The potential mechanism of Bupleurum against anxiety was predicted by network pharmacology study and molecular docking

Metabolic Brain Disease - Bupleurum chinense DC. (Chaihu) is a traditional Chinese medicine (TCM) used in the treatment of anxiety. But the anxiolytic mechanisms of bupleurum are still unclear....     

大鼠自体异体表皮细胞悬液混合移植的实验研究

目的　探讨自、异体表皮细胞悬液混合移植技术在创面修复中的应用。　方法　 30只大鼠随机配成 15对后 ,分成细胞悬液移植组 (A组 ,10对 )和细胞膜片移植组 (B组 ,5对 )。取每只大鼠全厚皮 ,分离表皮细胞 ,并根据配对情况按 1∶1的细胞比例混合 ,体外常规培养。 4d后收获A组混合细胞悬液 ,14d后收获B组混合细胞膜片。将此细胞悬液和膜片分别转移至A、B组相应供体大鼠的去全厚皮创面。随后A组每对大鼠的创面交叉覆盖配对方的异体全厚皮 ;B组创面覆盖胶原膜及“优妥”敷料。比较移植后 2～ 3周两组的创面修复情况。　结果　术后 2～ 3周 ,A组创面大多愈合 ,表面光滑 ,与皮下连接紧密。术后第 5天 ,B组创面部分细胞膜片脱落 ,部分成活 ,膜片成活的创面后期再次出现小创面 ,经久不愈。　结论　自、异体表皮细胞悬液混合移植是一种可行的、体内构建皮肤、修复创面的方法。     

人三叶因子2治疗大鼠胃溃疡的效果评价

目的探讨人三叶因子2（hTFF2）治疗大鼠胃溃疡的疗效。方法将48只雄性Wistar大鼠随机分为Ⅰ、Ⅱ、Ⅲ组，冰乙酸法制作慢性胃溃疡模型。造模时Ⅰ、Ⅱ、Ⅲ组于胃黏膜下分别注射pcDNA3．1-hTFF2（人三叶因子2插入pcDNA3．1载体；pcDNA3．1为真核表达载体，有进入胃黏膜下细胞的特性）西米替丁及pcDNA3．1。造模后7、14d各组分别处死8只大鼠，测定溃疡面积、胃液总酸度及黏液糖蛋白水平。结果Ⅰ、Ⅱ组较Ⅲ组溃疡面积明显缩小，Ⅱ组较Ⅰ、Ⅲ组胃总酸度明显降低，Ⅰ组较Ⅱ、Ⅲ组黏液糖蛋白量明显增加，P均〈0．001。结论pcDNA3．1-hTFF2单次局部注射可通过增加黏液糖蛋白的分泌促进大鼠胃溃疡愈合。     

大鼠实验性胃溃疡自愈期间肠嗜铬样细胞的变化

目的:探讨肠嗜铬样细胞与大鼠实验性胃溃疡自愈的关系. 方法:通过制备大鼠乙酸性胃体溃疡模型,采用免疫组化、逆转录聚合酶链反应(RT-PCR)和放射免疫测定的方法,观察溃疡自愈过程中大鼠胃黏膜内ECL细胞形态、组氨酸脱羧酶(HDC)mRNA和组胺含量的变化. 结果:胃溃疡大鼠胃黏膜内HDC阳性细胞率和胞质平均灰度值在制模术后第1日即开始降低,术后第6日达到最低,随后开始回升,术后第12日基本正常. 胃黏膜内HDC mRNA表达在制模术后第1日开始降低,第3日最明显,第6日开始恢复,第9日后接近正常. 制模术后胃黏膜内组胺含量也出现下调,以术后第6日为最低. 结论:ECL细胞可通过减弱其合成组胺的功能,参与胃溃疡自愈的调节过程.     

aVR导联ST段抬高在预测非ST段抬高型急性心肌梗死患者预后中的价值

目的探讨aVR导联ST段抬高在预测首次非ST段抬高型急性心肌梗死患者短期预后中的价值。方法分析426例非ST段抬高型急性心肌梗死患者入院心电图。结果aVR导联无ST段抬高(n=281)、抬高0.05~0.1mV(n=68)和抬高≥0.1mV(n=77)患者的住院死亡率分别是1.8%、7.4%和15.6%。调整基线预测因子和入院时ST段压低的影响,aVR导联ST段抬高0.05~0.1mV和抬高≥0.1mV患者死亡的优势比分别是4.2(95%可信区间为1.4~13.5;P<0.001)和6.1(95%可信区间为2.4~17.3;P<0.001)。住院期间复发心肌缺血事件和心力衰竭发生率随aVR导联ST段抬高程度增加而增加,而不同程度aVR导联ST段抬高患者血清肌酸激酶和肌酸激酶同工酶相似。aVR导联无ST段抬高、抬高0.05~0.1mV和抬高≥0.1mV患者左主干或3支血管病变发生率分别为16.9%、37.1%和56.2%(P<0.001)。结论首次非ST段抬高型急性心肌梗死伴aVR导联ST段抬高患者预后较差,而这种差的预后与严重的冠状动脉病变有关,对这些患者进行早期介入治疗也许有重要的益处。     

Gastrointestinal hormone abnormalities and G and D cells in functional dyspepsia patients with gastric dysmotility

AIM: To investigate the relationship between gastric dysmotility, gastrointestinal hormone abnormalities, and neuroendocrine cells in gastrointestinal mucosa in patients with functional dyspepsia (FD). METHODS: Gastric emptying was assessed with solid radiopaque markers in 54 FD patients, and the patients were divided into two groups according to the results, one with delayed gastric emptying and the other with normal gastric emptying. Seventeen healthy volunteers acted as normal controls. Fasting and postprandial plasma levels and gastroduodenal mucosal levels of gastrointestinal hormones gastrin, somatostatin (SS) and neurotensin (NT) were measured by radioimmunoassay in all the subjects. G cells (gastrin-producing cells) and D cells (SS-producing cells) in gastric antral mucosa were immunostained with rabbit anti-gastrin polyclonal antibody and rabbit anti-SS polyclonal antibody, respectively, and analyzed quantitatively by computerized image analysis. RESULTS: The postprandial plasma gastrin levels, the fasting and postprandial plasma levels and the gastric and duodenal mucosal levels of NT were significantly higher in the FD patients with delayed gastric emptying than in those with normal gastric emptying and normal controls. The number and gray value of G and D cells and the G cell/D cell number ratio did not differ significantly between normal controls and the FD patients with or without delayed gastric emptying. CONCLUSION: Our findings suggest that the abnormalities of gastrin and NT may play a role in the pathophysiology of gastric dysmotility in FD patients, and the abnormality of postprandial plasma gastrin levels in FD patients with delayed gastric emptying is not related to the changes both in the number and gray value of G cells and in the G cell/D cell number ratio in gastric antral mucosa.     

Establishment of mice model with human viral hepatitis B

AIM:To establish a mice model of hepatitis B by using HBV-transgenic mice, and to transfer HBV-specific cytotoxic T lymphoo/tes (CTL) induced from syngeneic BALB/c mice immunized by a eukaryotic expression vector containing HBV complete genome DNA.METHODS: HBV DNA was obtained from digested pBR3222HBV and ligated with the vector pcDNA3. Recombinant pcDNA3-HBV was identified by restriction endonuclease assay and transfected into human hepatoma cell line HepG2 with lipofectin. ELISA was used to detect the expression of HBsAg in culture supernatant, and RT-PCR to determine the existence of HBV PreS1 mRNA.BALB/c mice were immunized with pcDNA3-HBV or pcDNA3 by intramuscular injection.ELISA was used to detect the expression of HBsAb in serum. MTT assay was used to measure non-specific or specific proliferation ability and specific killing activity of spleen lymphocytes. Lymphocytes from immunized mice were transferred into HBV-transgenic mice (2.5&#215;10^7 per mouse).Forty-eight hours later,the level of serum protein and transaminase was detected with biochemical method,liver and kidney were sectioned and stained by HE to observe the pathological changes.RESULTS: By enzyme digestion with Eco RI, Xho I and Hind Ⅲ,the recombinant pcDNA3-HBV was verified to contain a single copy of HBV genome,which was inserted in the positive direction.HepG2 cells transfected with the recombinant could stably express PreS1 mRNA and HBsAg.After immunized by pcDNA3-HBV for 4 weeks,HBsAb was detected in the serum of BALB/c mice. The potential of spleen lymphoo/tes for both non-specific and specific proliferation and the specific killing activity against target cells were enhanced. The transgenic mice in model group had no significant changes in the level of serum protein but had an obvious increase of ALT and AST. The liver had obvious pathological changes, while the kidney had no evident damage.CONCLUSION:A eukaryotic expression vector pcDNA3-HBV containing HBV complete genome is constructed successfully. HepG2 cells transfected with the recombinant can express PreS1 mRNA and HBsAg stably.Specific cellular immune response can be induced in mice immunized by pcDNA3-HBV.A mice model of acute hepatitis with HBV has been established.     

RNAi screening with shRNAs against histone methylation-related genes reveals determinants of sorafenib sensitivity in hepatocellular carcinoma cells

Sorafenib is the first drug currently approved to treat advanced hepatocellular carcinoma (HCC). However, very low response rate and acquired drug resistance makes rare patients benefit from sorafenib therapy, therefore it is urgent to find biomarkers for sorafenib sensitivity. Histone modifications, including histone methylation, have been demonstrated to influence the initiation and progression of HCC. It is of great interest to elicit the possibility whether histone methylation plays a role in regulation of sorafenib sensitivity. In present work, a high throughput RNAi screening with 176 shRNA pools against 88 histone methyltransferases (HMTs) and histone demethyltransferases genes was applied to HepG2 cells. Silencing of 3 genes (ASH1L, C17ORF49 and SETD4) was validated to specifically promote HepG2 cells sensitivity to sorafenib. Western blotting results showed that those 3 HMT genes knockdown alone or sorafenib treatments alone both induce AKT/ERK activation. However, combination treatment with sorafenib and silencing of C17ORF49 or SETD4 downregulated AKT phosphorylation and hence induced HCC cells death. Our work may provide potential biomarkers for sorafenib sensitivity and therapeutic combination for sorafenib treatment in HCC patients.     

The nonpeptide AVE0991 attenuates myocardial hypertrophy as induced by angiotensin II through downregulation of transforming growth factor-β1/Smad2 expression

The nonpeptide AVE0991 is expected to be a putative new drug for cardiovascular diseases. However, the mechanisms for the cardioprotective actions of AVE0991 are still not fully understood. We planned to determine whether AVE0991 attenuates the angiotensin II (AngII)-induced myocardial hypertrophy and whether these AVE0991 effects involved transforming growth factor β1 (TGF-β1) and Smad2. A rat model of neonatal myocardial hypertrophy was induced by AngII. The AngII group significantly increased in protein content, surface area, and [³H]leucine incorporation efficiency by cardiomyocytes, compared to those of the control group (P < 0.01). The AngII group also had elevated TGF-β1 and Smad2 expression (P < 0.01). These AngII-induced changes were significantly attenuated by AVE0991 in a dose-dependent manner. In our study, these actions of AngII (10⁻⁶ mol/l) were significantly inhibited by both concentrations of AVE0991 (10⁻⁵ mol/l and 10⁻⁷ mol/l). Moreover, the high AVE0991 group had significantly better inhibition of myocardial hypertrophy than the low AVE0991 group. Meanwhile, the beneficial effects of AVE0991 were completely abolished when the cardiomyocytes were pretreated with Ang-(1–7) receptor antagonist A-779 (10⁻⁶ mol/l). These results suggested that AVE0991 prevented AngII-inducing myocardial hypertrophy in a dose-dependent fashion, a process that may be associated with the inhibition of TGF-β1/Smad2 signaling.     

Zinc,Cadmium and Lead Accumulation and Characteristics of Rhizosphere Microbial Population Associated with Hyperaccumulator <Emphasis Type="Italic">Sedum Alfredii</Emphasis> Hance Under Natural Conditions

A field survey was conducted to study the characteristics of zinc, cadmium, and lead accumulation and rhizosphere microbial population associated with hyperaccumulator Sedum alfredii Hance growing natively on an old lead/zinc mining site. We found significant hyperaccumulation of zinc and cadmium in field samples of S. alfredii, with maximal shoot concentrations of 9.10–19.61 g kg⁻¹ zinc and 0.12–1.23 g kg⁻¹ cadmium, shoot/root ratios ranging from 1.75 to 3.19 (average 2.54) for zinc, 3.36 to 4.43 (average 3.85) for cadmium, shoot bioaccumulation factors of zinc and cadmium being 1.46–4.84 and 7.35–17.41, respectively. While most of lead was retained in roots, thus indicating exclusion as a tolerance strategy for lead. Compared to the non-rhizosphere soil, organic matter and total nitrogen and phosphorus content, CEC and water extractable zinc, cadmium, and lead concentration were significantly higher, but pH was smaller in rhizosphere soil. The rhizosphere soil of S. alfredii harbored a wide variety of microorganism. In general, significantly higher numbers of culturable bacteria, actinomycetes, and fungi were found in the rhizosphere compared to bulk soil, confirming the stimulatory effect of the S. alfredii rhizosphere on microbial growth and proliferation. Analyses of BIOLOG data also showed that the growth of S. alfredii resulted in observable changes in BIOLOG metabolic profiles, utilization ability of different carbon substrates of microbial communities in the rhizosphere soil were also higher than the non-rhizosphere, confirming a functional effect of the rhizosphere of S. alfredii on bacterial population.