Objective:To evaluate the effect of anatomic resection (AR) and nonanatomic resection (NAR) on the prognosis of hepatocellular carcinoma (HCC) patients of China Liver Cancer Staging (CNLC) Ⅰa.Methods:PubMed, Embase and Cochrane Library databases were searched for articles on AR and NAR of CNLC Ⅰa stage HCC from January 2000 to March 2021. A meta-analysis was performed on patient characteristics, tumor characteristics, surgical procedures, postoperative pathological characteristics and long-term prognosis.Results:Of 9 case-control studies were included in this study, there were 2761 patients, with 1727 cases in the AR group and 1034 cases in the NAR group. When compared with the NAR group, the AR group had lower proportion of liver cirrhosis, lower ICG R15 value, higher proportion of Child-Pugh A grade, larger tumor diameter, higher proportion of microvascular invasion, longer operation time and higher intraoperative blood loss. The AR group had a better postoperative 5-year disease-free survival rate [ OR=1.54, 95% CI: 1.30-1.82, P<0.001] and a higher postoperative 5-year overall survival rate [ OR=1.27, 95% CI: 1.04-1.55, P=0.018]. Conclusion:AR is significantly better than NAR for preventing long-term recurrence and improving long-term survival in patients with CNLC Ⅰa stage HCC.

Flagellin can be expressed in monomeric or polymeric form based on assembly. The difference of these two forms of flagellin is less studied. In this experiment, recombinant plasmid pET-fliC/M2e2 was transferred into Escherichia coli BL21(DE3) and Salmonella SL5928 to express chimeric flagellin, mfliC/M and pfliC/M, respectively, and then their assembly characteristics were analyzed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis results indicated that the two recombinant bacteria could successfully express chimeric flagellin. The transmission electronic microscope observation showed that no flagella were found on the surface of recombinant E. coli, whereas it was found for recombinant Salmonella. After purification, distinct circular dichroism spectra between them were found and pfliC/M showed the similar structure as wild-type flagellin, but not for mfliC/M. The dynamic light scattering assay also indicated that the polymerization of mfliC/M was much lower than that for pfliC/M. Three hours after transfection into mouse peritoneal macrophages, both could induce interleukin 1β secretion, but mfliC/M is stronger than pfliC/M. These data will be helpful for the selection of expression form of flagellin.

Salmonella is an important zoonosis.China is the largest pork consumer.Contaminated pork is the main source of Salmonella disease at home and abroad.There are many swine Salmonella vaccine in use in Europe.Now the current swine Salmonella vaccine is a live attenuated vaccine,Salmonella can reduce colonization in pigs,and can induce immune response efficiently after the two immunizations.The control effect is poor,at this stage of swine vaccine cross protection research,type of Salmonella and vaccine antigen are not the same.This paper reviews the research progress of swine Salmonella vaccine.

Chronic neck pain is a common chronic musculoskeletal disorder. Research on neck muscle fatigue contributed to the understanding the pathogenesis of chronic neck pain. The influences caused by the neck muscle fatigue, such as the change of cervical proprioception and balance, could easily lead to chronic injury. Changes in morphology and recruitment strategy of cervical muscle could decrease muscle endurance, which bring about further damage. Targeted therapy and training can relieve muscle fatigue, improve muscle function, and reduce the recurrence rate. Surface electromyography has unique advantages in the evaluation of cervical muscle fatigue. The assessment of muscle fatigue could provide for assistance to the treatment and curative effect of chronic neck pain.

Objective To explore the mechanism of apoptosis after cryotherapy on tumors.Methods (1) GL261 glioma cells (1×107 cell/10 μL) were injected into the subcutaneous one ofC57 mice to establish tumor-bearing mouse models;when the diameter of tumor reached to 15-20 mm,the mice were randomly divided into cryogenic treatment group and sham-operated group (n=1 0);mice in the cryogenic treatment group were given surgical cryotherapy,while those in the sham-operated group only performed surgery without cryotherapy.TUNEL was used to detect the cell apoptosis in glioma tissues 12 and 24 h after operation;and Western blotting was employed to detect the protein expressions of pro-caspase-8,pro-caspase-9 and poly-ADP-ribose polymerase (PARP).(2) GL261 glioma cells were divided into control group and one time cryogenic release group,and DMEM and one time of cryogenic release were given to the two groups,respectively;12 h after the treatment,TUNEL was used to observe the cell apoptosis in glioma tissues,and Western blotting was employed to detect the protein expressions.Results (1) TUNEL indicated that the cells in the S1 region of the glioma tissues from mice in the cryogenic treatment group were uniformly died;significant apoptosis was noted in cells of the S2 region at 12 h after treatment;while,24 h after treatment,S1 region still showed uniform necrosis,S2 region showed apoptotic regression,and S3 region showed new apoptosis at the target side.Western blotting indicated that pro-caspase-9 and PARP protein expressions at the S2 region were signficantly decreased as compared with those at the S1,S3 and S4 regions (P＜0.05),and pro-caspase-8 protein expression at the S3 region were signficantly reduced as compared with those at the S1,S2 and S4 regions in the cryogenic treatment group (P＜0.05).(2) TUNEL showed that significantly increased GL261 glioma cell apoptosis rate was noted in the one time cryogenic release group as compared with that in the control group (P＜0.05);Western blotting indicated that as compared with the control group,the cryogenic release group had significantly decreased pro-caspase-8 and pro-caspase-9 expressions (P＜0.05).Conclusion Cryogenic release or substances released from tumor tissues after cryoablation shows an effect on promoting apoptosis.

Two hundreds postmenopausal women with osteopenia, aged 45-80, were randomly divided into 4 groups, and received different drug interventions. After treatment for 12 months, the lumbar spine bone mineral density(BMD)and total hip BMD in alendronate group increased significantly(3.5% and 2.6%, both P＜0.05), the lumbar spine BMD raised 2.0% in selective estrogen receptor regulator group(P＜0.05). Bone turnover indices improved in both groups(all P＜0.05). No change in BMD or bone turnover indices was found in patients treated with active vitamin D3and calcium.

Bovine interferon-gamma (BoIFN-gamma) gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from total RNA of bovine spleen lymphocytes stimulated with ConA. The products of RT-PCR were cloned into pVAX1 vector, positive recombinant clone was identified by restriction enzyme digestion and sequencing. The recombinant plasmid pVAX1-BolFN-gamma was transfected into COS-7 cells mediated by lipofectine, indirect immunofluorescent assay analysis confirmed that rBoIFN-gamma was expressed in COS-7 cells. BoIFN-gamma gene (without signal peptide) was cloned into pET-30a(+) and pGEX-6p-1 vector, and transformed into the Escherichia coli cells. After optimizing the induction condition, SDS-PAGE analysis showed that the expression products were all found in soluble form and had a molecular weight of 23 kDa and 43 kDa respectively. BoLFN-gamma precursor gene (with signal peptide) was cloned into transfer vector pFastBac 1, and transformated into DH10Bac E. coli cells. By site-specific transposition, BoIFN-gamma gene was integrated into shuttle vector Bacmid, and transfected into the Sf9 insect cells mediated by lipofectine to produce recombinant baculovirus. Indirect immunofluorescent assay analysis confirmed that rBac-BoLFN-gamma was expressed successfully in Baculovirus vector system. The antiviral activities of rHis-BoIFN-gamma, rGST-BoIFN-gamma and rBac-BoIFN-gamma were up to 8.389 x10(7) U/mg, 6.554 x10(5) U/mg and 4.096 x 10(4) U/mL respectively, which were analyzed in MDBK/VSV system. A sandwich ELISA was established using monoclonal antibodies 3E6 and 5G4, which can detect BoIFN-gamma in quantity and provide a useful method for the clinical practice and research of BolFN-gamma.
Animals ; Antiviral Agents ; pharmacology ; Baculoviridae ; genetics ; metabolism ; COS Cells ; Cattle ; Cercopithecus aethiops ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Interferon-gamma ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Transfection

The polymorphisms of paraoxonase 1 ( PON 1 ) and ApoE gene in 192 patients with type 2diabetes mellitus (DM group) , 116 cases of atherosclerosis (AS) without DM (AS group), and 105 normal subjects (NC group ) were detected by means of PCR-restriction fragment length polymorphism. The serum concentration of Ox-LDL was determined by enzyme linked immunosorbent assay. The results showed that plasma Ox-LDL level in DM group was higher than those in AS and NC groups[(754.2±279.9 vs 526. 1±186.2 and 421.1 ± 163.2 )μg/L, P<0.01]. Ox-LDL level in the patients with QQ genotype of PON 1 was higher than those in QR and RR genotypes in type 2 DM[(846.6±147.5 vs 763.4±126. 7 and 7 1 3.2 ±132.4 ) μg/L, P<0. 0l],and Ox-LDL level in the patients with ε3/4+ ε4/4 genotype of ApoE was also higher than those in ε3/3 and ε2/2 + ε2/3 geneotypes in type 2 DM[( 824.3 ± 173.5 vs 741.6± 182. 5 and 718.3 ± 167.5 ) μg/L, P<0.05], but the difference was not found in atherosclerosis without DM. Multiple regression analysis showed that disease duration, PON1 and ApoE genotypes were the independent predictors of plasma Ox-LDL level in patients with type 2 DM.

Objective To investigate the risk factors for postmenopausal osteoporosis in Shanghai.Methods Five hundred postmenopausal community-dwelling women aged 45-80 years were recruited. The case-control study was performed from June 2008 to September 2008.A total number of 250 women with postmenopausal osteoporosis identified with their bone mineral density (BMD) were selected as case group, and 250 non-osteoporosis women were selected as control group. BMD was measured by dual energy X-ray absorption (DEXA). Results Univariate logistic regression analysis showed that age, eduction level, occupation, years since menopause, BMI, use of calcium,historyofnon-violencefracture,fall,diabetesmellitus,chronicgastricdiseases, gastrointestinal resection and diarrhea were related to osteoporosis.Multiple logistic regression showed that age, years since menopause and nutritional status were the risk factors for osteoporosis. ConclusionsThe occurrence of osteoporosis is related with many factors in postmenopausal women in Shanghai, and women with early menopause, low BMI and older age should pay more attention to the prevention and treatment of osteoporosis.

Objective To determine the immune responses induced by recombinant Salmonella ty-phimurium expressing the secreting antigen ESAT-6 of Mycobacterium tuberculosis. Methods ESAT-6 cod-ing gene was cloned and identified by PCR and sequencing. Prokaryotic expression plasmid pYA33-esat car-rying the ESAT-6 coding sequence was constructed firstly and electro-transformed into an attenuated strain X4550 of Salmonella typhimurium, the recombinant bacteria was named as X4550(33-esat). C57BL/6 mice were immunized intranasally (I. N) with 108 CFU recombinant bacteria at day 0 and 18. Cells from spleen, lung, mesenteric lymph node (MLN) and Peyer's patch (PP) were collected from mice after second immu-nization, and the specific IFN-γ-secreting cells and IL-4-secreting cells were detected by ELISPOT assay u-sing ESAT-6 peptide as stimulus. Furthermore, CTL effects were in vivo evaluated by CFSE assay. Results The results showed that cellular immune responses specific for ESAT-6 could be detected by ELISPOT assay. In lung and PP cells, immune responses against ESAT-6 were biased toward Th1 type, the frequency of IFN-γ-secreting cells was much higher than that of IL-4-secreting cells. In splenocytes and MLN cells, the anti-gen specific immune responses acted as Thl and Th2 balance, the frequency of IFN-γ-secreting cells was close to that of IL-4-secreting cells. CFSE assay indicated that recombinant bacteria could induce the high level of CTL effects specific for ESAT-6 peptide. Conclusion These results suggested that recombinant Sal-monella typhimurium X4550(33-esat) not only can induce cellular immune responses, but also can elicit specific CTL responses after I. N immunization. It also provided the useful information for the control of infec-tious disease of tuberculosis.