Objective:To investigate the expressive characters and their significance by detecting the expressions of HBsAg,Fas and proliferating cell nuclear antigen(PCNA) in both hepatocellular carcinoma(HCC) and their surrounding non-cancerous tissues.The correlation between HBV infection and HCC,and the possible mechanism of hepatocellular carcinogenesis were discussed.Methods:HBsAg,Fas and PCNA were detected by immunohistochemistry from 52 cases of hepatocellular carcinoma as well as 42 corresponding non-cancerous tissues.Clinical data suchas serum AFP levels was also analysed.Results:Both HBsAg and Fas were strongly expressed in non-cancerous tissues,with positive rate of each achieving 78.6%.However their expression rates in cancer tissues were 21.2% and 30.8% respectively,significantly lower and less intensive than that of their corresponding non-cancerous ones(P

AIM　Diclofenac potassium sustained-release tablets were developed as a gel matrix for using twice daily and evaluated in vitro release characteristics. METHODS　The formulations were screened according to the designed in vitro release rate by the use of HPMC as the gel matrix and the hydrophobic retarding agent to modify the release. RESULTS　The release was properly characterized by the diffusion mechanism, influenced by pH of the media, slightly affected by the basket rotating speed, without the influences from the pressure exerted during the tabletting procedure. CONCLUSION　The selected formulation of diclofenac potassium sustained-release tablets could ensure the desired in vitro release rate. In addition, it proved a good manufacturing reproducibility.

Objective To study paraquat adsorbability of different field soils in Guangxi province of China. Methods HPLC method was adopted to measure the peak area of paraquat in three different media of four types of soils. Chromatographic column was Kromasil C18 column (4. 6 mm×200 mm, 5 μm); mobile phase was acetonitrile-water (including 0. 03 mol·L-1 sodium heptanesulfonate and 0. 24 mol·L-1 phosphoric acid) at a ratio of 397 (pH adjusted to 2. 0 by triethylamine). Detection wave length was 258 nm; column temperature was 25 ℃; the injection volume was 20 μL; flow rate was 0. 8 mL·min-1 . The peak areas of paraquat before and after being adsorbed were compared to calculate the adsorption rate of paraquat in different soils. Results All tested soil samples possessed the adsorption rate of paraquat over 99. 0%. Conclusion Four common field soils in Guangxi province can be used as temporary effective absorbents for the first-aid of paraquat poisoning.

Objective:To investigate the roles of donor alveolar maerophages and the recipient circulating neutrophils in early-stage reperfusion injury of lung allograft,and to study the interaction between the 2 kinds of cells.Methods:Twenty pairs of size-and weight-matched adult mongrel dogs were randomly assigned to 4 groups:C(control),D(leukocyte-depleted blood reperfusion),M(maerophage inhibition)and DM(leukocyte-depleted plus macropbage inhibition).The 20 cases of left lung transplantations were performed by the same surgeon.All procedures were identical,except that the donors in Group M and DM received the macrophage inhibitor gadolinium chloride(14 mg/kg)intravenously 24 h before operation,and that the recipients in Group D and DM underwent initial 10 min reperfusion with leukocyte-depleted blood collected from donors'inferior vena cava. All lung allografts were reperfused for 2 h.Results:Compared with Group D and C,macrophage inhibition ameliorated PO_2/FiO_2 and mean pulmonary arterial pressure(mPAP)consistently after 30 min reperfusion in Group M and DM;the parameters of lung reperfusion injury(malonaldehyde activity,wet/dry ratio)at 120 min after reperfusion were also significantly improved(P

AIM To study the mRNA expression of histamine H_1 receptor in hepatocarcinoma of rats. METHODS Dimethylamino-azobenzene (DAB) was used to induce hepatocarcinoma in rats. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was adopted to analyse the relative expression of histamine H_1 receptor. And the base sequence of its PCR product was detected. RESULTS The relative mRNA expression of histamine H_1 receptor was significantly decreased in hepatic carcinoma tissue, compared with that part far from cancer and control group (P

Objective To investigate the significance and effect of ultrasonic diagnosis on the autologous bone marrow mesenchymal stem cells (MSC) in angiogenesis. Methods Twenty-four New Zealand rabbits were divided into experiment group (12) and the control group (12). Then rabbit bone marrow MSCs from experiment group were isolated, caltured and marked with Brdu. After ischemic hind limb animal model on all rabbits was set up, autologous bone marrow MSCs were directly injected into the ischemic hind limb muscles in experiment group while same volume normal saline was used in the control group. Two weeks after the implantation of autologous bone marrow MSCs, 2D and color Doppler flow imaging (CDFI) detection were used in rabbit femoral artery of the two groups to observe the inner diameter of the blood vessel, the peak velocity and the acceleration time. The disposition of transplaned cells and the state of angiogenesis in ischemic muscles were assessed using immunofluorescence staining. Results The results of 2D and Doppler ultrasound detection showed the inner diameter of the blood vessel and the peak velocity of the blood current in experiment group obviously higher than that of the control group , and the acceleration time was obviously smaller than that of the control group P<0.01. The immunofluorescence staining showed there were transplanted cells existed in transplanted portion and state of angiogenesis was supurior obviously than that of the control. Conclusions Bone marrow MSCs had the effect to promote angiogenesis. Implantation of autologous bone marrow MSCs was a simple and efficient therapeutic method for the ischemia hind limb. Using high-frequency ultrasound to detect femoral artery may provide a practical and useful method to evaluate the effect on implanted autologous bone marrow mesenchymal stem cells.

BACKGROUND: Studies indicated that lipid peroxidation due to increase of free radical is the key factor of ischemia/reperfusion injury.Shinyleaf pricklyash root extracts, rutaceae plant, is bitter in taste, no stimulation, which has the effects of promoting qi, relieving pain, promoting blood circulation by removing blood stasis, dispelling wind and dredging collaterals and antioxidation.OBJECTIVE: To observe the influence of nitidine chloride on myocardial ischemia/reperfusion injury in rats and analyze its mechanism.DESIGN: Randomized controlled study.SETTING: Departmentof Pharmacology and Department of Chemistry,Guangxi Medical University.MATERIALS: A total of 60 healthy Wistar rats were selected, half male and half female, with the body mass of 250-300 g. Nitidine chloride was provided by Department of Chemistry, Guangxi Medical University, batch number 20050609. MS4000U biological signal quantitative record analysis system, 722N evident spectrophotometer, hydrochloric acid verapamil (batch number 020701, 2 mL in each), malondialdehyde (MDA) and superoxide dismutase (SOD) kit were purchased from Guangzhou Longfeida Technology Co., Ltd., Shanghai Precision Scientific Instruments Corporation, Shanghai Harvest Pharmaceutical Co, Ltd. and Nanjing Jiancheng Bioengineering Institute, respectively. Hitachi 7170A full automatic biochemistry analyzer was also applied.METHODS: The experiment was performed at the Department of Pharmacology and Department of Chemistry, Guangxi Medical University between June 2004 and May 2006. ①Totally 60 healthy Wistar rats with normal ECG (half male and half female) were randomly divided into 6 groups:sham operation group, model group, 2, 1, 0.5 mg/kg nitidine chloride groups, positive control group with 10 rats in each group. The rats in the sham operation group received threading without deligation, and 90 minutes later the experiment was accomplished. Other 50 rats received left anterior descending branch of coronary artery deligation, ischemia for 30 minutes reperfusion for 60 minutes. 2 mg/kg verapamil, 2,1,0.5 mg/kg, 5 mL/kgnitidine chloride, saline of the same volume were injected into femoral vein in rats of the positive control group, different doses nitidine chloride groups and model group, respectively 10 minutes before deligating left anterior descending branch of coronary artery. ②Monitoring was conducted successively with standard limb Ⅱ lead ECG when performing reperfusion. Type,incidence rate and duration of cardiac arrhythmia were recorded within 60minutes. Change of ST segment was also recorded after reperfusion for 15minutes and 60 minutes. ③At the end of experiment, serum myocardial enzymology indexes were measured wi th full automatic biochemistry analyzer.MDA content and SOD activity in myocardial tissues were examined with thiobarbituric acid(TBA) method and xanthine oxidase (XOD) method, respectively. ④Measurement data and enumeration data between two groups were compared with t test and x2 test. MAIN OUTCOME MEASURES: Occurrence of cardiac arrhythmia, ECG ST segment elevation, change of serum myocardial enzymology indexes, MDA content and SOD activity of myocardial tissues in rats of each group.RESULTS: A total of 60 rats were involved in the result analysis. ①Degree of cardiac arrhythmia and ECG ST segment elevation of rats: The emergency time of cardiac arrhythmia in 1 and 2 mg/kg nitidine chloride groups was significantly later than that in the model group (P ＜ 0.05,0.01). The duration of cardiac arrhythmia in the 1 and 2 mg/kg nitidine chloride groups and positive control group was obviously shorter than that in the model group (P ＜ 0.05-0.01). The incidence rates of various kinds of cardiac arrhythmia were markedly less than those in the model group (P ＜ 0.01). The degree of ST segment elevation at reperfusion for 15 and 60 minutes was remarkably lower than that in the model group (P ＜ 0.05-0.01). ②Serum myocardial enzyme level: It was significantly higher in the model than the sham operation group after 60-minute myocardial ischemia and reperfusion (P?.01). Activity of myocardial enzyme in the 1 and 2 mg/kg nitidine chloride groups was remarkably lower than that in the model group (P ＜ 0.01,P ＜ 0.05). The level of myocardial enzyme decreased with the increase of nitidine chloride. It was lower significantly in the positive control group than the model group (P ＜ 0.05-0.01 ). ③SOD activity of myocardial tissues: It was markedly lower in the model group than the sham operation group after 60-minute myocardialischemia and reperfusion (P ＜ 0.01); It was dramatically higher than in the 1 and 2 mg/kg nitidine chloride groups than the model group (P ＜ 0.01). The activity also increased with the increase of nitidine chloride. ④MDA content of myocardial tissues: It was distinctly higher in the model group than the sham operation group after myocardial ischemia reperftsion for 60 minutes (P ＜ 0.01). It was remarkably lower in the 1 and 2 mg/kg nitidine chloride groups than the model group (P ＜ 0.01). The content decreased with the increase of nitidine chloride. It was obviously lower in the positive control group than the model group (P ＜ 0.05 ).CONCLUSION: ①1 and 2 mg/kg nitidine chloride can reduce the incidence rate of cardiac arrhythmia in rats with myocardial ischemia and reperfusion, postpone the emergence time of cardiac arrhythmia and shorten its duration, decrease the degree of ST segment elevation after reperfusion for 15 minutes and 60 minutes, which have similar effect with verapamil.② 1 and 2 mg/kg nitidine chloride can reduce the release of myocardial enzyme, relieve the severity of oxygen-derived free radicals injury, and has the effect of protecting myocardial injury during ischemia-reperfusion, in which represents a dose-dependent effect.

Objective To establish a stable,repeatable and long-lasting rat model of myocardial infarction,and to evaluate the feasibility of monitoring electrophysiological changes and left ventricular function after myocardial infarction by electrocardiography (ECG) and ultrasonic cardiography(UCG). Methods Wistar rats were ligated on the left anterior descending coronary artery after anaesthesia with 10% chloral hydrate and machinery assisted respiration. Then they were monitored by ECG and UCG afer 4,8 and 12 weeks,and were sacrificed and pathologically examined at 12 weeks after operation. Results The rat model of myocardial infarction was established with a survival rate of 83.3% at 72 hours after the operation and 73.3% at 12 weeks after the operation. In the myocardial infarction group,the PR,QRS,QT and QTc intervals were statistically significantly longer than that in the sham operation group. UCG showed that the left ventricular internal diameter at end-diastole (LVIDd) and the left ventricular internal diameter at end-systole (LVIDs) were statiscally significantly higher in the infarction group,and the ejection fraction (EF) and fractional shortening (FS) significantly lower than those in the sham operation group. Long-lasting pathological changes can be seen in the tissues at 12 weeks after operation. Conclusions The method used in the present study is an simple,less injurious and highly successful technique,and the changes in electrophysiology and left ventricular function can be well monitored by ECG and UCG at different times during this period.

Objective To analyse MMACHC mutations for 45 pedigrees with combined methylmalonic aciduria and homocyctinuria by Sanger sequencing, and to discuss the utility of prenatal genetic diagnosis for these pedigrees.Method Peripheral blood was collected from 45 probands and their parents from 2012-2015 in Genetic Counselling Clinic of the First Affiliated Hospital of Zhengzhou University, and the DNA were extracted from the blood.Then the coding sequence of MMACHC gene was amplified by PCR, and the PCR products were further sequenced to detect mutations for each pedigree.For 12 families, chorionic villus sampling was performed on the pregnant women to make prenatal genetic diagnosis.Result There were 14 distinct mutations detected in the 45 pedigrees, and the most frequent mutations are c.609G>A(W203X),c.658-660delAAG(K220del)and c.80A>G (Q27A).Two of those mutations have not been reported before:one is a splicing site mutation c.81+1G>A;while the other is a missense mutation c.665A>G,p.Y222C.Most mutations were found in exon 4.Among the 12 pedigrees who received prenatal diagnosis, 2 fetuses were normal, 7 fetuses were carriers of heterozygous mutation, and the other 3 fetuses were patients with compound heterozygous mutation or homozygous mutation.The couples whose fetuses were normal or carriers continued the gestation, while the couples whose fetuses were patients decided to terminate the pregnancy.After delivery, the outcome of the fetuses was the same as the prenatal diagnose results.Conclusion Two novel mutations of MMACHC were identified and prenatal genetic diagnosis helps to avoid the delivery of combined methylmalonic aciduria and homocyctinuria patients.

Objective To analyze the mutation of MUT with Sanger sequencing technology to explore the feasibility of its application in prenatal diagnosis.Methods MUT sequencing was performed in 24 pedigrees who had history of isolated methylmalonic acidemia (MMA) babies and came to the First Affiliated Hospital of Zhengzhou University and Newborn Screening Center of Maternal and Child Health Hospital of He'nan Province between October 2012 and June 2015 for genetic counseling.Meanwhile,another 100 cases of normal controls also had their MUT gene sequence analyzed.After confirming the genotype of each pedigree,we collected the villi of nine high-risk fetuses in nine pedigrees whose parents were prepared for prenatal diagnosis.Results Totally,25 kinds of MUT gene mutations were identified among the 24 isolated MMA pedigrees,in which 11 were novel mutations including one nonsense mutation [c.616C＞T(p.Q206X)],six missense mutations [c.613G＞A(p.E205K),c.894T＞G(p.1298N),c.1009T＞C(p.F337L),c.1154G＞T(p.L385W),c.1663G＞A(p.A555T) and c.1675G＞A(p.R559G) and four frame shift mutations [c.626-627insC(p.P209Pfs*2),c.755-756insA(p.H252Qfs*6),c.756-757insA(p.M253Nfs*5) and c.1581-1582insA(p.A528Ifs*4)].None of the above mutations was detected in the controls.Finally,among the nine pedigrees for prenatal diagnosis,two were determined to have normal MUT gene,four were found to be heterozygous mutation carriers of MUT gene and three were confirmed as complex heterozygous or homozygous mutation carriers.Families of fetus who had normal MUT gene or fetuses who were carriers chose to continue the pregnancy,while those who had heterozygous mutation of MUT gene chose termination.The results of follow-up of newborns were consistent with that of prenatal diagnosis.Conclusions We found two novel mutations in MUT gene that might lead to isolated MMA.And Sanger sequencing technology for MUT gene sequencing analysis might effectively avoid the birth of isolated MMA children.