s Objective To study the effect of HBV DNA of pregnant mothers on HBV intrauterine transmission. Methods Thirty one pairs of mothers and their newborn babies were detected for HBV by the HBV DNA qualitative PCR and HBV serology, as well as the concentration of HBV DNA in the blood by fluorescence quantitative PCR. Results There was a positive linear regression( r=0.5401, P=0.0017 ) between the concentration of HBV DNA in the infant and that in the mothers. Among the 31 infants, 19 showed abnormal by HBV DNA qualitative PCR and(or) HBV serology. It indicated that there was vertical transmission from the mothers to their infants. The transmission rate is 61%. The concentration of HBV DNA copies/ml(7.2?3.1) in mothers whose infants were abnormal was significantly higher than the others(5.8?1.3). Among the 31 mothers, 6 cases showed anti-HB positive and their babies were of much lower concentration of HBV DNA(4.2?1.8) than others(6.1?1.8). Conclusion The quantity of HBV in mothers reflected the strength of HBV vertical transmission and the chance of HBV transmission from mothers to infants have been decreased after mothers have obtained HbsAb.

Objective To use the enzyme linked immunosorbent (ELISA)to detect the hepatitis B virus (HBV)markers,and to perform the HBsAg quantitation and the HBV load detection for understanding the HBV carrying and viral replication situation when single HBcAb positive or both HBcAb and HBeAb positive.Methods The HBV markers HBsAg,HBsAb,HBeAg,HBeAb and HBcAb were detected with ELISA.1 098 cases of HBcAb positive,966 cases of both HBeAb and HBcAb positive and 832 cases of all HBV markers negative as control were selected and quantitatively re-detected HBsAg by using the chemiluminescence meth-od.The HBV load was detected by using the PCR method.Results Among 1 098 cases of single HBcAb positive,436 cases (39.7%)of HBsAg quantitation and 230 cases (20.9%)of PCR-DNA were detected out respectively;among 966 cases of both HBeAb and HBcAb positive,387 cases(40.1 %)of HBsAg quantitation and 212 cases(21.9%)of PCR-DNA were detected out re-spectively;among 832 cases of all HBV markers negative,6 case (0.7%)of HBsAg quantitation and 4 case (0.5%)of PCR-DNA were detected out respectively,there were statistically significantly differences among them (P < 0.05 ).Conclusion Adopting ELISA for detecting HBV markers,when single HBeAb positive or both HBcAb and HBeAb positive,HBsAg and the viral replica-tion are still be detected out,which needs to conduct further detection in order to avoid the medical risk due to the missed detection.

Objective To investigate the effect of HIF-2a silencing by transfection of siRNA into MG-63 cells un-der hypoxia. Methods HIF-2αexpression level in MG-63 cells under hypoxia was determined by Western Blot. Small in-terfering RNA (siRNA) was used to construct MG-63/siHIF-2α(siHIF-2α)cells and control MG-63/scramble (NC) cells. The expression levels of HIF-2α, Vascular endothelial growth factor (VEGF), p-Erk/ErK and Mcl-1 in MG-63, NC and si-HIF-2αcells was determined by Western Blot. NC and siHIF-2αcells were cultured under hypoxia. Cell viability was as-sessed by MTT assay. Migration was identified by scratch migration assay. Tumor formation was identified by clone formation assay. Nude mouse subcutaneous xenograft model was used to investigate tumor development in vivo. Results Hypoxia im-proved HIF-2αexpression in MG-63 cells in a time-dependent manner (F=2 037.412,P<0.001). HIF-2αexpression un-der hypoxia in siHIF-2αcells was lower than that in NC cells (P<0.01). Cell viability of siHIF-2αcells under hypoxia for 12 h and 24 h were lower than that in NC cells (P<0.05 or P<0.01). The relative width of scratch in siHIF-2αgroup under hypoxia for 12 h and 24 h were larger than that in NC group (P<0.01 or P<0.01). When cell counts reach 1 000-5 000, the clone formation rates of siHIF-2αcells were lower than that in NC cells (P<0.05 or P<0.01). The expression of VEGF, p-Erk/Erk and Mcl-1 protein under hypoxia in siHIF-2αcells was lower than that in NC cells(P<0.01). Tumor sizes, weights and density of siHIF-2α group in nude mice were suppressed compared with those in NC group (P<0.01). Conclusion Blocking HIF-2αsignal pathway warrants its investigation as a potential strategy in osteosarcoma treatment.

Objective To evaluate the efficacy and safety of Chushizhiyang ointment for the treatment of mild atopic dermatitis in infants.Methods A multicenter,randomized,open,active-controlled clinical trial was conducted.A total of 204 infants with atopic dermatitis were enrolled and randomly divided into 2 groups to be topically treated with Chushizhiyang ointment (test group,n =103) and hydrocortisone butyrate cream (control group,n =101),respectively,for 2 weeks.The improvement of eczema area and severity index (EASI) scores and quality of life was evaluated at 7 days and 14 days after the treatment,so was the incidence of adverse events and adverse reactions.Results Ninety-eight infants in the test group and 101 in the control group were included in the full analysis set,which revealed that the disease severity significantly decreased after the treatment in both groups.The EASI scores at the baseline and on days 7 and 14 were 2.47 ± 4.04,0.92 ± 1.25 and 0.39 ± 1.04 respectively in the test group,as well as 2.13 ± 2.01,0.85 ± 1.58 and 0.45 ± 1.65 respectively in the control group.Furthermore,the test group and control group both showed that EASI scores on days 7 and 14 significantly decreased compared with those at the baseline (the test group:T =-1 666,-1 793,respectively,both P ＜ 0.001;the control group:T =-1 951,-1 941,respectively,both P ＜ 0.001).No significant differences in EASI scores at the baseline or on days 7 and 14 were observed between the test group and control group (all P ＞ 0.05).The response rates in the test group and control group were 47.96％ (47/98) and 55.44％ (56/101) respectively on day 7,as well as 79.59％ (78/98) and 84.16％ (85/101) respectively on day 14,and there were also no significant differences between the two groups (both P ＞ 0.05).The adverse reactions mainly manifested as erythema,itching and scaling in the test group,as well as hypopigmentation,telangiectasia,scaling and hyperpigmentation in the control group.No significant difference in the incidence of adverse events was found between the test group (2.9％,3/103) and control group (6.9％,7/101).Conclusion Chushizhiyang ointment shows definite efficacy for mild atopic dermatitis in infants with good safety and tolerability,and can be a teatment option for mild atopic dermatitis in infants.

Objective To investigate the influence of health-care Qigong Baduanjin on mental health of patients with impaired glucose tolerance (IGT) . Methods Eighty-nine IGT patients were sampled from the physical examination center of Guangdong Provincial Hospital of Traditional Chinese Medicine, and then the subjects were randomly divided into groups A (N=30), B (N=29), C (N=30), which received health education, health education combined with walking exercise, health education combined with Baduanjin separately for one year. The scores of factors of physical and mental symptom checklist (SCL-90) were recorded before and after intervention. Results Compared to those before intervention, factor scores of interpersonal sensitivity and anxiety in group A were decreased obvi ously ( P<0.01) , factor scores of somatization, interpersonal sensitivity, depression, anxiety, terror, paranoia, psychosis in group B were decreased obviously (P<0.05 or P<0.01), and factor scores of somatization, inter-personal sensitivity, depression, anxiety, paranoia, psychosis in group C were also decreased obviously (P<0.05 or P<0.01) after intervention. Group B had lower factor scores of somatization and interpersonal sensitivity than group A (P<0.05), group C had lower factor scores of somatization, anxiety, and paranoia than group A (P<0.05 or P<0.01), group B had lower factor scores of anxiety than group C (P<0.05) . Conclusion Both Baduanjin and walking exercise can promote mental health of patients with IGT, and the effect of Baduanjin is superior to that of walking exercise.

Objective To investigate the features of brainstem auditoryresponses (BAR) in patients with attention deficit and hyperactive disorder (ADHD), mental retardation and conduct disorder. Methods BAR in children with ADHD ( n =37), mental retardation (MR, n =24) and conduct disorder (CD, n =22), and in normal children ( n =30) were measured by use of Nicolet Spirit electrophysiological instrument delivering stimulation of Clicks. Results The absolute latencies (AL) of wave Ⅲ、Ⅴ recorded from Fz region (WⅢ、Ⅴ/Fz) and wave Ⅲ from Pz region (WⅢ/Pz) as well as the absolute amplitudes (AA) of WⅢ/Fz and WⅤ/Pz were significantly different ( P

We amplified dhbC gene from the siderophore producing bacterium CAS15 by PCR. After ligated the PCR product to pMD18-T vector and then sequenced, we obtained a 1197 bp fragment. The blast result showed that the nucleotide acids of dhbC gene (Accession No. FJ194456) of CAS15 shared 99.7% identity with that of dhbC gene of Bacillus subtilis (GenBank Accession No. Z99120), and was predicted to encode a 43.8 kD polypeptide with 398 amino acid residues. We cloned the dhbC gene into expression vector pET-30a(+) and then transformed into Escherichia coli BL21(DE3) via calcium chloride transformation method, and obtained the recombinant E. coli BL21(DE3)/pET-30a-dhbC. Induced by 1 mmol/L IPTG the fusion protein 6His-DhbC, a 48.8 kD polypeptide was successfully expressed mainly in soluble form in E. coli BL21(DE3), and the amount reached highest at 30 degrees C for 4 h. According to the N-terminal fusion 6 His-tag, we purified the recombinant polypeptide by Ni2+ metal affinity chromatography and finally identified it by Western blotting. The result indicated that the recombinant DhbC had the antigenicity to rabbit anti-his-tag polyclonal antibody, which provides the basis for the study of practical utilization in production and the biocontrol mechanism of B. subtilis. Finally, we deleted dhbC gene by gene knockout and then retransformed it into the dhbC gene-delected mutant, which confirmed that dhbC gene play an important role in siderophore biosynthesis.
Bacillus subtilis ; enzymology ; genetics ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hydrolases ; genetics ; metabolism ; Hydroxybenzoates ; metabolism ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Siderophores ; metabolism

[ ABSTRACT] AIM: To investigate the influence of advanced glycosylation end products-modified bovine serum albumin (AGE-BSA) on mammalian target of rapamycin complex 1 (mTORC1), urokinase-type plasminogen activator re-ceptor ( uPAR) , and cell mobility in the podocytes, and to further explore the probable relationship.METHODS: The conditionally immortalized mouse podocyte cell line was cultured in vitro.MTT assay and immunofluorescence were used to analyze the cell viability and cytoskeleton of the podocytes treated with the stimuli and intervention agents.The activity of mTORC1 and the expression level of uPAR in normal podocytes and podocytes treated with control BSA or AGE-BSA were detected by Western blotting.The migration ability of the podocytes was determined by would-healing assay.Rapamycin was added to inhibit the activity of mTORC1 along with the addition of AGE-BSA to observe the changes of uPAR and the motility of podocytes.RESULTS:No significant difference of the cell viability or cytoskeleton in the podocytes treated with the stimuli and intervention agents was observed.AGE-BSA up-regulated the activity of mTORC1 and the expression of uPAR, and induced the high mobility of the podocytes.Rapamycin obviously reduced the high expression level of uPAR and the increase in the migration ability of podocytes caused by AGE-BSA treatment.CONCLUSION: AGE-BSA might cause the high migration of podocytes through the mTORC1/uPAR signaling pathway.

BACKGROUND:Hepatocyte transplantation has attracted more and more attention as a therapeutic measure for liver failure and genetic metabolic liver diseases.OBJECTIVE:TO evaluate the efficacy and safety of human hepatocyte transplantation in treating hepatitis B related liver failure in one case by a 2-year follow-up.DESIGN:A case-report of 2-year follow-up.SETTING:No.9 Department of Infectious Diseases,Bioengineering Research Room,the 302 Hospital of Chinese PLA.PARTICI PANT:One inpatient with hepatitis B related liver failure was selected from the 302 Hospital of Chinese PLA.and she was diagnosed according the laboratory tests.The transplanted hepatocytes were originated frOm the healthy liver of a 24-year-old man,who had signed the protocol for liver donation before death.METHODS:The hepatocyte transplantation was completed in the Department of Radiology,the 302 Hospital of Chinese PLA in December 2004.Liver was isolated to obtain human primary hepatocytes, and then cryopreserved.The hepatocytes were transplanted into recipient spleen via femoral vein after resuscitation.The clinical symptoms,changes of blood biochemical indexes,and changes of spleen MRI signals were observed before and after operation.The patient was reexamined every half a year after operation, including liver function, blood coagulation function,B-mode ultrasonography,gastroscopy and MRI,and she was followed up for 2 years. MAIN OUTCOME MEASURES:Liver function,blood coagulation function, imaging indexes, immunological indexes,complication and rejection.RESULTS:①Totally(1-2)×1010 hepatocytes were harvested,and the viability of rewarmed hepatocytes was 60%,and finally 2×109 hepatocytes were transplanted.②Two months later,the clinical symptoms of the recipient were obviously ameliorated,and serum bilirubin and aspartate aminotransferase(AST)were obviously decreased,while prothrombin activity was markedly increased.20 months later,the MRI results showed that there was hepatocyte image in spleen.Two years after operation.the total bilirubin level was 20 μmol/L,direct bilirubin level was 7 μmol/L, alanine aminotransferase was 416.75 nkat/L,AST was 533.44 nkat/L,albumin was 37 g/L,prothrombin activity was 90%,which were all obviously ameliorated as compared with those before operation(474.5 μmol/L,340.3 μmol/L,400.08 nkat/L,1 200.24 nkat/L,38 g/L,25%).The patient left the hospital 2 months later and could do light-burdened job.No complications of hydroperitonia and liver function failure, etc.were observed,and no rejection occurred.Several reexaminations by B-mode ultrasonography all indicated the further aggravations of liver cirrhosis and esophageal varices.She was admitted to hospital for twice because of esophageal varices bleeding,and cured by endoscopic variceal sclerosis therapy.CONCLUSION:Hepatocyte transplantation can ameliorate liver function without rejection,but it cannot relieve portal hypertension.

Objective To evaluate the efficacy and safety of desonide 0.05％ cream in the treatment of mild to moderate infantile atopic dermatitis (AD).Methods A multicenter,randomized,openlabeled,active-controlled clinical trial was conducted.A total of 120 infants with AD were enrolled from Department of Dermatology,Beijing Children's Hospital,Capital Medical University,Children's Hospital of Chongqing Medical University and Children's Hospital of Fudan University between December 2016 and November 2017.These patients were randomly divided into 2 groups to be topically treated with desonide 0.05％ cream (test group,n =61) and hydrocortisone butyrate 0.1％ cream (control group,n =59),respectively.After the treatment for 1,2 and 3 weeks,the response rate,improvement of eczema area and severity index (EASI) and infants' dermatitis quality of life (IDQOL) were evaluated,and adverse events and reactions were recorded.Statistical analysis was carried out with SAS 9.4 software using confidence interval method and non-inferiority test for the comparison of response rates between the test group and control group,two-sample t test for the comparison of quantitative data between two groups,paired t test for comparing pre-and post-treatment EASI scores and IDQOL scores.If the data were not normally distributed or there was heterogeneity of variance,Wilcoxon rank sum test was used for the comparisons of pre-and posttreatment EASI scores and IDQOL scores before and after the treatment,and chi-square test was used for the comparison of unordered categorical data between the test group and control group.Results At weeks 1,2and 3 after the initial treatment,analysis of the full analysis set (FAS) showed that 46 (86.79％),49(92.45％) and 51 (96.23％) patients in the test group received improvement respectively,and 36 (83.72％),35 (81.40％) and 41 (95.35％) in the control group received improvement respectively.The non-inferiority test showed that the response rate in the test group was non-inferior to that in the control group (non-inferiority boundary value was-0.15).At the baseline,1,2 and 3 weeks,the EASI scores in the test group were 4.57 ± 3.19,0.72 ± 0.89,0.45 ± 0.87 and 0.18 ± 0.40 respectively,and the EASI scores in control group were 4.50 ± 3.29,1.03 ± 1.81,0.62 ± 0.85 and 0.28 ± 0.82 respectively.Paired Wilcoxon rank sum test showed that the EASI scores in the test group and control group significantly decreased after the treatment for 1,2 and 3 weeks compared with those at the baseline (the test group:T =-715.5,-9.4,-715.5,respectively,all P ＜ 0.001;the control group:T =-437.5,-473.0,-472.0,respectively,all P ＜0.001).However,Wilcoxon rank sum test revealed that there was no significant difference in the EASI scores between the two groups at any of the above time points (P ＞ 0.05).Paired Wilcoxon rank sum test showed that IDQOL scores in the test group significantly decreased at the above time points after the treatment compared with those at the baseline (all P ＜ 0.001),while Wilcoxon rank sum test revealed that there was no significant difference in the IDQOL score between the test group and control group (all P ＞0.05).Adverse reactions were observed in 12 (19.7％) patients in the test group,and 10 (16.9％) in the control group,and there was no significant difference in the adverse reactions between the two groups (x2 =0.029,P ＞ 0.05).Conclusion The efficacy of desonide 0.05％ cream in the treatment of mild to moderate infantile AD is equivalent to that of hydrocortisone butyrate 0.1％ cream,so desonide 0.05％ cream can be a treatment option for mild to moderate infantile AD.