SHIV-CN97001 played an important role in assessing the immune effect and strategy of the AIDS vaccine which included genes of the predominant prevalent HIV-1 strain in China. In this study, SHIV-CN97001 was in vivo passaged serially to construct pathogenic SHIV-CN97001/rhesus macaques model. To identify variation in the gp120 region of SHIV-CN97001 during passage, the fragments of gp120 gene were amplified by RT-PCR from the plasma of SHIV-CN97001 infected animals at the peak viral load time point and the gene distances (divergence, diversity) were calculated using DISTANCE. The analysis revealed that the genetic distances of SHIV-CN97001 in the third passage animals were the highest during in vivo passage. It had a relationship between viral divergence from the founder strain and viral replication ability. The nucleic acid sequence of the V3 region was highly conservative. All of the SHIV-CN97001 strains had V3 loop central motif (GPGQ) and were predicted to be using CCR5 co-receptor on the basis of the critical amino acids within V3 loop. These results show that there was no significant increase in the genetic distance during serial passage, and SHIV-CN97001 gp120 gene evolved toward ancestral states upon transmission to a new host. This could partly explain why there was no pathogenic viral strain obtained during in vivo passage.

Objective To study the expression and prognostic significance of neuroendocrine differentiation markers chromogranin A(CgA)and synaptophysin(SYN) in patients with resectable non-small cell lung cancer(NSCLC).Methods From January 2000 to January 2003,123 patients with NSCLC who received operations were investigated.The resected specimens and clinical data were collected.Immunohistochemical Elivison method was used to detecte the expression of CgA and SYN.Kaplan-Meier survival curve and Cox proportional hazard model multivariate analysis were applied for the prognostic factors. Results The positive expression rates of CgA and SYN were 22%,17.9%,respectively.The expression of SYN was associated with histological differentiation(P=0.001).No significant association was found between NSCLC with neuroendocrine differentiation(NSCLC-ND) and sex,age,smoke index,TNM Stage and pathology classification.No evidence showed the patients with positive expression of CgA or SYN could be tolerant with more cycles of chemotherapy(P=0.406).Kaplan-Meier survival curve indicated the survival had a relation with the expression of CgA and SYN.It was revealed by Cox analysis that SYN(P=0.001),TNM stage(P=0.02)and the maximal diameter of tumor(P=0.049) were independent prognostic factors. Conclusion The patients with NSCLC-ND had a poorer prognosis.SYN may be one of the prognostic factors in patients with NSCLC.

Objective To detect the expression level of aquaporin 8 (AQP8) in patients with functional constipation(FC) or constipated irritable bowel syndrome (IBS-C),and the correlation between the expression of AQP8 and clinical features.Methods From March to December 2014,a total of 16 patients with IBS-C and 19 patients with FC met Rome Ⅲ criteria were collected,and nine healthy individuals were assigned to control group at the same period.The ascending and decending colonic tissues mucosa of FC,IBS-C and control group were taken under endoscope.The expression of AQP8 at mRNA and protein level was detected by real-time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).The differences in AQP8 mRNA expression and AQP8 relative area were analyzed by Kruskal-Wallis test among groups,and Pearson correlation coefficient was performed for correlation analysis between the expression and clinical features.Results The relative expressions of AQP8 mRNA of ascending colon and descending colon of FC patients (1.38,0.61 to 4.09;2.65,0.82 to 7.52) and IBS-C patients (2.23,0.82 to 4.67;1.35,0.51 to 2.03) were higher than those of control group (0.56,0.19 to 0.97;0.38,0.21 to 1.19),and the differences were statistically significant (ZFc =-2.435,-3.149,ZIBS-C =-2.690,-2.152;all P＜0.05).AQP8 mRNA expression of descending colon in patients with FC was higher than that of patients with IBS-C,and the difference was statistically significant (Z =-2.003,P =0.045).The expression of AQP8 in patients with FC and IBS-C was positively correlated with disease course (ascending colon r=0.57 and 0.53;descending colon r=0.49 and 0.54,all P＜0.05),and was negatively correlated with frequency of defecation (ascending colon r=-0.82 and-0.61;descending colon r=-0.49 and-0.53,all P＜0.05).There was no correlation between the expression of AQP8 and age,gender,onset age,presence of abdominal symptoms of the patients (all P＞ 0.05).Most of AQP8 of FC group was expressed in cytoplasm of colonic mucosa epithelial cells,while that of IBS-C group and control group was mostly expressed at apical membrane and basal membrane of epithelial cells.The results of semi-quantification demonstrated that AQP8 relative area of descending colon of FC and IBS-C group increased compared with that of control group (3.42％ (1.24％ to 5.61％),2.45％(1.72％ to 4.27％) vs 1.18％ (0.35％ to 2.81％);Z=-2.534,-2.151,both P＜0.05).Meanwhile,AQP8 relative area of ascending colon of FC group increased compared with that of control group (2.46％(1.48％ to 4.18％) vs 1.14％(1.29％ to 2.15％) Z=-2.041,P＜0.05).Conclusion There are differences in AQP8 expression quantity and location in cells of descending colon between patients with FC and patients with IBS-C,which is a way for differentiation these two diseases.

Objective To study the effects of different level of iodine nutrition on the thyroid function in women of reproductive age. Methods A total of 100 (50 from each) women of reproductive age but not pregnant were collected from iodine deficient and adequate areas. The questionnaire was obtained individually with items concerning personal history of thyroid diseases, goiters and category of edible salt and drinking water based on the project design. The household salt and drinking water were collected for measuring iodine content, and blood samples were obtained for TSH, FT4 and FT3 testing. Results The coverage of iodized salt and the median level of urinary iodine in iodine deficient women(72.0% and 95.5 μg/L) were obviously lower than that in iodine sufficient women(100.0% and 167.4 μg/L, χ2=16.28, U = 632.00, P < 0.01). Median level of serum TSH in iodine deficient women (2.56 m U/L) appeared in an increasing tendency compared to the iodine sufficient women (1.88 mU/L), but there was no significance (U=990.50, P > 0.05). Serum FT4 mean level in iodine deficient women [(14.7±2.0) pmol/L]was lower than that in iodine sufficient women[(17.0±3.8)pmoI/L, t=3.76, P<0.01]. There was no difference in serum FT3 between two group women[(5.1±1.4), (4.8±0.5)pmoI/L, t = 1.59, P > 0.05]; but FT3/FT4 ratio in iodine deficient women(0.33±0.04) was markedly higher than that in the iodine sufficient women(0.30±0.04, t=3.13, P<0.01). The percentage of thyroid dysfunction in iodine deficient women[20.0% (10/50)]was higher compared with the iodine sufficient women[8.0%(4/50)], but without significance(χ2=2.99, P>0.05). Conclusions Iodine deficiency is a primary cause leading to hypothyroid in women of reproductive age. Long term of iodized salt usage is an efficient way to correct iodine deficiency.

Objective :To study effects of early rehabilitation after percutaneous coronary intervention (PCI) on car‐diac rehabilitation in patients with coronary heart disease (CHD).Methods :A total of 100 CHD patients undergoing PCI in our hospital were randomly and equally divided into routine nursing group and early rehabilitation group . Clinical indexes and incidence rate of major adverse cardiovascular events (MACE) after PCI ,scores of Morningside rehabilitative state scale (MRSS) ,self‐rating anxiety scale (SAS) ,self‐rating depression scale (SDS) ,social disabili‐ty screening schedule (SDSS) ,short‐form 36 health survey questionnaire (SF‐36) before and after treatment and pa‐tient satisfaction were compared between two groups before discharge .Results : Compared with routine nursing group ,there were significant reductions in first‐time get out of bed time 、 exercise time ,complete self‐care time , hospitalization time ,postoperative incidence rate of MACE (52% vs .18%) in early rehabilitation group , P＝0. 001 all ;After treatment ,compared with routine nursing group , there were significant reductions in scores of MRSS [ (5.87 ± 1.21) scores vs .(2.69 ± 1.01) scores] ,SAS [ (49.96 ± 3. 98) scores vs .(44.56 ± 3.12 ) scores] ,SDS [ (49.89 ± 3.85) scores vs .(45.38 ± 3.15) score] and each dimension score of SDSS ,and significant rise in SF‐36 score [ (89.76 ± 7.23) scores vs .(93.98 ± 8. 09) scores] and total satisfaction rate of patients (70% vs.90%) in early rehabilitation group , P＜0. 05 or ＜0.01. Conclusion :Early rehabilitation after PCI can significantly improve cardiac rehabilitation level ,psychological state ,social function and quality of life ,shorten postoperative recovery time and comprehensively improve patient satisfaction in CHD patients .

BACKGROUNDRevision anterior cruciate ligament (ACL) surgery can be expected to become more common as the number of primary reconstruction keeps increasing. This study aims to investigate the factors causing instability after primary ACL reconstruction, which may provide an essential scientific base to prevent surgical failure.

METHODSOne hundred and ten revision ACL surgeries were performed at our institute between November 2001 and July 2012. There were 74 men and 36 women, and the mean age at the time of revision was 27.6 years (range 16 - 56 years). The factors leading to instability after primary ACL reconstruction were retrospectively reviewed.

RESULTSFifty-one knees failed because of bone tunnel malposition, with too anterior femoral tunnels (20 knees), posterior wall blowout (1 knee), vertical femoral tunnels (7 knees), too posterior tibial tunnels (12 knees), and too anterior tibial tunnels (10 knees). There was another knee performed with open surgery, where the femoral tunnel was drilled through the medial condyle and the tibial tunnel was too anterior. Five knees were found with malposition of the fixation. One knee with allograft was suspected of rejection and a second surgery had been made to take out the graft. Three knees met recurrent instability after postoperative infection. The other factors included traumatic (48 knees) and unidentified (12 knees).

CONCLUSIONTechnical errors were the main factors leading to instability after primary ACL reconstructions, while attention should also be paid to the risk factors of re-injury and failure of graft incorporation.

Adolescent ; Adult ; Anterior Cruciate Ligament Reconstruction ; adverse effects ; Female ; Humans ; Joint Instability ; etiology ; Male ; Middle Aged ; Retrospective Studies

Objective To evaluate the efficacy and safety of acute ureteroscopy for the treatment of ureterie stones during middle and late pregnancy.Methods From June 1998 to March 2005,17 pregnant women(mean age,27 years;age range,21-35 years)with ureteric stones were treated by ureteroscopy when the fetus was at 20-36 weeks of gestation(mean,29 weeks).All the cases presented with urgent symptoms such as recurrent renal colic(11 cases),fever(4)or acute obstructive anuria(2).Among 17 cases,the stones(between 6 mm?7 mm and 13 mm?21 mm)were located in the upper(8 cases),middle(5)or lower ureter(4);and on the left side(5 eases),on the right(10)and on both(2)of the lower ureter. Mild hydronephrosis were observed in 6 cases and moderate hydronephrosis in 11,Of the 17 cases,14 under- went ureteroscopic pneumatic lithotripsy;in 1 case the calculi were pushed to the renal pelvis;and 2 cases were treated by Double-J catheter drainage.Results All the urgent symptoms in 17 cases were relieved after treatment.The stone-free rate of initial treatment was 82.4%(14 of 17).Three cases with residual stones were treated by Douhle-J catheters,which were replaced every 3 months until the calculi were re- moved.No abortion,premature delivery or complications such as ureter perforation occurred.Mild renal colic occurred in 1 case after insertion of Douhle-J catheter,and it was relieved 3d later;gross hematuria occurred in l case and disappeared 6 d later without treatment.All 17 patients had normal delivery and gave birth to healthy children.Conclusions Ureteroscopy is a safe and reliable method for the treatment of ureteric calculi during middle and late pregnancy.

OBJECTIVETo study the specific cytotoxicity mediated by anti-P-gp/anti-CD(3) diabodies in multidrug resistant solid tumor using P-gp as target.

METHODSThe anti-P-gp/anti-CD(3) diabodies were secreted from E. coli strain 16C9 containing the expression plasmid PAYZDCP, grown at 30 degrees C in a shaker flask; the diabodies were purified by affinity chromatography and identified by SDS-PAGE; the effect of the anti-P-gp/anti-CD(3) diabody mediated lysis of P-gp-expressing tumor cells was assayed by (51)Cr release assay in vitro, and by human KB nude mouse xenograft models in vivo.

RESULTSThe diabodies were generated by bacteria as a soluble functional form and purified by one-step affinity chromatography with a yield > 4 mg/L culture medium. In (51)Cr release assay, the diabodies targeted human activated T cells to lyse P-gp(+)-KB/MDR cells in a dose-dependent manner. It suggested that the diabody was able to induce an efficient lysis of the target cells by human T cells in vitro. When combined with activated human T cells, the diabody significantly inhibited the growth of KB/MDR, but had no effect on KB xenografts.

CONCLUSIONThe anti-P-gp/anti-CD(3) bispecific antibody is a potent agent for targeting human T lymphocytes to lyse solid tumor cells overexpressing P-gp in vitro and in vivo.

ATP-Binding Cassette, Sub-Family B, Member 1 ; immunology ; Animals ; Antibodies, Bispecific ; immunology ; therapeutic use ; CD3 Complex ; immunology ; Drug Resistance, Multiple ; immunology ; Drug Resistance, Neoplasm ; immunology ; Female ; Humans ; KB Cells ; Mice ; Mice, Nude ; Neoplasms, Experimental ; therapy ; Protein Engineering ; methods ; Recombinant Proteins ; therapeutic use ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo explore the effects of survivin antisense RNA on taxol-induced apoptosis in leukemia cell line HL-60.

METHODSA survivin antisense eukaryotic vector pcDNA3-SVVas was transferred into HL-60 cells by electroporation. The live fraction was determined by trypan blue dye exclusion assay. Cell counting and MTT assay were performed to evaluate the sensibility of the transfected cells to taxol. Apoptosis was detected by DNA gel electrophoresis and nuclear staining.

RESULTSTwo positive cell clones, HL-60 SVVas and HL-60 neo were obtained. Compared to HL-60 and HL-60 neo cells, HL-60 SVVas cells growth was significantly reduced (P < 0.05). By MTT assay, the IC(50) of taxol to HL-60 SVVas, HL-60 neo and HL-60 cells were (14.4 +/- 1.87) ng/ml, (31.9 +/- 6.38) ng/ml and (32.0 +/- 3.52) ng/ml, respectively, the difference was significant by statistic analysis (P < 0.01). Agarose gel electrophoresis of genomic DNA from HL-60 SVVas showed typical DNA ladder, but DNA from HL-60 neo and HL-60 did not. Nuclei become condense in HL-60 SVVas cells.

CONCLUSIONSurvivin antisense RNA could enhance taxol-induced apoptosis in leukemia cell line HL-60. This may lay an experimental foundation for further research of gene therapy in leukemia.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Drug Synergism ; HL-60 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Transfection

BACKGROUNDThere is still a paucity of data on hepatitis B virus (HBV) subgenotype prevalence in North China based on sequencing of large-size samples. In addition, whether HBV genotypes impact drug-resistance-associated and HBV e antigen (HBeAg)-loss-associated mutations in patients with chronic hepatitis B (CHB) is still under investigation. This study aimed to disclose clinical prevalence of HBV genotypes/subgenotypes in North China and the clinical implications of HBV genotype classification in respect to HBeAg loss and drug-resistant occurrence.

METHODSSera were collected from 1301 nucleos(t)ide analog-experienced CHB patients. Viral DNA was extracted and used as template for HBV genome amplification by nested PCR. DNA sequencing was performed for the analysis of HBV genotypes/subgenotypes, drug-resistance-associated mutations in polymerase gene and HBeAg-loss-associated mutations in precore/basal core promoter (BCP) regions.

RESULTSHBV/B, HBV/C, and HBV/D were detected in 190 (14.6%), 1096 (84.2%), and 15 (1.2%) patients, respectively. HBV/B2 (182/190), HBV/C2 (1069/1096), and HBV/D1 (12/15) were predominant subgenotypes within individual genotypes. By contrast, C2 prevalence is relatively lower in Beijing area (77.2%) than in other north areas (84.9% - 87.4%). HBV/C-infected patients had an older age and a lower serum albumin level but similar HBV DNA and alanine aminotransferase (ALT) levels compared to HBV/B-infected patients. HBV/C infection had a higher incidence of lamivudine-resistant mutations rtM204I/V (44.9% vs. 30.2%, P < 0.01) and BCP mutations A1762T+G1764A (65.8% vs. 40.0%, P < 0.01) compared with HBV/B infection.

CONCLUSIONSC2 is the most prevalent HBV subgenotype followed by B2 in CHB patients in North China; and HBV genotype prevalence is influenced by immigrant population. HBV/C infection is likely to have longer disease duration and severer liver functional impairment and might be more susceptible to develop lamivudine resistance compared to HBV/B infection.

Adult ; Antiviral Agents ; therapeutic use ; China ; DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Female ; Genotype ; Hepatitis B virus ; classification ; drug effects ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Middle Aged ; Mutation