Simultaneous Determination of Six Compounds in Rat Plasma by Ultra‑Performance Liquid Chromatography with Tandem Mass Spectrometry: Application in the Pharmacokinetic Study of Qing Gan‑Shu Yu‑Fang

A rapid and high selective ultra?performance liquid chromatography (UPLC) with tandem mass spectrometry method for simultaneous determination of six compounds including albiflorin, paeoniflorin, picroside I, picroside II, saikosaponin A, and saikosaponin D in rat plasma was developed and validated using butyl p-hydroxybenzoate as an internal standard. One-step direct protein precipitation with acetonitrile was used to extract the compounds from the rat plasma samples. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 µm) at a flow rate of 0.4 mL/min, using gradient mode containing 0.1% formic acid in water and acetonitrile were used as the Mobile phase A and B. Electrospray ionization in negative ion mode and multiple reaction monitoring were used to identify and quantify active components. Calibration curves showed good linearity (R2 > 0.9908) over a wide concentration range for all compounds. The intra- and interday precision (relative standard deviation) ranged 2.4%–7.0% and 2.6%–8.0%, respectively. The accuracy (relative error) was from ?13.0% to 13.2% at all quality control levels. The recovery ranged from 81.1% to 92.5%. The validated method was successfully applied to pharmacokinetic study in rats after oral administration of Qing Gan?Shu Yu?Fang. The results show that one can draw a conclusion that these six active ingredients can be quickly absorbed and play a pharmacodynamic role rapidly in vivo.     

Sequential Therapy Based on Evolvement of Patterns: A New Model for Treatment of Alzheimer’s Disease

In order to solve the problem of long-term (>9 months) efficacy in the treatment of Alzheimer''s disease (AD) by conventional therapy (CT), a staged and multiply-targeted sequential therapy based on the evolvement of patterns (STEP) was developed. Its main innovations include: (1) the time order of evolution of patterns defined by Chinese medicine (CM) in AD was found, that is, "the orderly pattern evolution starting from Shen (Kidney) deficiency, progressing to phlegm, stasis and fire, and worsening to severe toxin as well as functional collapse"; (2) the cascade hypothesis of Shen deficiency in AD and its sequential therapy based on Shen-reinforcing was proposed, that is, "reinforcing Shen in the early stage and throughout the whole process, resolving phlegm, activating blood and purging fire in the middle stage, detoxifying and replenishing vitality to stop the collapse in the advanced stage", and through meta-analysis, clinical drug use was optimized, thus the leap from "inferential selection" to "evidence-based selection" was realized; (3) the STEP regimen combined with CT maintained cognitive and behavioral stability in AD patients for at least 12 months, with cognitive enhancement and behavioral synergy after 9 months, and cognitive benefit was superior to CT at 9, 12, 15, 18, 21, and 24 months, respectively. The 2-year cognitive improvement rate was increased by 25.64% (P=0.020) and the cognitive deterioration rate was decreased by 48.71% (P=0.000). Among them, the cognitive and functional benefits of Shen-reinforcing therapy for very early AD (350 cases) for 1 year were better than the placebo (P<0.001), and the dementia conversion rate was reduced by 8.85% (P=0.002). The behavioral symptomatic relief of patients with vascular dementia received fire-purging therapy (540 cases) was superior to those received CT (P=0.016). These data suggested that the STEP regimen has synergistic effects on CTs at least in terms of cognitive benefit, and the earlier the use, the greater the benefit will have. Therefore, the STEP regimen should be considered as one of the clinical options, particularly for the dearth of effective pharmaceutical or immunological interventions that are currently available for AD.     

表雄酮生产中高熔点副产物的研究

用惕告吉宁生产表雄酮(1)时,曾得到一种熔点较高、薄层色谱中极性较1大的副产物.红外、紫外光谱、色谱和制备衍生物等方法确定该化合物是3β-羟基-16-孕甾烯-20-酮肟。试验证明它系重排反应中水份偏高而生盛,该副产物可通过酸性水解、乙酰化、肟化而回收2。     

Feasibility of establishing cerebral ischemia models by using aerocyst-blocking bilateral ascending pharyngeal artery of piglets Imaging assessment

BACKGROUND: The cerebral ischemia and ischemia/reperfusion animal models are used to simulate the human cerebrovascular diseases is one of the popular topics of neurological science recently. To study the pathophysiology, pathogenesis, prophylaxis and treatment of ischemic cerebrovascular diseases and to establish the ideal animal model that is the most similar to the human cerebral ischemia, are the topics that the people generally cared about. OBJECTIVE: To evaluate the effects of aerocyst-blocking bilateral ascending pharyngeal artery on the establishment of cerebral ischemia models by using digital subtraction angiography (DSA), magnetic resonance diffusion-weighted imaging (DWI) and magnetic resonance perfusion-weighted imaging (PWI). DESIGN: Repetitive measure animal experiment. SETTING: Zhongshan Hospital Affiliated to Dalian University. MATERIALS: The experiment was carried out in the Animal Laboratory (Provincial Laboratory), Zhongshan Hospital of Dalian Univeristy from January to May 2006. A total of 14 domestic piglets, of 6 months old, weighing 12–15 kg, of either gender, were selected from Animal Experimental Center, Dalian University. Multistar T.O.P digital subtraction angiography machine was provided by Siemens Company, German. METHODS: Aerocyst-blocking bilateral ascending pharyngeal artery was used to establish cerebral ischemia models. And then, Multistar T.O.P. DSA was used for imaging of cerebral vessels before blocking, during blocking and at 0.5 and 2 hours after ischemia perfusion. GE Signa 1.5 T supraconduction magnetic resonance imaging was used for DWI examination; in addition, PWI was used based on focal sites and areas. Otherwise, magnetic resonance imaging (MRI) was used to detect signal changes of T1WI and T2WI in ischemic areas. MAIN OUTCOME MEASURES: Analytic results of DSA, DWI, PWI and MRI. RESULTS: All 14 experimental piglets were involved in the final analysis. ① DSA: The blood flow of bilateral ascending pharyngeal arteries and its branch were blocked at blocking phase, which restored 0.5 and 2 hours after reperfusion. ② DWI and PWI: There were no observable abnormalities in PWI and DWI at pre-blocking. Abnormal increased signals were found on both DWI and PWI at during and post-blocking. There were reduction in ADC and rCBF and delay in rTTP at all time points except pre-blocking. ③ MRI: There were no abnormal signals observable at any time of pre- and post-blocking in T1WI and T2WI. CONCLUSION: It is feasible to establish this kind of animal experimental models, and it can simulate the ischemic state; meanwhile, the existence and extent can be showed directly by DSA, DWI, and PWI.     

原发性下肢静脉曲张342例手术并发症分析

原发性下肢静脉曲张是外科常见病,通常均以手术治疗,行大隐静脉高位结扎,分段切除或剥脱术。手术比较小、基层已普及,但合并症较多。本文总结1980年—1990年10年间342例手术病案,分析如下: 临床资料一般资料:本组342例中,男239例、女163例,男女之比为2.4:1,年龄最小21岁,最大69岁,     

Clinical observation of Qi deficiency syndrome in 72 patients with chronic obstructive pulmonary disease treated with Yiqi Mianyi Granule

Seventy-two patients of chronic obstructive pulmonary disease (COPD) of Qi deficiency syndrome with abnormal immune indices were treated with Yiqi Mianyi Granule (YQMYG) and the efficacy was compared with that of 30 cases treated with Zhenqi fuzheng Granule (ZQFZG) for control. Results showed that the markedly effective rate of symptomatic improvement of Qi deficiency in YQMYG group was 65.3%, the total effective rate 93.1%. 88.6% of the immune indices lower than normal were corrected and 43. 7% of them were normalized, while for indices that were higher than normal the rate were 78.2% and 52.9% respectively. These results suggested that YQMYG could improve symptom of Qi Deficiency markedly, strengthen cellular immunity and regulate immune dysfunction. Its therapeutic efficacy was obviously superior to ZQFZG (P<0.Q5).     

PRENATAL DIAGNOSIS OF CONJOINED TWINSWI''TH REAL-TIME ULTRASOUNDA CASE REPORT

Conjoined twins case is a rare abnormality.The most common type of union is thoracopaguswhich is found in 75To of the cases. Conjoinedtwins have a sex predilection with 75Vo beingFcmale.i A case of conjoined twins was diagnoscdwith real-time ultrasonic scanner (Linear arraymodel, transducer 3.5 mHz) at the 21st week ofpcstation. Final.ly. it was shown late.r by po.slmor-tem examination after delivery that concurrentmalformation in head, hearts and aorta were pre-sent as previously seen on the ultrasonic imagcs.     

Neuroprotective effect of estrogen after chronic spinal cord injury in ovariectomized rats

BACKGROUND: At present, there is still lack of effective drugs for chronic spinal cord injury, whereas it is found recently that estrogen has a neuroprotective effect on brain and spinal cord injuries. OBJECTIVE: To observe the effect of estrogen on the apoptosis of nerve cells after gradual chronic spinal cord injury in ovariectomized rats. DESIGN: A randomized controlled animal trial. SETTING: Institute of Orthopaedics, the Second Hospital of Lanzhou University. MATERIALS: Sixty-five female Wistar rats of common degree, weighing 220–250 g, were provided by the experimental animal center of Lanzhou University. The rats were randomly divided into sham-operated group (n =5), estrogen-treated group (n =30) and saline control group (n =30), and the latter two groups were observed at 1, 3, 7, 14, 28 and 60 days respectively, and 5 rats for each time point. METHODS: All the rats were treated with bilateral oophorectomy 2 weeks before the experiment. T10 vertebral lamina was revolved into using plastic screw. The spinal canal impingement was not induced initially. After that, the original incision was opened to expose the screw every 7–10 days. MAIN OUTCOME MEASURES: The apoptosis and Caspase-3 positive cells in the damaged spinal cord were detected using terminal deoxynucleotidal transferase-mediated dUTP-biotin nick end labeling (TUNEL) method and Caspase-3 immunohistochemical staining at 1, 3, 7, 14, 28 and 60 days after chronic spinal cord injury respectively. RESULTS: Totally 65 rats were used, and the deleted ones during the experiment were supplemented by others. Changes of Caspase-3 expression after spinal cord injury: In the sham-operated group, only a small amount of Caspase-3 proteins were observed in the rat spinal cord, mainly located in motor neurons of spinal cord anterior horn. In the estrogen-treated group and saline control group, positive cells expressed occasionally at 1 day postoperatively, began to increase obviously at 7 days after injury, strongly expressed at 14 and 28 days, but decreased at 60 days, mainly located in the neurons of spinal cord gray matter anterior horn, and they expressed fewer in the motor neurons and white matter of ventral horn, and there were obvious differences between the estrogen-treated group and saline control group at 7, 14, 28 and 60 days (P < 0.05). CONCLUSION: Estrogen can reduce the apoptosis of nerve cells and promote the recovery of neurological function following gradual chronic spinal cord injury.