uncertain recurrent spontaneous abortion by stimulating the expression of TGF-β(1), MMP-9 and PAI-1 and reducing the embryo resorption rate.     

CD69 expression on airway eosinophils and airway inflammation in a murine model of asthma

Background Asthma is a chronic airway disease with inflammation characterized by physiological changes （airway hyper-responsiveness, AHR） and pathological changes （inflammatory cells infiltration and mucus production）. Eosinophils play a key role in the allergic inflammation. But the causative relationship between eosinophils and airway inflammation is hard to prove. One of the reasons is lack of activation marker of murine eosinophils. We investigated the expression of CD69 on murine eosinophils in vitro, the relationship between the expression of CD69 on eosinophils from peripheral blood and bronchoalveolar lavage fluid and on airway inflammation in asthmatic mice. Methods Eosinophils from peripheral blood of IL-5 transgenic mice （NJ.1638） were purified. Mice were divided into five groups： wild type mice sensitized and challenged with saline （WS group）, wild type mice sensitized and challenged with ovalbumin （WO group）, IL-5^-/- mice sensitized and challenged with saline and transferred with purified eosinophils （ISE group）, IL-5^-/- mice sensitized and challenged with OVA and transferred with purified eosinophils （IOE group）, IL-5^-/- mice sensitized and challenged with OVA and transferred with purified eosinophils, pretreated with anti CD4 monoclonal antibody （IOE＋antiCD4mAb group）. IL-5^-/- mice were sensitized with OVA at day 0 and day 14, then challenged with OVA aerosol. On days 24, 25, 26 and 27 purified eosinophils were transferred intratracheally to IL-5^-/- mice. On day 28, blood and BALF were collected and CD69 expression on eosinophils measured by flowcytometry. Results Purified eosinophils did not express CD69. But eosinophils cultured with PMA＋MA, IFN- T, IL-5 or GM-CSF expressed CD69 strongly. Eosinophils from blood of WO, WS group did not express CD69 at all. The numbers of eosinophils in BALF of WO group, IOE group, ISE group and IOE＋antiCD4mAb group were significantly higher than in mice of WS group which did not have eosinophils at all. CD69 expression on eosinophils in BALF of IOE and WO groups was strong. Eosinophils in BALF of ISE and IOE＋antiCDmAb groups did not express CD69. The mucus production result was similar to CD69 expression. There were eosinophils infiltration in lung slides of all groups except WS group. Conclusion Activation in airway of eosinophils could directly lead to airway inflammation.     

Effect of Micardis on the Expression of Renal Medulla Aquaporin-2 in Diabetic Mice

In current study, the effect of angiotensin receptor blocker Micardis on the localization and expression of aquaporin-2 (AQP2) was investigated in the renal medullary collecting duct of mice with diabetic nephropathy (DN). Mice were divided into three groups:normal group, DN group and Micardis-treated group. Six weeks after establishment of STZ-induced DN model in mice, the expression of AQP2 in renal medulla was detected measured by semiquantitative immunofluorescence histochemistry and Western blot techniques, and the localization of AQP2 by confocal immunofluorescence laser scanning microscopy. The results showed that the urinary osmolality was decreased in DN group as compared with normal group (2.39±0.11 vs 3.16±0.16, P<0.05). Although the localization of AQP2 on the renal medulla was unchanged, the expression of AQP2 was increased significantly in DN group as compared with normal group. Micardis could partly attenuate above changes. It was concluded that treatment with Micardis could partly rectify the abnormal expression of AQP2 in renal medulla of DN mice, which suggested that rennin-angiotensin system (RAS) is implicated in the pathogenesis of DN by regulating the expression of AQP2.     

Enhancing Effects of Ligustrazine on Expression of CD31 and Hematopoietic Reconstitution in Syngenic Bone Marrow Transplantation of Mice

Summary： The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation （BMT） mice was studied. Fifty-six Balb/c mice were divided into 3 groups： normal control group. BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respecxively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th, 14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells （BMNC） was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed thai in ligustrazine treated group the peripheral blood leukocylcs, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMTcontrol group （P〈0.01 or P〈0.05）. In conclusion, ligustrazine could obviously enhance the CD31 expression on the surface of BMNC after syngcnic BMT in mice, which may be one of the mecha- nisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.     

Analysis of CD4^＋CD25^＋ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4 CD25 regulatory T cells (CD4 CD25 Treg) and Foxp3 mRNA in peripheral blood mononuclear cells (PBMCs) from patients with asthma were investigated in order to elucidate the possible roles of CD4 CD25 Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls (normal control group) and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4 CD25 Treg ratio and Foxp3 mRNA in PBMCs of exac-erbation and persistent groups were lower than that of remission and normal control groups (P<0.05). Although the CD4 CD25 Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them (P>0.05). As compared with persistent group, exacerbation group had lower CD4 CD25 Treg ratio and Foxp3 mRNA (P<0.05). It was indicated that the decrease of CD4 CD25 Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.     

Expression of CD80／CD86 and CTLA-4 mRNA in Peripheral Blood Mononuclear Cells of the Patients with Systemic Lupus Erythematosus

The role of CD80/CD86 and CTLA-4 in the pathogenesis of systemic lupus erythematosus and their clinical significance was investigated. By using RT-PCR technique, the expression of CDS0/CD86 and CTLA-4 mRNA in peripheral blood mononuclear cells (PBMC) were semiquantitatively detected in 32 patients with active SLE. The results showed that the percentage of positive CD86 expression in active SLE was increased significantly as compared with normal controls (90.63% vs 60.00 %, P＜0.01). The mean level of CD86 mRNA expression in active SLE group was markedly higher than in the normal controls (0. 6410 0. 0174 vs 0. 4510 0. 0402, P＜0. 001).Compared with normal controls, the percentage of positive CTLA-4 expression and the mean level of CTLA-4 mRNA expression in active SLE group were both increased significantly (both P＜0.01). There was no statistical differences in positive rate of CDS0 and the average level of CDS0 mRNA between the two groups (both P~0.05). It was concluded that the aberrant expression of CD86 and CTLA-4 might play an important role in the activity and development of SLE.     

The effects of ischemia-reperfusion injury and hepatic artery ischemia on CD14 expression in canine auto-transplantation livers

Objective: To study the effect of ischemia-reperfusion injury(IRI) and hepatic artery ischemia(HAI) on CD14 expression in canine auto-transplantation livers. Methods:Liver orthotopic auto-transplantation models were applied with 30 healthy male Xi’an canines which were randomly divided into a control group, simultaneous reperfusion(SR) group and HAI group. CD14 protein expression, Malonaldehyde (MDA) Contents in hepatic tissues and ALT values in serum were detected, and the pathological changes of hepatic tissues was investigated under the light microscopy. Results:The level of CD14 protein expression in SR and HAI group tended to be time-dependent and both higher than controls with statistical significance(P < 0.01); The peak values of these two groups both occurred at 4 h, but the level in HAI group (11.94± 0.43) was evidently higher than that in SR group(3.04± 0.34). MDA contents in liver tissue, ALT values in serum and pathological changes showed the same changing tendency as CD14 expression. Conclusion:① Up-regula- tion of CD14 expression may be the receptor-mechanism of Kupffer cells(KCs) activation in liver transplantation. ② HAI can up- regulate CD14 expression after portal vein reperfusion, improve the activity of KCs further more, increase OFRs production and coop- erate with portal reperfusion, and finally aggravate the grafts injury.     

Clinical Observation of Shuanghuang Shengbai Granule(双黄升白颗粒) on Prevention and Treatment of Myelosuppression Caused by Chemotherapy in Cancer Patients

Objective: To study the efficacy and safety of Shuanghuang Shengbai Granule(双黄升白颗粒, SSG), a traditional Chinese herbal medicine, on myelosuppression of cancer patients caused by chemotherapy. Methods: A total of 330 patients were randomly assigned to the treatment group(220 cases, analysed 209 cases) and the control group(110 cases, analysed 102 cases) with a 2:1 ratio by envelope method. The patients in the treatment group at the first day of chemotherapy started to take SSG for 14 days, while the patients in the control group took Leucogon Tablets. The changes of the blood routine, clinical symptoms and immune function in both groups were observed for safety and efficacy evaluation. Results: At the 7th day of chemotherapy, the white blood cells(WBCs) level in the treatment group was significantly higher than that in the control group(P0.05). After treatment, the WBCs rate in the normal range accounted for 50.2% in the treatment group, the myelosuppression of WBCs and neutrophil were mainly grade Ⅰ, while 8.1% and 5.7% of patients emerged grade Ⅲ and grade Ⅳ myelosuppression, respectively. The incidence of myelosuppression of the treatment group was significantly lower than that of the control group(P0.05). The total effective rate of Chinese medicine syndrome in the treatment group was significantly higher than that in the control group(84.2% vs. 72.5%, P0.05). The immune cell levels in both groups were maintained in the normal range. Compared with that before treatment, the levels of CD3~+ and CD4~+ cells were significantly increased in the treatment group after treatment(P0.05). The discrepancy of CD3~+ and CD4~+ cell activity before and after treatment in both groups were significantly different(P0.05). No obvious adverse event occurred in both groups. Conclusion: SSG had a protection effect on bone marrow suppression, and alleviated the clinical symptoms together with clinical safety.     

Construction and Expression of DNA Vaccine pIRES-Sj97-Sj14-Sj26 and Its Immunogenicity in Mice

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj 14-Sj26 that contains fatty binding protein （Sj 14）, GST （Sj26） and paramyocin （Sj97） that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj 14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was transfected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and plRES-Sj97-SjI4-Sj26 plasmid DNA, plRES-Sj 14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA, plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the plRES-Sj97-Sj 14-Sj26 group as compared with the plRES blank vector, normal saline and plRES-Sj26 groups （P〈 0.01） and the plRES-Sj 14-Sj26（P〈0.05）. Single splenocyte suspension was prepared to detected the level of IFN-T by ELISA and the lymphocyte stimulating index （SI） by MTT. SI was significantly higher of in the plRES-Sj97-Sj 14-Sj26 group than in other groups （P〈 0.01）, while the IFN-T level was significantly higher the plRES-Sj97-Sj 14-Sj26 group than in plRES blank vector and normal saline groups （P〈0.01）, but no significant differences were found when compared with plRES-Sj 14-Sj26 and plRES-Sj26 groups. Flow cytometery showed that the percentages of CD4＋ and CD8＋ T cells were much higher in the plRES-Sj97-Sj 14-Sj26 group （P〈 0.01, P〈0.05）. It was concluded that plRES-Sj97-Sj 14-Sj26 vaccine may induce stronger immune response in BALB/c mice.     

血必净注射液对小鼠系统性红斑狼疮的治疗作用

Objective: To observe the effects of Xuebijing Injection (血必净注射液) on dendritic cells (DCs) and T lymphocytes, and the potential mechanisms of its therapeutic effect on systemic lupus erythematosus (SLE). Methods: A widely used mouse model, SLE-prone BLLF1 mice aged 8–10 weeks, was employed. Mice were randomly divided into 4 groups: a normal group, a model group and two treatment groups treated with Xuebijing Injection with a dose of 6.4 mL/kg via intraperitoneal administration for SLE-prone BLLF1 mice aged 8 weeks (treatment A group) and 10 weeks (treatment B group). Renal tissue sections were stained with Masson''s trichrome and periodic acid-silver methenamine. Histopathological changes in the kidney were evaluated by a light microscopy. The capacity of the DCs isolated from the spleen to stimulate the T cell proliferation in response to concanavalin A (Con A) was determined. Results: Compared with the model group, levels of anti-dsDNA antibodies in the two treatment groups decreased remarkablely (P<0.01, P<0.05), and levels of serum creatinine and blood urea nitrogen increased (P<0.01, P<0.05). Pathological changes were found in the kidney in the model group. Histopathological abnormalities were alleviated in the two treatment groups. Treatment with Xuebijing injection also significantly upregulated the expression of CD80, CD86 and major histocompatibility class Ⅱ by DCs compared with the model group (P<0.05). When splenic T lymphocytes from BLLF1 mice were co-cultured with DCs at ratios of 1:100, 1:150 and 1:200 for 3 and 5 days, the proliferation of T lymphocytes was suppressed compared with the normal group (P<0.05), but this was restored by Xuebijing Injection under the same conditions. In the model group, levels of tumor necrosis factor (TNF)-α in supernatants were significantly elevated compared with the normal group (P<0.01), interleukin-2 levels decreased (P<0.05), while these changes were significantly alleviated in the Xuebijing treatment groups. Conclusions: Xuebijing Injection alleviated renal injury in SLE-prone BLLF-1 mice. The mechanism might be through influencing T cell polarization mediated by DCs, and Xuebijing Injection might be a potential drug that suppresses immune dysfunction in patients with SLE.     

Study on the correlation of tongue manifestation with fibrinogen and neutrophil in acute cerebral infarction patients

<正>Objective:To investigate the correlation of tongue manifestation with the fibrinogen level and the neutrophil count in blood of acute cerebral infarction patients.Methods:A total of 200 patients with first unilateral cerebral infarction in Neurology Department of Xuanwu Hospital from March,2008 to February,2009 were recruited in this study.The correlation of the tongue fur color and texture with the blood fibrinogen level and the neutrophil count was analyzed in these patients.Results:The level of fibrinogen and neutrophil count in thick fur group were significantly higher than that in thin fur group(P0.05).There was no statistical difference in the level of fibrinogen and neutrophil count found between moist fur and dry fur.Statistical significance existed in the level of fibrinogen between the greasy tongue fur group and non-greasy tongue fur group(P0.05).The level of fibrinogen and the neutrophil count were compared among different fur color groups,revealing that the level of fibrinogen in yellowish fur group was higher than that of white fur group and normal value with statistical significance(P0.05) with neutrophil count in yellowish fur group being significantly higher than that in white fur group.Conclusion:Our results suggested that the change of tongue manifestation was associated with the level of fibrinogen and the neutrophil count in the blood of cerebral infarction patients.     

Xuebijing Injection (血必净注射液) and Resolvin D1 Synergize Regulate Leukocyte Adhesion and Improve Survival Rate in Mice with Sepsis-Induced Lung Injury

Objective: To investigate the effect of combined application of Xuebijing Injection(血必净注射液, XBJ) and resolvin D1(RvD1) on survival rate and the underlying mechanisms in mice with sepsisinduced lung injury. Methods: The cecal ligation and puncture(CLP) method was used to develop a mouse sepsis model. Specific pathogen free male C57 BL/6 mice were randomly divided into 5 groups(n=20 each): sham, CLP, CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1. After surgery, mice in the CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1 groups were given XBJ(25 μL/g body weight), RvD1(10 ng/g body weight), and their combination(the same dose of XBJ and RvD1), respectively. In each group, 12 mice were used to observe 1-week survival rate, while the rest were executed at 12 h. Whole blood was collected for flow cytometric analysis of leukocyte adhesion molecules CD18, lung tissues were harvested for observing pathological changes, and testing the activity of myeloperoxidase(MPO) and the expression of intercellular cell adhesion molecule 1(ICAM-1). Results: Compared with the CLP group, the histopathological damage of the lung tissues was mitigated, MPO activity was decreased in the CLP+XBJ and CLP+RvD1 groups(P0.05). In addition, the 1-week survival rate was improved, proportion of CD18-expressing cells in whole blood and ICAM-1 protein expression in lung tissue were decreased in the CLP+XBJ+RvD1 group(P0.05 or P0.01). Conclusion: XBJ together with RvD1 could effectively inhibit leukocyte adhesion, reduce lung injury, and improve the survival rate of mice with sepsis.     

Expression and Significance of Toll-like Receptor 2, 4 of Peripheral Blood Mononuclear Cells in Acute Abdomen Patients Associated with Systemic Inflammatory Response Syndrome

The changes of Toll-like receptor (TLR) 2, 4 of peripheral blood mononuclear cells (PBMCs) in the acute abdomen patients associated with systemic inflammatory response syndrome (SIRS) and their potential significance were explored. A clinical study was performed on 103 acute abdomen patients in whom 65 were associated with SIRS. Forty healthy individuals served as normal controls. The mRNA expression of TLR2, 4 was detected by RT-PCR, and the expression of TNF-αand EL-6 by ELISA. The level of plasma endotoxin, hospital stay and mortality were measured. It was found that the endotoxin level was increased to varying degrees in all the acute abdomen patients, and the endotoxin level was and hospital stay longer in SIRS group than in non-SIRS group (P<0.01). TLR2 mRNA, TLR4 mRNA, IL-6 and TNF-αcould be detected with low value in normal controls, but they were up-regulated markedly on the 1st day after admission. Then TLR4 mRNA, IL-6 and TNF-αwere decreased gradually, but TLR2 mRNA maintained at a high level till the 5th day. These indexes above in SIRS group were higher than those in non-SIRS group (P<0.01). The results of correlation analysis revealed the expression of TLR2, 4 mRNA was positively correlated with the levels of TNF-αand IL-6, and the hospital stay. The results of Logistic regression demonstrated that over-expression of TLR2, 4 mRNA might result in higher risk of multiple organ dysfunction syndrome (MODS). It was concluded that in the acute abdomen patients associated with SIRS, the expression of TLR2, 4 in PBMCs was increased markedly, suggesting that TLR might play an important role in the pathogenesis of acute abdomen associated with SIRS.     

Effect of Kangquan Recipe (康泉方) on BAMBI Expression in Hypothalamic-Pituitary-Prostate in Rats with Benign Prostatic Hyperplasia

Objective: To investigate the effect of Kangquan Recipe (康泉方, KQR) on bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) expression and its mechanism in rats with benign prostatic hyperplasia (BPH). Methods: Forty-eight male Sprague-Dawley rats were divided into 6 groups using a random number table, with 8 in each group: the normal group (normal saline 10 mL/kg), the model group (normal saline 10 mL/kg), the finasteride group (0.5 mg/kg), the low-dose KQR group (3.5 g/kg), the middle-dose KQR group (7 g/kg), and the high-dose KQR group (14 g/kg). The 40 rats were subcutaneously injected with testosterone propionate after castration for 30 days to establish the BPH rat model except for those in the normal group. At the same time, the corresponding drugs were administered by gavage for 30 consecutive days. The effects of different doses of KQR on the protate wet weight, prostate volume and prostate index (PI) were observed. The changes in histopathology were monitored with hematoxylin-eosin staining. BAMBI protein and mRNA expression contents were determined by Western blot and quantitative real-time polymerase chain reaction, respectively. Results: All doses of KQR could decrease prostatic epithelial tissue proliferation. Compared to the model group, the high and middle-dose KQR significantly reduced prostate wet weight, prostate volume and PI; increased BAMBI protein expression in the hypothalamus, pituitary and prostate tissue; all doses of KQR up-regulated BAMBI mRNA expression in serum, prostatic fluid and prostate tissue (P<0.05 or P<0.01). Conclusions: KQR could inhibit the proliferation of rat prostatic tissue, promote BAMBI protein expression in the hypothalamic-pituitary-prostate of rats with BPH; and increase BAMBI mRNA expression in the blood, prostatic fluid and prostate tissue of rats with BPH, showing a dose-effect relationship. KQR can be used as a potential drug for the treatment of BPH.