Hepatitis B virus exists in the form of quasispecies.Quasispecies are an important basis for the pathogenesis and drug resistance of hepatitis B virus.Researches for quasispecies could contribute to evaluating the progress of disease, predicting drug resistance, adjusting the therapeutic regimen and performing individual treatment.

Objective To reseach the time point of the highest percentage of neural precursor cells derived from adipose stromal cells (ADSCs) in vitro, and to observe the ultrastructure features of neural precursor cells. Methods Used the β-mercaptoethanol to induce ADSCs to differentiate into neural precursor cells and neuron-like cells. The morphology of the uninductedcells and inducted cells were observed with inverted phase contrast microscope. The expression of nestin which was the marker of neural precursor cell in each group was detected using immunofluorescence staining method. The ultrastructural feature of cells which was induced for 3 hours were observed. Results The highest ratio of positive expression of nestin was 3 hours following induction,with the ratio ( 86.25 ± 4.82) %. There were many protuberance on the cell membrane under transmission electron microscopy.There were plenty of organelles in the neural precursor cells. The neural precursor cells had a large size nucleus,large nucleoplasmic index, much extended chromatin,and less condensed chromatin. The nucleus had double-layer nuclear envelope, more nuclear pore on the nuclear envelope. Conclusion The time point of the highest percentage of neural precursor cells derived from ADSCs is 3 hours,and the ultrastructral feature of induced neural precursor cells confirm that cells at this time point are in a state of split active period.

Objective To induce adult adult adipose-derived stem cells(ADSCs) in vitro to differentiate into neuronal-like cells,and to analyze the features of their cell morphology and ultrastructure. Methods Adipose stromal cells were obtained and amplified in vitro. Then make use of chemical induction to induce them. Observed ADSC and differentiation of cells in morphology and ultrastructure under inverted microscope and transmission electron microscopy. Immunocytochemistry to detection of Nestin, Neuron Specific Endolase( NSE) ,and glial fibrillary acidic protein(GFAP) expression in cells. Used Real-time PCR to detection of Nestin,GFAP gene mRNA expression before and after induction in ADSC. To observe the morphology and ultrastructure of the cells prior to and after induction under microscope and electron microscope. Results The morphology of ADSC was similar to fibroblasts ,and could be amplified stability within 10 passages in vitro. Some of the cells induced display a typical astrocyte-like cells in ultrastructure. Followed neuronal induction,astrocyte-like cells began to stain brightly for CFAP, Nestin. GFAP stained in both the cytoplasm and nuclei of astrocyte-like cells, but Nestin only stained in the cytoplasm. The peak positive expression rate within 14d following neuronal induction. The rate of positive expression cells was( 14.4 ± 3. 6) % for Nestin, (87. 3 ± 5. 3 ) % for GFAP. Then two kinds of protein expression remained the similar rate. The average relative concentration of GFAP and Nestin gene mRNA have significant statistical difference between ADSC and differented cells analyzed by Real-time PCR (P<0.05).The peak concentration of GFAP was within 20 d after induction,and GFAP was within 14 d after induction. Conclusion In the cytoplasm of adult adipose-derived cells possess Nestin genetic material,which is the marker of neural stem cell. The differential astrocyte-like cells have the typical morphology, ultrastructure and GFAP phenotype of mature astrocytes.

Objective To retrospectively investigate the clinical features and incidence trends of hypertriglyceridemic acute pancreatitis (HLAP) in Guangzhou First People's Hospital for 20 years.Methods The medical records of 1 362 patients,who were admitted to Guangzhou First People's Hospital with acute pancreatitis during January 1991 to December 2010,were reviewed and 99 patients met the HLAP criterion.These patients were divided into four groups according to years (1991 ～ 1995,1996 ～2000,2001 ～2005,and 2006 ～2010).The incidence and clinical features were compared among the 4 groups.Results Among the 99 patients,there were 61 males and 38 females,the mean age was (44 ± 12) years old,and young male was predominant in HLAP.HLAP accounted for 7.3％ of all AP patients,and the incidence was increased 1.5 times during the 20 years period (5.4％,5.7％,6.7％,and 8.3％ respectively).The serum amylase level was (513.3 ±462.7) mmol/L,TG level was (12.7 ± 7.0) mmol/L,Ranson score was (1.2 ± 1.1),CTSI score was 2.2 ± 1.1,incidence of SIRS was 36.4％,and the incidence of MODS was 18.2％,and the incidence of pancreatic pseudocyst was 5.1％,but the incidence and mortality of SAP was not changed with time.The incidence and mortality of SAP of HLAP were significantly higher than those of acute biliary pancreatitis (20.2％ vs 8.5％,6.1％ vs 2.8％).The rate of blood purification increased from 0 to 10.7％,and the hospital stay was decreased from 26 d to 14 d,and the difference between the two groups was statistically significant (P ＜ 0.05).Conclusions Incidence of HLAP is increased gradually,with a trend towards more severe disease,but the prognosis is improved.

OBJECTIVETo evaluate the effect of Shexiang Baoxin Pill (SBP) on coronary vasodilation by analysis of coronary angiography (CAG).

METHODSA consecutive cohort of 300 patients who underwent CAG between January 2013 and July 2013 were recruited and randomly assigned to 2 groups before operation. Patients in the SBP group sublingually took SBP, while those in the control group sublingually took placebos. All patients repeatedly underwent CAG 5 min after administration. The vascular diameter was calculated by quantitative angiography analysis method. The diameter of the left anterior descending coronary artery was measured in patients whose coronary arteries had no stenosis. The narrowest vascular diameter was measured in patients whose coronary arteries had stenosis. The heart rate, blood pressure, and the vascular diameter were compared between before and after administration in the two groups.

RESULTSIn the two groups, there was no significant difference in changes of heart rate, systolic pressure, or diastolic pressure between before and after administration (all P > 0.05). There were 64 patients with normal CAG in the two groups, 30 in the control group and 34 in the SBP group. CAG showed there were 236 patients with stenotic coronary artery, 110 in the control group and 126 in the SBP group. The vascular diameter was obviously larger in patients in the SBP group with normal or abnormal CAG after administration (all P < 0.01). It was also obviously larger than that of the control group after administration (P < 0.05, P < 0.01).

CONCLUSIONSBP could dilate both normal coronary artery and lesioned coronary arteries, but did not lead to fastened heart rate and decreased blood pressure.

Blood Pressure ; Coronary Angiography ; Coronary Vessels ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Rate ; Humans ; Tablets ; Vasodilation ; drug effects

OBJECTIVE: To choose a PPK model of vancomycin most suitable for Chinese pediatric patients, in order to guide the dosage adjustment. METHODS: Based on the database of PubMed and CNKI, all studies regarding vancomycin population pharmacokinetics were investigated and their basic information including PPK models was extracted. The data of Chinese pediatric patients who were administered vancomycin and received therapeutic drug monitoring (TDM) were introduced into the reported final PPK models, and the fitting was conducted by model fitting graphics. RESULTS: Twelve vancomycin PPK studies during 1986-2014 were included. As judged by the correlation coefficient (R) and R-square (R2) between predicted concentration and measured concentration, the models 4 and 9 presented a relatively better fitting with the data of Chinese pediatric patients we collected. The VPC fitting demonstrated that number 1 model achieved the best fitting. However, because the limited data used in this study was based on irregular sampling time, so the VPC test results were difficult to be distinguished and could only be used as a secondary reference. On the other hand, NPDE has corresponding statistical test, and its evaluation ability for the model is not affected by the factors of the experimental design. NPDE analysis showed that one-compartment model was better than two-compartment model, and model 4 and model 9 achieved better fitting to the collected data than others. CONCLUSION: The fitting effects of most reported vancomycin PPK models, except individual models, were poor for the TDM data of Chinese pediatric patients, therefore it is necessary to establish a vancomycin population pharmacokinetic model particularity for Chinese pediatric patients, in order to guide dosage adjustment more accurately.

OBJECTIVETo observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms.

METHODSWe randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication.

RESULTSThe total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C.

CONCLUSIONShugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.

Asthenozoospermia ; drug therapy ; enzymology ; Biomedical Research ; Capsules ; Carnitine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Semen ; enzymology ; Spermatozoa ; alpha-Glucosidases ; analysis

Objective To evaluate the effects of infliximab (TNF-α monoclonal antibody ) on intestinal barrier injury in ANP complicated with MODS in a rat model.MethodsThirty SD rats were randomly divided into sham operation group (SO),ANP group and infliximab treatment group.Sodium taurocholate (4.5％) was injected into the pancreatic duct to induce ANP complicated with MODS model.Infliximab (8 mg/kg) was injected via tail vein in 6h after modeling in infliximab group.Same amount of 0.9％ NS was injected into the pancreatic duct in SO group.After 24 h of modeling,all rats were sacrificed,intestine and pancreas samples were collected for pathologic examination.The blood samples were harvested.The serum levels of amylase,TNF-α,diamine oxidase( DAO),D-lactate,and the rate of carbon propelling in ileum were measured.ResultsThe serum levels of amylase were ( 1125 ± 331 ),( 11024 ± 2203 ),( 545 ±30) U/L in SO group,ANP group and infliximab group; the serum levels of TNF-α were (12.1 ± 4.0),(107.6 ± 18.5),(75.8 ±5.9) U/L; the pathological scores of pancreas were 2.25 ±0.38,14.10 ±0.22,3.93 ± 0.67,the difference among the 3 groups was statistically significant ( P ＜ 0.05 ).The pathological scores of intestine were 2.29 ± 0.32,6.61 ± 0.58,3.91 ± 0.41 ; the DAO levels were ( 87.88 ± 34.51 ),(146.30 ±12.99),(115.00 ± 18.58) ng/ml; the D-lactate levels were (1.50 ±0.49),(2.32 ± 0.35),(2.02 ± 0.25 )mmol/L; and the rates of carbon propelling in ileum were (0.64 ± 0.04 )％,(0.28 ±0.08)％,(0.52 ±0.09)％,the difference among the 3 groups was statistically significant (P ＜0.05).ConclusionsInfliximab can effectively prevent dysfunction of intestinal barrier and improve motility in ANP rats.

Objective To observe the treatment of ~(99)Te-MDP on typeⅡcollagen induced arthritis (CIA)in rat,and the effect on the expression of synovial MMP-3 and TIMP-1 mRNA.To explore the mech anisms of the ~(99)Te-MDP in the treatment of rheumatoid arthritis.Methods The rats in which C1A(n=24)were divided into three group:the control group(n=8),~(99)Tc-MDP group(n=8)and Methotrexate group(n=8). Arthritis were evaluated by arthritis index and histopathological index and the expressions of MMP-3 and TIMP-1 mRNA in synovium were detected by RT-PCR.Results①The arthritis indexs of the control group, the methotrexate group,the ~(99)Tc-MDP group were increased with time.②The histopathological scnres of the control group were significantly higher than those of methotrexate group and ~(99)Tc-MDP group(P＜0.01).The histopathological scores of cartilage destruction and bone erosion of ~(99)Tc-MDP group were lower than those of methotrexate group(P＜0.05).③The levels of MMP-3 mRNA of the control group,~(99)Tc-MDP group, methotrexate group were notably higher than those of the control group(P＜0.01).The levels of control group was notably higher than that of the ~(99)Tc-MDP group(P＜0.05).There was not significant difference in all groups on the levels of TIMP-1 mRNA(P＞0.05).Conclusions ~(99)Tc-MDP can notably relieve the arthritis symdrome and retard the catilage damage and bone erosion of CIA in rats,and could significantly decrease the MMP-3 mRNA in the synovium.Which may be one of the therapeutic mechanism.~(99)Tc-MDP is better than methotrexate in retarding catilage and bone erosion and decreasing MMP-3 mRNA in CIA rats in a 3-week therapeutic intervention.

Objective To demonstrate high mobility group box chromosomal protein 1(HMGBI) expression in synovium and joint,and to identify the role of HMGB1 in the pathogenesis of synovitis and joint destruction in adjuvant-induced arthritis(AA).Methods AA of 15 male rats were induced in SD rats by intradermal injection of 100?l Freud's complete adjuvant in the foot pad of the left hind paw.All rats were killed at the 18th day.Synovium and joints were collected for histopathology studies and determining the expression of HMGB1 by immunohistochemistry,and serum was collected for determining the expression of HMGB1 by western blotting analysis.Results Immunostaining of specimens from normal rats showed that HMGB1 was primarily confined to the nucleus of synoviocytes with occasional cytoplasmic staining.In contrast, inflammatory synovial tissues from AA rats showed a distinctly different HMGB1 staining pattern.Nuclear HMGBI expression was accompanied by a cytoplasmic staining in many mononuclear cells.The cytoplasmic HMGB1 expression in synovium of AA rats is significantly higher than that of normal rats.Additionally,HMGBI was highly expressed in the nuclei and cytoplasm of the subchondral chondrocytes and inflammatory cells in bone erosion in AA rats(P＜0.01),while fewer positive cytoplasmic staining of HMGB1 was found in chondrocytes and fewer positive nuclear staining was found in bone cells in normal rats.HMGB1 concentration was significantly higher in serum of AA rats than that in normal rats(P＜0.001).Conclusion The cytoplasmic HMGBI expression in synovium and joints is greatly upregulated;the level of HMGB1 in serum is increased in AA rats which suggests a patbogenetic role of HMGB1 in synovitis and bone destruction of adjuvant-induced arthritis.