Objective: To prepare rabbit polyclonal antibody against human P-element-induced wimpy testis like 2 (HIWI2) protein, identify its properties and investigate its distribution in normal and tumor tissues by means of tissue chip. Methods: PIWIL2 peptide was synthesized chemically and conjugated to Keyhole limpet hemocyanin (KLH) as an immunogen. Then the PIWIL2-KLH conjugation was injected into rabbits subcutaneously to produce polyclonal antibodies. The specificity and sensitivity of antibodies were identified by ELISA and Western blotting after purification by affinity chromatography. PIWIL2 was then immuno-stained on the tissue chip to study its distribution in the normal and tumor tissues. Results: Rabbit's antibodies against human PIWIL2 were prepared after the injection of PIWIL2-KLH conjugation subcutaneously. These antibodies were identified as PIWIL2 peptides by ELISA and Western blotting assay. PIWIL2 protein expression was tissue-specific in tumor tissues, with PIWIL2 protein found in the cytoplasm of smooth muscle cells of most normal and tumor tissues. Conclusion: The polyclonal antibodies against human PIWIL2 protein have been successfully prepared, which provides a basis for further study on the role of PIWIL2 in the pathway of miRNA/RNA.

Objective To investigate the significance of MRI T2 fluid-attenuated inversion recovery hyperintense vessel sign (FLAIR HVS)in clinical prognosis evaluation of the patients with acute middle cerebral artery irnfarction.Methods The data in 57 inpatients with acute middle cerebral artery infarction in our hospital from Aug.2013 to Aug.2015 were retrospectively analyzed.All cases were performed the intact MRI examination(ineluding FLAIR,DWI and MRA)and CTA.The infarct volume with DWI,national institute of health stroke scale(NIHSS)score and modified Rankin Scale(mRS)score on 30 d after discharge were performed the comparative analysis.Results Fifty-seven cases of middle cerebral artery occlusion were divided into the distal HVSgroup and non-distal HVS group(8 cases in proximal HVS group,21 cases in HVS negative group).The infarction volume of DWI sequence,NIHSS scores at admissiom and discharge and mRS score on 30 d after discharge in the distal HVS group were superior to those in the non-distal HVS group(P＜0.05).Conclusion MRI-T2 FLAIR sequence HVS has certain reference value in the prognosis evaluation in the patients with middle cerebral artery occlusion.

The article made a study on the earliest literature and the initial time of the "Five Orbiculi" theory in TCM. By analyzing the masons for different locations of "Five Orbiculi" recorded in ancient TCM books, the author argued against that the term of "Five Orbiculi" originated from ancient India and the "Five Orbiculi" theory was a product with the combination of traditioinal Chinese and Indian cultures. The author further put forward that the "Five Orbiculi" theory most probably was a Chinese traditional medical innovation under the influence of Internal Classic of Medicine, a great development to the theory recorded in Miraculous Pivot On Serious Confusion.

Objective To investigate the best digestion time for rabbit rib cartilage cells separation by collagenase Ⅱ diges‐tion method .Methods Rabbit cartilage tissue was taken from 3‐month‐old New Zealand white rabbits ,and then digested in 0 .2%collagenase Ⅱ solution for 4 ,8 ,12 ,16 and 20 h under vibration at 37 ℃ ,respectively .The corresponding experiments were named as group A ,B ,C ,D ,and E .The obtained primary cells were cultured and passaged after counting .The cell morphology ,proliferation rate ,secretion of extracellular matrix of passage 2 cells were compared to chose the best digestion time .Results The primary chon‐drocytes obtained in group C ,D and E were more than the other two group A and B .The chondrocytes cytoactive in group C and D were better than the other three group A ,B and E .The passage 2 chondrocytes in group C ,D had better secretion of extracellular matrix capability than the other three groups .Conclusion The best collagenase Ⅱ digestion time is 12 to 16 h ,at this digestion time period ,more primary chondrocytes with high cytoactive and strong proliferation capability can be obtained ,and the phenotype of their passage 2 cells is quite good .

Objective To study the genomic molecular organization and genotype of human astro-virus infected infants in Shanghai of China. Methods Based on the published genomic sequence of HAstV (GenBank), the whole genome of one isolate human astrovirus was sequenced by specific primers. The PCR-products were cloned to pMD18-T vector and sequenced, phylogenetic tree was constructed by the Neighbor-Joining method with MEGA 4 software. Results The genome of HAstV-SH is 6807 hp, contains three ORFs: ORF1a and 1b encode the non-structural protein (from 83 nt to 4372 nt), ORF2 encodes the structural protein (from 4364 nt to 6727 nt). Compared with the ORF2 gene of those eight astrovirus sero-types in GenBank, revealed that the highest homology is with genotype 1 (97%). Homology with other gen-otypes ranged between 63% and 70%. Conclusion HAstV-SH belonged to genotype 1 and closely clus-tered with a strain of Japan (AB009985).

Objective To investigate the accuracy of serum S-100β protein and neuron specific enolase (NSE) level in predication of postoperative delirium (PD) in patients of different ages.Methods Four hundred ASA Ⅰ -Ⅳ patients of both sexes weighing 40-82 kg undergoing abdominal surgery performed under general anesthesia were divided into 4 age groups:group Ⅰ 18-44 yr; group Ⅱ 45-59 yr; group Ⅲ 60-74 yr and group Ⅳ ≥75 yr.The diagnosis of PD was made by using confusion assessment method.The incidence of PD was recorded within 72 h after operation.Each group was further divided into PD and non-PD subgroups.Blood samples were taken at 1 day before operation (T1),during their stay in PACU (T2) and at 24 and 72 h after operation (T3,4 ) for determination of serum S-100β protein and NSE concentrations.The receiver operating characteristic (ROC) curve for serum S-100β protein concentration in determining the PD efficacy was plotted.Results The incidence of PD was significantly higher and the duration was significantly longer in groups Ⅲ and Ⅳ than in groups Ⅰ and Ⅱ,and in group Ⅳ than in group Ⅲ (P ＜ 0.05).There was no significant difference in the serum S-100β protein concentration between PD subgroup and non-PD subgroup in groups Ⅰ - Ⅲ ( P ＞ 0.05).Compared with that at T1 and in nonPD subgroup,the serum S-100β protein concentration was significantly increased in PD subgroup in group Ⅳ,and the serum NSE concentration was significantly decreased at T2,3 in PD subgroup in group Ⅰ (P ＜ 0.05).There was no significant difference in the serum NSE concentration between PD subgroup and non-PD subgroup in groups Ⅱ -Ⅳ.The analysis results of the ROC curve showed that:the area under the curve for the serum S-100β protein concentration and 95％ confidence interval were 0.329 (0.127-0.531),0.352 (0.168-0.536),0.619 (0.466- 0.772) and 0.921 (0.846-0.995),the sensitivity was 50％,50％,56％ and 88％,and the specificity was 29％,22％,46％ and 86％ in groups Ⅰ-Ⅳ respectively.Conclusion Increase in the serum S-100β protein concentration can be used in predicting the development of PD in patients ≥75 yr,but the serum NSE protein concentration can not be used.

ObjectiveTo investigate the effect of recombinant interleukin-2 (rIL-2) combined with allicin in the treatment of pancreatic cancer and related mechanisms. MethodsA nude mouse xenograft model was established. A total of 60 nude mice were randomized into control group, rIL-2 treatment group, allicin treatment group, and combined treatment group. At 4 weeks after treatment, peripheral blood was collected, tumor volume, tumor weight, and survival time were recorded, and survival rates were calculated. Flow cytometry was used to analyze the apoptosis of tumor cells and measure the percentages of CD4+ T, CD8+ T, and natural killer (NK) cells, ELISA was used to measure the level of interferonγ (IFNγ), and Western blot was used to measure the expression of Bcl-2 protein in tumor tissue. An analysis of variance was used for comparison of continuous data between groups, and SNK-q test was used for further comparison between any two groups. ResultsAt 4 weeks after treatment, the rIL-2 treatment group, allicin treatment group, and combined treatment group showed significant reductions in tumor volume compared with the control group (all P＜0.05), and the combined treatment group showed the greatest reduction (P＜0.01). The combined treatment group had a tumor inhibition rate of 90.5% and a prolonged survival time (60% of the mice survived at 55 days). Compared with the control group, the combined treatment group showed significant increases in the apoptosis rate of tumor cells (233%±4.3% vs 90%±3.7%, P＜0.05) and the expression of pro-apoptotic protein Bcl-2. The treatment groups showed significant increases in the percentages of CD4+ T, CD8+ T, and NK cells compared with the control group (F=23.74, 26.38, and 1972, all P＜0.001). Compared with the other three groups, the combined treatment group showed a significant increase in the IFNγ level (F=9.84, P=0.026). ConclusionCombined treatment with allicin and rIL-2 can enhance the innate immunity and adaptive immunity and thus improve the survival of pancreatic cancer.

Objective To compare MR,CT,and radiography in the detection of arthropathies in patients with hemophilia.Methods Forty-one symptomic joint images in the 14 patients with hemophilia, aged from 11 to 24 years,were used in this study.Each joint had the examinations of radiography,CT and MR within one day.The severity of each joint was staged using conventional radiographic classification. Severe HA patients with stage 5 were excluded from the study.Imaging findings of soft tissue swelling, osteoporosis,epiphyseal overgrowth,joint erosion,cyst,joint space narrowing,bone marrow,joint effusion, hemorrhage,synovial hypertrophy,widened intercondylar notch as well as anterior and posterior crueiate ligaments(only for knee joint)were used for the all imaging comparison.Results The 41 symptomatic joints in 14 patients with hemophilia were classified by radiographic criteria into stage 0(n=5),stage 1(n=7),stage 2(n=6),stage 3(n=8)and stage 4(n=15).Soft tissue swelling or joint effusion was observed in 33 joints by radiographs,in 34 joints by both CT and MR.Joint erosions were demonstrated in 34 joints by MR,in 33 joints by CT and 20 joints by radiographs.Joint cysts were shown in 21 joints by MR,in 18 joints by CT and 9 joints by radiographs.Significant differences in detection of erosion and cyst were found between radiography with either CT(P0.05).MR showed improvement for detecting nlore loci of both erosion and cyst than CT and radiography,and also CT showed the improvement than radiography.Bone marrow edema 14 joints, hemon'hage in 34 joints and synovial hypertrophy in 27 joints were revealed on MR images.Conclusion MRI is superior to CT and conventional radiography in detecting the abnormal changes and should be considered as the first choice among the imaging modafities in evaluating hemophilic arthropathies.

Objective To determine the distribution of HCV genotypes in patients with chronic hepatitis C,study the distribution of genotype in different gender and the relationship between genotypes and serum HCV-RNA levels.Methods Two hundred and six cases of HCV RNA positive patients(all with relevant clinical data) receiving pegylated interferon therapy were collected from May to December 2010.HCV RNA was detected in 206 hepatitis C patients from 40 hospitals in China by Roche Cobas AmpliPrep/Cobas TaqMan HBV test,and genotype was determined by Abbott RealTime HCV G enotype Ⅱ .The distribution of genotypes in the gender was analyzed by chi-square test analysis.The relationship between genotypes and serum HCV RNA levels was detected by single factor analysis and two independent sample t test analysis.Results There were seven different subtypes of HCV in 206 samples,including genotype 1,7 cases(3.4% ,7/206); genotype 1a,2 cases(1.0%,2/206); genotype 1b,123 cases (59.7 %,123/206); genotype 2,32 cases(15.5 %,32/206); genotype 3,27 cases(13.1%,27/206); genotype 6,6 cases(2.9% ,6/206) ;genotype 1/6,5 cases(2.4% ,5/206) ;genotype 2/4,1 cases(0.5%,1/206).There was no significant difference between HCV genotype and gender in 132 cases with genotype 1 and 65 cases with non-genotype 1(genotype 2,3,6) (x2 = 0.000,P ＞ 0.05).There was significant association between quantity of HCV RNA and genotype in 188 patients with HCV(F = 3.371,P＜ 0.01).The 197 patients with HCV single genotype were divided into five groups in terms of region(East,South,West,North and Center).There was no significant difference between HCV genotype 1 and non-genotype 1 in the five groups(x2 = 5.840,P ＞ 0.05).Conclusions It is suggested that HCV 1 b is the most prevalent type in China,followed by HCV 2.There is no significant difference between HCV genotype and gender.The levels of HCV RNA with genotype 1b are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 2 are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 6 are significantly higher than those with genotype 3.

OBJECTIVETo study the effects of two specific cyclooxygenase inhibitors (SCI), rofecoxib and celecoxib, combined with chemotherapeutic drugs 5-Fu, DDP and VP-16 on gastric cancer cell line BGC-823, and to evaluate whether specific cyclooxygenase inhibitors can be used as a synergetic agent in chemotherapy.

METHODSThe gastric cancer cell line BGC-823 cells were incubated for 48 hours with rofecoxib and celecoxib, 5-Fu, DDP and VP-16 (concentration gradient of 5-Fu, DDP and VP-16:1 microg/ml, 10 microg/ml and 100 microg/ml), or in combination, respectively. MTT working solution was added to each culture and calculated the survival rates of gastric cancer cells. Median-effect principle and Professor Jin's evaluation methods were applied to detect the interaction between the specific cyclooxygenase inhibitors and chemotherapeutic agents.

RESULTSThe inhibition rates of gastric cancer cells were 42.63% +/- 1.26% and 50.67% +/- 2.35% by treatment with 0.1 micromol/L rofecoxib and 50 micromol/L celecoxib, respectively. The inhibition rates of gastric cancer cells by treatment with 5-Fu, DDP and VP-16 at different concentrations (1 microg/ml, 10 microg/ml and 100 microg/ml) were 39.75% +/- 3.14%, 49.96% +/- 2.08%, 87.93% +/- 3.66%; 48.28% +/- 2.08%, 59.46% +/- 1.69%, 88.23% +/- 4.81%; and 29.23% +/- 3.27%, 49.34% +/- 3.75%, 79.24% +/- 2.44%, respectively. However, the inhibition rates showed a synergetic role while combined the two SCI (0.1 micromol/L rofecoxib and 50 micromol/L celecoxib) with chemotherapeutic agent at different concentrations (P <0.05).

CONCLUSIONBoth rofecoxib and celecoxib have an ability to suppress gastric cancer cells in vitro, and the synergetic role becomes evident when rofecoxib and celecoxib are combined with chemotherapeutic agents at different concentrations, which indicate that the two specific cyclooxygenase inhibitors may be used as a chemotherapeutic sensitizer.

Adenocarcinoma ; pathology ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Celecoxib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Cyclooxygenase Inhibitors ; pharmacology ; Dose-Response Relationship, Drug ; Drug Synergism ; Etoposide ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Lactones ; pharmacology ; Pyrazoles ; pharmacology ; Stomach Neoplasms ; pathology ; Sulfonamides ; pharmacology ; Sulfones ; pharmacology