To develop a new Chinese medicine (CM)-based drug and to evaluate its safety and effect for suppressing acute respiratory distress syndrome (ARDS) in COVID-19 patients. A putative ARDS-suppressing drug Keguan-1 was first developed and then evaluated by a randomized, controlled two-arm trial. The two arms of the trial consist of a control therapy (alpha interferon inhalation, 50 µg twice daily; and lopinavir/ritonavir, 400 and 100 mg twice daily, respectively) and a testing therapy (control therapy plus Keguan-1 19.4 g twice daily) by random number table at 1:1 ratio with 24 cases each group. After 2-week treatment, adverse events, time to fever resolution, ARDS development, and lung injury on newly diagnosed COVID-19 patients were assessed. An analysis of the data from the first 30 participants showed that the control arm and the testing arm did not exhibit any significant differences in terms of adverse events. Based on this result, the study was expanded to include a total of 48 participants (24 cases each arm). The results show that compared with the control arm, the testing arm exhibited a significant improvement in time to fever resolution (P=0.035), and a significant reduction in the development of ARDS (P=0.048). Keguan-1-based integrative therapy was safe and superior to the standard therapy in suppressing the development of ARDS in COVID-19 patients. (Trial registration No. NCT 04251871 at www.clinicaltrials.gov)

Caveolin-1 and flotillin-1 are considered as markers of lipid rafts which can be regarded as sorting platforms for targeted transport of transmembrane proteins and are involved in fundamental cellular events such as signal transduction, cell adhesion, lipid/protein sorting, and human cancer. We addressed caveolin-1 and flotillin-1 expression in 90 human hepatocellular carcinoma (HCC) and adjacent noncancerous tissues (ANT) samples by SDS-PAGE and immunoblotting with specific antibodies. Significant caveolin-1 and flotillin-1 overexpression was found in HCC tissues compared to ANT and was confirmed by immunohistochemistry. Raft-associated Akt signaling pathway components involved in the regulation of cell survival were altered by western blotting in HCC microdomain-enriched subcellular fractions purified from paired HCC and ANT samples. Our results demonstrated that the activity of raft-associated but not total membrane Akt determines its cellular functions. Lipid rafts differ in different types of tissues, which allows for the possibility of tissue-type-specific targeting for cell survival.

There is increasing interest in the role of T follicular helper (Tfh) cells in autoimmunity from the perspective of both their role in breach of tolerance and their effects on the natural history of disease progression. Indeed, the critical role of Tfh cells in autoimmunity is further highlighted based on their location in the germinal center (GC), a pathogenic hot spot for development of autoreactivity. To address the role of Tfh cells in primary biliary cirrhosis (PBC), we comprehensively evaluated the immunobiology of CXCR5+CD4+ Tfh cells in 69 patients with PBC, including a nested subgroup of 16 autoimmune hepatitis (AIH) and 20 healthy controls (HC), followed for 1 year. We report herein several key observations. First, there was an increased frequency of circulating Tfh cells in patients with PBC compared to AIH (P < 0.05) and HC (P < 0.01). Second, the function of circulating Tfh cells from PBC patients, including interleukin (IL)‐21 production (P < 0.05), the ability to promote B‐cell maturation, and autoantibody production, were greater than HC. Third, the frequency of these cells was significantly decreased in ursodeoxycholic acid (UDCA) responders compared to UDCA‐treated nonresponders, in both cross‐sectional (P = 0.023) and longitudinal studies (P = 0.036), respectively. Indeed, similar increases of Tfh cells were noted in liver and spleen. Conclusion: These results significantly extend our understanding of lymphoid subpopulations in PBC and their relative role in disease expression. Our data also provide a novel biomarker for evaluation of the effectiveness of new therapeutic approaches. (Hepatology 2015;61:627‐638)

Objective To analyze the rate of alcohol relapse after liver transplantation (LT)for alcohol-related liver disease (ALD)and its influence on patient’s survival in a single center. Methods This retrospective study included 435 consecutive adult recipients of a primary liver graft between 2005 and 2013 in our hospital,Among 81 cases of patients with excessive alcohol consumption before transplantation,13 had ALD as primary indication,and 68 had ALD as secondary indication with other liver related diseases as their primary indications for transplantation. Medical records were reviewed, data on alcohol consumption before and after LT,and survival were collected. Results The mean follow-up time of 435 patients was 52.2 months. After transplantation,alcohol relapse rate in patients with ALD as primary indication was higher than those with ALD as secondary indication (46.15% vs 13.24 % ). Comparisons between groups were performed by Fisher’s exact test (P= 0.016). The survival rates assessed by Kaplan-Meier method were 81.4% ,100% and 85.3% at 8 years for patients with ALD as primary indication,as secondary indication and without ALD,respectively. Survival curves were compared using Log Rank statistic,and no differences were found among 3 groups(P= 0.117). Abstinence time, which was more than 6 months before LT in patients with ALD,had no influence on alcohol relapse after LT. Conclusion After transplantation,alcohol relapse rate in patients with ALD as primary indication is higher than those with ALD as secondary indication. Abstinence time,which is more than 6 months before LT in patients with ALD,has no influence on alcohol relapse after LT.

Objective To evaluate the efficacy and safety of domestic adefovir dipivoxil in the treatment of chronic hepatitis B with e antigen-positive.Methods The multicenter open clinical trial was conducted,the 1930 patients received oral adefovir dipivoxil 10 mg/d.Follow-up observations by standard operating procedures(SOP).The changes in biochemical,virological parameters and adverse reactions were observed after treatment on 12 weeks,24 weeks,36 weeks and 52 weeks.Results After 12 weeks,24 weeks,36 weeks and 52 weeks of treatment,the rates of the serum HBV DNA negative-conversion and alanine aminotransferase(ALT) renormalization were 16.42％,28.55％,37.25％,43.32％ and 29.69％,49.02％,56.42％,62.95％ respectively.After 24 weeks and 52 weeks of treatment,the rates of HBeAg negative-conversion were 12.59％,19.38％ respectively.After that,as the treatment duration prolonged,there was an increase in the serum HBV DNA negative-conversion,ALT renormalization and HBeAg negative-conversion rate resistance rate was 0.17％ during the treatment of 52 weeks period.The adverse reaction rate of 0.72％,very few cases of urine routine showed mild abnormality,no impairments of renal function or reproductive system were emerge.Conclusions Domestic adefovir dipivoxil for treating chronic hepatitis B with e antigen-positive is safe and effective.

目的 进一步评价茵兰益肝颗粒(茵陈、郁金、当归、连翘、丹参、板蓝根、拳参、淡竹叶)治疗慢性病毒性肝炎(肝胆湿热、气滞血瘀证)的疗效及安全性.方法 采用多中心、随机、双盲、茵兰益肝颗粒360例与阳性药利肝隆颗粒120例平行对照的研究方法,治疗期共12周,服药后2周、4周、8周、12周各进行1次访视,观察丙氨酸转氨酶、天冬氨酸转氨酶复常率和未反跳率,中医证候总积分变化值,以及安全性指标.结果 主要疗效指标丙氨酸转氨酶复常率试验组59.8％,对照组32.2％,两组差异有显著统计学意义(P＜0.01);次要疗效指标天冬氨酸转氨酶复常率试验组61.5％,对照组45.8％;丙氨酸转氨酶未反跳率试验组87.9％,对照组58.8％;天冬氨酸转氨酶未反跳率试验组90.5％,对照组63.2％,以上指标以及中医证候总积分变化值两组差异均有显著统计学意义(P＜0.01).安全性方面不良事件发生率试验组为0.8％,对照组为5.2％.结论 茵兰益肝颗粒治疗慢性病毒性肝炎(肝胆湿热、气滞血瘀证)具有改善肝功能,恢复丙氨酸转氨酶、天冬氨酸转氨酶,抑制丙氨酸转氨酶、天冬氨酸转氨酶反跳以及改善患者症状的作用,同利肝隆颗粒比较具有优势.

Background Acute-on-chronic hepatitis B liver failure (ACLF-HBV) is a clinically severe disease associated with major life-threatening complications including hepatic encephalopathy and hepatorenal syndrome. The aim of this study was to evaluate the short-term prognostic predictability of the model for end-stage liver disease (MELD), MELD-based indices, and their dynamic changes in patients with ACLF-HBV, and to establish a new model for predicting the prognosis of ACLF-HBV.Methods A total of 172 patients with ACLF-HBV who stayed in the hospital for more than 2 weeks were retrospectively recruited. The predictive accuracy of MELD, MELD-based indices, and their dynamic change (A) were compared using the area under the receiver operating characteristic curve method. The associations between mortality and patient characteristics were studied by univariate and multivariate analyses.Results The 3-month mortality was 43.6%. The largest concordance (c) statistic predicting 3-month mortality was the MELD score at the end of 2 weeks of admission (0.8), followed by the MELD:sodium ratio (MESO) (0.796) and integrated MELD (iMELD) (0.758) scores, AMELD (0.752), AMESO (0.729), and MELD plus sodium (MELD-Na) (0.728) scores. In multivariate Logistic regression analysis, the independent factors predicting prognosis were hepatic encephalopathy (OR=3.466), serum creatinine, international normalized ratio (INR), and total bilirubin at the end of 2 weeks of admission (OR=10.302, 6.063, 5.208, respectively), and cholinesterase on admission (OR=0.255). This regression model had a greater prognostic value (c=0.85, 95% Cl 0.791-0.909) compared to the MELD score at the end of 2 weeks of admission (Z=4.9851, P=0.0256).Conclusions MELD score at the end of 2 weeks of admission is a useful predictor for 3-month mortality in ACLF-HBV patients. Hepatic encephalopathy, serum creatinine, international normalized ratio, and total bilirubin at the end of 2 weeks of admission and cholinesterase on admission are independent predictors of 3-month mortality. Chin Med J 2012;125(13):2272-2278

Objective To study the bioavailability of the tested and reference huperzine A sustain-release tablets in healthy male volunteers.Methods According to the four-crossing design,each volunteer was orally given huperzine A sustain-release tablets or huperzine A.The plasma concentrations were determined by LC-MS/MS.Pharmacokinetic parameters were obtained using DAS 2.0 program.Results The main pharmacokinetic parameters of huperzine A tablets test and reference were as follows: Cmax were(0.75±0.25),(1.57±0.44)ng·mL-1,Tmax were(5.77±3.60),(1.27±0.98)h,AUC0–t were(14.4±5.81),(15.1±5.66)ng·h·mL-1,AUC0-∞ were(17.1±7.29),(16.7±6.77)ng·h·mL-1,in 200 μg groups respectively.The relative bioavailability was(97.2± 24.4)%.The data of the reference and tested drugs were not different as shown by F test.Conclusion The result shows that the two huperzine A tablets made in two different preparations are bioequivalent.Huperzine A sustain-release tablets clearly have the characteristic of slow releasing properties.

Objective To evaluate the efficacy and safety of Liuweiwuling tablet in the treatment of chronic hepatitis B(CHB).Methods A multicenter,randomized,double-blind,placebo-controlled trial of Liuweiwuling tablet for CHB was conducted.The trial included two treatment periods,12 weeks of treatment with regular doses and another 12 weeks of treatment with reduced doses.During the first 12 weeks,229 CHB patients were randomly divided into the test group(n=114) and the control group(n=115).The patients in the test group were administered with Liuweiwuling tablet(3 tablets per time,3 times per day) and placebo of imitating bifendate(5 pills per time,3 times per day),and the patients in the control group with placebo of imitating Liuweiwuling tablet(3 tablets per time,3 times per day) and bifendate(5 pills per time,3 times per day).In the second 12 weeks,all patients received gradually reduced doses of treatment every 4 weeks.After 24 weeks of treatment,the effective patients were given 12 weeks of follow-up.Results At week 24 of treatment,ALT normalization rate was 68.42% in the test group and 61.74% in the control group,and the difference between them was not significant.The effectiveness rate of the decrease in ALT levels was 72.81% in the test group and 66.96% in the control group,and the difference between them was not significant.It was indicated that Liuweiwuling tablet was as good as bifendate in reducing ALT levels.After 12 weeks of follow-up,ALT levels remained normal in 84.85% of the patients in the test group and in 62.30% of the patients in the control group,and the difference between them was significant.It was shown that the test group was superior to the control group in keeping ALT at a normal level.No adverse reactions were found in the test group and the control group during clinical study.Conclusions Liuweiwuling tablet is as effective as bifendate in reducing ALT levels and superior to bifendate in keeping ALT at a normal level after withdrawal from drugs.It is suggested that Liuweiwuling tablet is effective and safe in the treatment of CHB patients with elevated ALT levels,and it has persistent and stable effect on reducing ALT levels.

Objective To determine the expiry date of entecavir dispersible tablets from accelerating and long-term stability experiments.Methods According to references and under simulated packing conditions,the quality indexes of entecavir,such as shape,properties,and content determination,were periodically checked duing 18 months.Results All the detectable results conformed to regulations on quality standards.Conclusion The shelf life of dispersible tablets is defined as 18 months,as the quality is stable after they are kept for 6 months under 40℃ or under 18 months in room temperature.

Objective To establish Infectious Diseases Drug Clinical Trial Management System with self-owned intellectual property rights, so as to provide technical support for the research and development of drugs for infectious diseases. Methods According to the procedure and standards of good clinical practice (GCP), the system was constructed based on J2EE platform, Bio-Know Universal Application Platform, Struts and Hibernate technique and Oracle database. Results In the Infectious Diseases Drug Clinical Trial Management System, an effective evaluation system for infectious diseases drug clinical trials and a quality control system were established to guarantee the reliability of the management of clinical evaluation data, and an efficient personnel training system was improved to make the clinical research for infectious diseases meet the requirements of the ethics of biomedical research. The clinical trial management system could undertake the responsibility of carrying out phase Ⅰ-Ⅳ clinical trials of drugs for infec- tious diseases. Conclusions Infectious Diseases Drug Clinical Trial Management System has established successfully. Conforming to GCP standards, it provides a good platform for the evaluation of clinical trails of drugs for infectious diseases, with high practical value.

Objective To investigate the effect of ursodeoxycholic acid nanosuspension on lowering the elevated serum GGT levels in patients with HBV-related liver cirrhosis.Methods Eighty patients with HBV-related liver cirrhosis were randomly divi-ded into two groups,a treatment group and a control group.The patients in the control group received conventional liver-protecting therapy and those in the treatment group were given oral intake of ursodeoxycholic acid nanosuspension in addition to the conven-tional therapy.The serum GGT levels of the two groups were detected at week 2 and 4 of treatment.Results The differences in serum GGT levels were significant between the two groups at different time points.The serum GGT levels decreased more rapidly at week 2 and 4 of treatment in the treatment group than in the control group.Conclusion Ursodeoxycholic acid nanosuspension can lower the elevated serum GGT levels in patients with HBV-related liver cirrhosis rapidly and effectively.

Objective To establish an HPLC quantitative method for determination of entecavir in entecavir dispersible tablets.Methods HPLC was used for determining the content of entecavir with linear gradient elution.The chromatography column was C18(4.6 mm×150 mm,5 μm);The first mobile phase consisted of water-acetonitrile-trifluoroacetic acid(990∶ 10∶ 1) solution while the second mobile phase was water-acetonitrile-trifluoroacetic acid(700∶ 300∶ 1) solution.The flow rate was 1.0 ml · min-1and the detection wavelength was set at 254 nm.The column temperature was 25 ℃.Results The linear range of entecavir was 21-50 μg · ml-1(r=0.9992).The average recovery of enticavir was 99.14% and RSD was 0.15%.Conclusion This method is simple,specific and reproducible,which can be used to determine entecavir in entecavir dispersible tablets.

Objective To assess the effect of high protein and fat diet on the pharmacokinetic profiles of Huperzine A Sustained-Release Tablets by LC-MS-MS.Methods The study was conducted according to an open,randomized,2-period crossover design with a 7-day washout interval,which was administrated with a Huperzine A Sustained-Release Tablets on an empty stomach or after meal.The plasma concentrations were determined by LC-MS/MS.Pharmacokinetic parameters were obtained using DAS program.Results The major pharmacokinetic parameters of the single doses study on an empty stomach and after meal were as follows:Cmax were(0.873±0.117)ng/mL and(1.07±0.19)ng/mL,tmax were(4.6±2.3)h and(6.0±1.8)h,AUC0-t were(18.7±3.0)ng·h/mL and(19.4±3.7)ng·h/mL,AUC0-∞were(20.2±3.5)ng·h/mL and(20.6±3.9)ng·h/mL,respectively.As compared with fasting state,the relative bioavailability of fed state was determined as(97.5±10.8)%,Cmax increased and tmax delayed slightly.Conclusion This method is sensitive,convenient and proved to be suitable for clinical investigation of huperzine A pharmacokinetics.High protein and fat diet could influence Huperzine A Sustained-Release Tablets's tmax and Cmax slightly.

Objective To study the bioavailability and bioequivalence of the tested and reference Lamivudine Tablets in healthy male volun-teers.Methods According to the crossover design,each volunteer was orally given 100 mg Lamivudine Tablets.The plasma concentrations were determined by LC-MS / MS.The pharmacokinetic parameters were obtained using DAS program.Results Cmax were(1 160.954 ± 282.657) ng / mL and(1 090.206 ± 311.840) ng / mL,Tmax were(0.908 ± 0.356) h and(0.789 ± 0.240) h,t1 /2 were(3.642 ± 1.125) h and(3.360 ± 1.183) h,AUC0-t were(3 781.871 ± 684.773) ng /(mL.h) and(3 534.502 ± 798.057) ng /(mL.h),AUC0-∞ were(3 830.838 ± 692.358) ng /(mL.h) and(3 585.388 ± 790.932) ng /(mL.h),respectively.The relative bioavailability was(109.0 ± 16.2) %.The data of the reference and tested drugs were not obviously different.Conclusion The results show that the two Lamivudine Tablets made by two different corporations are bioequivalent.

Objective To investigate the stability of adefovir dipivoxil tablet and determine the expiry date.Methods Ac-cording to the Collection of Preclinical Guidelines for new Drugs,under market packing circumstances,the quality indexes of adefovir dipivoxil tablet such as shape and properties and content determination,were periodically checked in the storage time of 12 months.Results All the indexes conformed to regulations of its quality standard.Conclusion Adefovir dipivoxil tablet has high and stable quality after it is kept for 12 months in room temperature,so the shelf life of the tablet is defined as 12 months.

目的 评价头孢呋辛酯片在健康人体的相对生物利用度及生物等效性.方法 受试者随机、自身双交叉、单剂量口服受试制剂或参比制剂500 mg,采用高效液相色谱法测定血浆中药物浓度,药代动力学参数采用DAS软件处理获得.结果 两种制剂峰浓度(Cmax)分别为(5.58±1.95)μg/mL和(5.54±1.65)μg/mL,达峰时间(Tmax)分别为(2.03±0.97)h和(1.93±0.69)h.半衰期(t1/2)分别为(1.20±0.17)h和(1.19±0.13)h,0～t药时曲线下面积(AUC0-t)分别为(17.94±6.19)μg·h/mL和(17.37 ±4.73)μg·h/mL,0～∞药时曲线下面积(AUC0-∞)分别为(18.26±6.13)μg·h/mL和(17.64±4.66)μg·h/mL;相对于参比制剂,受试制剂的生物利用度为(106.54±40.59)%.受试制剂和参比制剂的Cmax,AUC0-t,AUC0-∞经对数转换后进行方差分析,两制剂间无显著性差异.结论 浙江京新药业股份有限公司研制的头孢呋辛酯片与葛兰素史克制药(苏州)有限公司生产的头孢呋辛酯片具有生物等效性.

Objective To develop an high performance liquid chromatography method for the determination of adefovir dipivoxil in adefovir dipivoxil tablets.Methods The chromatographic equipment includeds pump HP1100,inspection instrument HP1100,chemical analytical station HP,and a mobile phase composed of acetonitrile-water(40 ∶ 60) solution with 1% triethylamine and 50 mmol/L potassium dihydrogen phosphate(adjusted to pH 3.0 by phosphoric acid).The flow rate was 1.0 ml/min and monitered at 260 nm.The column temperature was room temperature.Results The calibration curves of adefovir dipivoxil showed a good linear relation over the range of 4.04-50.50 μg/ml(r=0.9999).The average recovery of adefovir dipivoxil was 99.77% and RSD was 0.24%.Conclusion This method is simple,accurate and with good reproducibility.It can be used for the determination of adefovir dipivoxil in adefovir dipivoxil tablets.

<正>多中心临床试验考虑中心效应进行疗效评价时,需参考临床界值(如等效界值或非劣效性界值)进行统计分析,但经对国内外文献检索,发现有关文献报道极少。有小部分等效性或非劣效性研究结果,基于中心效应检验无统计学差异的情况下,计量资料应用两组资料的t检验,分类资料应用两组资料的U检验,或应用均数或有效率的可信区间进行统计分析;多数文献未考虑中心效应,直接应用两组计量资料的t检验,