Aim To establish a HPLC method for determination of the content of Huperzine A in Huperzia serrata and compare it in different parts of the plants. Method DiamonsilTm C_ 18 column(200 mm?4.6 mm,5 ?m)was used; the mobile phase was methanol ∶0.08 mol?L-1ammonium acetate buffer(40 ∶60,pH=6.0),the detection wavelength was 308 nm, with flow rate at 1.0 ml?min-1;the column temperature was room temperature.Results There is a good linear relationship within the range of 5～100 mg?L-1(r=0.9997).The average recovery was 99.10%,with RSD 2.44%(n=5).Regarding the content of Huperzine A in different parts of Huperzia serrata, it was found that the quantity of Huperzine A in leaf was higher than that in stem and it was the lowest in root. Conclusion The method is simple, accurate and reliable etition. Huperzine A can be accumulated in the aboveground part of the plant.

AIM: To evaluate the quality of Rhizoma gastrodiea processed by different drying methods. METHODS: Vacuum freeze drying and oven drying were adopted. The content of gastrodin was determined by RP-HPLC. RESULTS: The content of gastrodin of Rhizoma Gastrodiae processed by vacuum freeze drying was significantly higher than that by boil with water and oven drying. CONCLUSION: The method of vacuum freeze (drying) can prevent Rhizoma Gastrodiae from degradation of gastrodin, to keep the appearance, to make Rhizoma Gastrodiae convenient for further processing.

Stroke is responsible for increasingly high rates of mortality and disability worldwide. Approximately two million people suffer from stroke for the first time in China each year. The high incidence (50%) of post-stroke disability brings a heavy burden to patients and their caregivers. Acupuncture has been widely used in the communities for post-stroke rehabilitation in China. The objective of this trial is to apply our acupuncture research achievement to treatment and evaluation of post-stroke hemiplegic patients in community.

A cidithiobacillus ferrooxidans is a chemolithotrophic microorganism capable of using ferrous ions and sulphides as energy sources. This microorganism has an important role in the bioleaching of minerals. During this process, the bacteria are normally subjected to several stressing conditions, such as temperature changes, lack of nutrients or pH changes, which may affect the efficiency of the bacterial action. SELDI is a recent technology that allows for high-throughput proteomics studies. The Protein Chip SELDI technology was used to generate comparative protein profiles of A c idithiobacillus ferrooxidans grown under phosphate starvation or normal condition additionally adding Fe2+ as energy resource. There were 13 significantly differential expressed protein's peaks found by using SELDI Protein Chip technologies, which made a solid foundation for further isolation these low molecular proteins by adopting technologies such as HPLC etc.

Objective To study the mechanism of restenosis following percutaneous transluminal coronary angioplasty(PTCA),and to replicate a dynamic model of cell proliferation and remoulding of vascular wall at different time points in rabbits after intimal injury.Methods The model of restenosis in common carotid artery was established by balloon injury in 70 rabbits.The indexes such as lumen area,thickness and area of intima and media,and cross sectional area bounded by the external elastic lamina(EELA) were respectively measured by computer image analysis technology at the 1st,3rd,5th, 7th,14th,28th and 35th day after the injury.Results Endothelial cells were denudated at the 1st day after injury.The proliferation of vascular smooth muscle cell(VSMC) was detected on the surface of lumen at 3 days after injury.At the 7th day after injury,the neointima was formed and continuously thicken.The thickness and area of the neointima as well as extracellular matrix were gradually increased after 14 days,and were maximal after 35 days.The thickness and area of media were also gradually increased during 3～14 days and decreased after 28 days.Compared with non-injured vessel,the medial area was obviously increased at the 14th day.The lumen area was decreased at the 5th～7th day after injury and was obviously less than that of non~injured vessel after 14 days.The EELA was gradually increased at the 1st～7th day after injury,and reached its maximum at the 14th day.The EELA was declined gradually after 28 days.Conclusion The progress of restenosis(RS) can be simulated through the model of restenosis in common carotid artery of rabbit established by balloon injury.The intimal proliferation and vascular remodeling are the leading pathogenesis of restenosis.

Objective To compare the clinical effect of two kinds of internal fixation of locking plate and anatomical plate in the treatment of Pilon fracture.Methods 64 patients with Pilon fracture undergoing elective surgery were randomly divided into study group and control group by digital table method,32 cases in each group.The study group received locking plate internal fixation for the treatment,the control group was treated with anatomical plate internal fixation.The clinical efficacy and safety of two kinds of internal fixation methods were compared.Results After treatment,the excellent rate of ankle joint function in the study group(96.88%)was significantly higher than that in the control group (81.25%),and the difference had statistical ssignificance (χ2 =4.01,P <0.05 ).The operative time,hospitalization time and fracture healing time[(57.32 ±10.98)min,(13.72 ±591 )d,(15.72 ± 2.82)weeks]in the study group were shorter than those in the control group[(68.11 ±11.85 )min,(16.62 ± 5.21)d,(17.81 ±3.42)weeks],and the differences had statistical significance(t =3.98,3.11,321,all P <0.05). The amount of bleeding (153.32 ±21.36)mL in the study group was less than that in the control group [(188.65 ± 26.24)mL](t =5.46,P <0.05).The incidence rate of delayed fracture healing of the study group (3.13%)was significantly lower than that in the control group(28.13%),the difference had statistical significance(χ2 =7.59,P <0.05).There were no significant differences in the incidence of other complications between the two groups(χ2 =1.95,102,102,0.35,all P >0.05).Conclusion Locking plate fixation in the treatment of Pilon fractures could significantly promote the recovery of joint function,and has the advantages of short operation time,fast fracture healing,less complications and other advantages,it is better than anatomical plate internal fixation.

Objective To evaluate the long-term effect of surgical procedures for congenital choledochal cyst (CCC).Methods From 1986 to 2000, 120 cases of CCC were admitted and 73 of them underwent the primary operations in General Hospital of PLA. Three types procedures were performed,type I: external drainage of CCC in 7 cases; type II:cystojejunal Roux-en-Y anastomosis in 5 cases; type III: cyst excision with cystojejunal Roux-en-Y anastomosis or cystoduodenostomy in 57cases,and other procedures in 4 cases.Results 68 cases were followed-up for 6 months to 5 years (median 2.7 years). Three cases undergoing type I operations accepted reoperations;two cases undergoing type II operations accepted reoperations due to severe complications as cholongitis and hepatolithiasis; 57 cases treated by type III operation with the good results 88.7% and none reoperation.Conclusions External drainage is only a first-aid management on emergency basis. Internal drainage should never be done,because the effect is temporary,and severe complications result in reoperations. Cyst excision with biliary tract reconstruction is recommended as the optimal treatment of CCC.

Objective:To explore the correlation of the expression of lymphocyte immunoglobulin-mucin domain 3 (Tim-3) on T lymphocytes and natural killer (NK) cells with hepatic inflammation and hepatic fibrosis in patients with chronic hepatitis B virus (HBV) infection.Methods:A total of 320 patients of chronic HBV infection who visited the Infectious Diseases Department in the Third Affiliated Hospital of Sun Yat-sen University from June 2016 to June 2018 were enrolled. The patients were divided into four groups: immune tolerant group (IT, n=31), immune active group (IA, n=184), inactive carriers group (IC, n=48), and gray zone group (GZ, n=57). And 17 healthy controls (HC group) were included at the same time. Peripheral blood mononuclear cells were separated and the frequency and mean fluorescence intensity (MFI) of Tim-3 on T cells (CD3 + , CD4 + and CD8 + T cells) and NK cells (NK, NK-bright and NK-dim cells) were detected by flow cytometry. The clinical data of patients were collected and aspartate aminotransferase-to-platelet ratio index (APRI) score was calculated. Kruskal-Wallis H test was used for comparing the data of non-normal distribution among groups, and Mann Whitney U test was used for the comparison between two groups. Enumeration data were expressed as cases (percentage) and compared by the Chi-square test. Spearman rank correlation was used for correlation analysis. Receiver operating characteristic curve (ROC) was used to analyze the predictive value of Tim-3 expression on T cells and NK cells in evaluating liver fibrosis in patients with chronic HBV infection. P<0.05 was considered statistically significant. Results:Significant differences were found in the age, aspartate aminotransferase (AST), alanine aminotransferase(ALT), albumin (Alb), total bilirubin (TBil) and liver stiffness measurement (LSM) among IT, IA, IC, GZ and HC groups ( H=12.40, 169.70, 210.70, 25.17, 24.21 and 86.5, all P<0.05). And the differences in APRI score, proportion of HBeAg-positive patients, HBsAg and HBV-DNA among the IT group, IA group, IC group, GZ group were also significant ( H=89.45, 118.00 and 14.81, χ2=148.20, all P<0.05). The frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells, NK cells, NK-bright and NK-dim cells among the IT group, IA group, IC group, GZ group and the HC group were significantly different( H=13.57, 51.55, 8.58, 44.25, 20.32, 47.96 and 12.45, 33.69, 4.96, 32.47, 10.63, 30.46, all P<0.05). Both of the frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells were positively correlated with ALT and AST levels in patients with chronic HBV infection ( r=0.2134, 0.4733, 0.2090, 0.4333, 0.1771, 0.4417, 0.1780, 0.3956, 0.2618, 0.4671, 0.2614 and 0.4326, all P<0.05). While the frequency and MFI of Tim-3 on CD8 + T cells and MFI on CD3 + and CD4 + T cells were also positively correlated with TBil levels ( r=0.1342, 0.2635, 0.2739 and 0.2526, all P< 0.05). The frequency and MFI of Tim-3 on NK and NK-dim cells were negatively correlated with the levels of ALT, AST and TBil ( r=-0.2671, -0.4093, -0.2451, -0.4099, -0.1807, -0.1823, -0.2733, -0.4224, -0.2576, -0.4206, -0.1798 and -0.1946, all P<0.05). The MFI of Tim-3 on NK-bright cells was also negatively correlated with ALT, AST and TBil ( r=-0.3775, -0.3562 and -0.1633, all P<0.05). Both of the frequency and MFI of Tim-3 on CD3 + , CD4 + and CD8 + T cells were positively correlated with liver fibrosis( r=0.1789, 0.3896, 0.1518, 0.3521, 0.2117 and 0.3579, all P<0.05). Both of the frequency and MFI of Tim-3 on CD4 + and CD8 + T cells and the MFI of Tim-3 on CD3 + T cells were positively correlated with APRI score ( r=0.1487, 0.2604, 0.2296, 0.4858 and 0.2853, all P<0.05). The expression frequency and MFI of Tim-3 on NK and NK-dim cells and MFI of Tim-3 on NK-bright cells were negatively correlated with LSM ( r=-0.2686, -0.3975, -0.2852, -0.3991 and -0.3531, all P<0.05). The expression frequency and MFI of Tim-3 on NK and NK-dim cells and MFI of Tim-3 on NK-bright were negatively correlated with APRI score ( r=-0.3589, -0.4158, -0.3591, -0.4108 and -0.3966, all P<0.05). The ratio of Tim-3 expression on CD3 + T cells to that on NK cells was shown to be able to predict liver fibrosis in chronic HBV infected patients and the area under the ROC curve was 0.783 (95% CI: 0.723~0.843, P< 0.05), and when the cut-off value was 0.612, the sensitivity was 61.9%, and the specificity was 99.3%. Conclusion:The relationship of Tim-3 expression on T cells with liver inflammation and fibrosis is opposite to that on NK cells in patients with chronic HBV infection, indicating that the ratio of Tim-3 expression on T cells to that on NK cells may be valuable in evaluating liver fibrosis in patients.

Objective: To analyze the changes and significance of serum soluble programmed death-1 (sPD-1) in patients with chronic hepatitis C (CHC) and study its role in the progression of CHC. Methods: Serum levels of sPD-1 in CHC patients and healthy controls (HC) were measured using ELISA and compared. Correlations of serum sPD-1 with peripheral hepatitis C virus (HCV) RNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver fibrosis (indicated by APRI) were analyzed. Results: The serum sPD-1 level in the CHC group was significantly higher than that in the HC group (P<0.05), and positively correlated with peripheral HCV RNA, ALT and AST (P<0.05). In addition, the serum sPD-1 levels in patients with APRI greater than 1.2 (indicating severe liver fibrosis) were higher than those in patients without or with mild liver fibrosis (P<0.05). Conclusions sPD-1 might be involve in the progression of CHC. Measuring serum sPD-1 in CHC patients would assist the prediction of disease progression and help to make the correct diagnosis and appropriate clinical therapy decision.