Objective To study effect of excessive iodine intake on sodium-iodide symporter(NIS)mRNA and protein expression of breast in lactating rats.Methods60 Wistar rats,having been weaned for one month,were randomly divided into three groups according to their body weights,I.e,①normal iodine(NI,30 rats);②ten fold high iodine(10 HI,15 rats);③one hundred fold high iodine(100 HI,15 rats).Eating food containing iodine of 300μg/L and drinking water of iodine at 5,1845,20 295μg/L,respectively.After fed for 3 months,the rats mated and had offspring,and urine and milk iodine of lactating rats were determined by As-Ce-catalytic spectrophotometric method.Their marmnary glands were sampled at lactation day 10.Then NIS mRNA expression by RT-PCR was determined and NIS protein by immunohistochemistry(SABC)was observed.Results The urine iodine of 10 HI group(3597.5μg/L)and 100HI group(25 404.3μg/L)increased obviously compared with that of NI group(344.7μg/L).The milk iodine of 10HI group(27.1×103μg/L)and 100HI group(191.0×1μg/L)was higher than that of NI group(6.0×103μg/L),but the increased fold of milk iodine was not paralleled with that of urine iodine.Difference of NIS mRNA expression was significant(F=24.19,P＜0.01)among the groups,and the NIS mRNA expression in 10HI(1.250±0.034)and 100HI(1.272±0.039)group were less than that in NI (1.532±0.044)group(P＜0.01).The breast NIS mRNA expression in lactating rats(1.532±0.044)was significantly higher than that in unlactating rats(0.879±0.018,P＜0.01).With the increasing iodine uptake,NIS protein expression decreased.Conclusions The NIS mRNA and protein in rat breasts is down-regulated by excessive iodine intake.So increasing extent of milk iodine concentration is inhibited,which is important to prevent off-spring from getting excessive iodine intake from parental generation.

Objective:To analyze the prevalence of syphilis infection and influence factors of syphilis infection among the male STD attendants in Songyuan City of Jilin Province, and to provide theoretical basis for the development and implementation of syphilis intervention policy.Methods:A questionnaire survey and serological tests were conducted among the men who went to STD clinic for treatment in the ZhanDong Dermatology Specialist Hospital or Qianguo County Hospital in Songyuan City from 2011 to 2015.Chi-square test was performed to compare the prevalences of syphilis infection between different groups.Multivariable Logistic regression model was used to find the independent factors of syphilis infection among the male STD attendants.Results:Of all 2 000 male STD attendants who involved in the study,the mean age was (34.50±9.03)years old.218 persons (10.90%)of them had sexual behavior with female sex workers in the last three months,433 persons (21.65%)of them had temporary sexual behavior in the last three months,42 persons (2.10%)of them had homosexual behavior in the last three months.Of the respondents,238 persons (11.90%)had a previous diagnosis of sexually transmitted diseases,86 persons of them had gonorrhea (36.13%),43 persons had syphilis (18.07%),15 persons had genital tract Chlamydia (6.30%),55 persons had genital herpes (23.11%),40 persons had genital herpes (16.81%),and 1 person had combined infection of gonorrhea and syphilis. The prevalence of syphilis of 2 000 male STD attendants was 6.4%.From 2011 to 2015,there was a decrease in the prevalences of syphilis (χ2 =44.25,P <0.001).In the last 3 months, the risk of the male STD attendants who had commercial sex with prostitute,temporary sex behavior,homosexual behavior and a history of STD infection to be infected with syphilis in the past were 3.75 times (OR= 3.75,95% CI:2.46 - 5.71),2.31 times (OR = 2.31,95% CI: 1.56 - 3.41),2.97 times (OR=2.97,95%CI:1.33-6.64)and 1.69 times (OR=1.69,95%CI:1.07-2.67)than those who did not have those behaviors,respectively. Conclusion:There is a significantly decreasing tendency in the prevalence of syphilis among the male STD attendants in Songyuan City in the past 5 years.The high risk sexual behaviors are the main influence factors of syphilis infection among them.

Objective:To establish a method of multi-PCR to amplify the complete DNA sequence (CDS) of TCR ? and ? chain of the antigen-specific T lymphocytes in local pathologic specimen of active pulmonary tuberculosis patients, and to analyze ?/? T cell receptor gene rearrangement and CDR3 repertoire.Methods:The lymphocytes in bronchoalveolar lavage (BAL) of active pulmonary tuberculosis patients were separated. Following total RNA extraction, cDNA synthesis, Multi-PCR, recombinant clones construction, and sequencing, the CDS of TCR ? and ? chains from these lymphocytes were analyzed by using software of DNAstar and internet TCR resources.Results:24 of ? chain CDS and 13 of ? chain CDS from 3 samples of BAL were obtained. As for TCR ? chain, AV1S2 (54%), AV12S3 (41%), and AV12S2(5%) appeared frequently. BV2(38%), BV29S1(46%), BV14(3%), and BV4S2(3%) in TCR ? chain appeared more often. There were CDR3 diversities between samples and even in the same sample by amino acid sequence analysis, but there were a few identical or similar amino acid sequences. There was the same amino acid sequence of SVGTGTLHQETQY in CDR3 region of ? chain of BAL sample No.1 and No.2; The sequence of AVRDWAGNMLT appeared in two ? chains of BAL sample No.2 and No.3; Moreover, the sequence of AV…DNN…RLM appeared in ? chains of BAL sample No.2 and No.3.Conclusion:A method of Multi-PCR is used to amplify TCR ? and ? chain CDS of tuberculosis patients. There are characteristic T cell clones to proliferate,with TCR ? and ? chain repertiore skewing in local infective focus. The sequences of CDR3 in different TCR clones are mostly different but there are a few identical or similar sequences in the same patient or even between different patients. The identical amino acid sequences of CDR3 are possibly specific for recognizing MTB polypeptide.

Objective To investigate the inhibition effect of Ki67 AS-ODN on pr ostate carcinoma PC-3 cells,and its possible synergism existing in combination of Ki67 AS-ODN and paclitaxel.Methods Ki67 AS-ODN were transfected into PC-3 cells by lipofectamine. Cell proliferation was measured by CCK8 method,and Ki6 7 mRNA expression was detected by RT-PCR. The synergetic effect of Ki67 AS-ODN combined with paclitaxel was evaluated by Zhengjun Jin Q method. Results AS-OD N at the concentration of 31.25 nmol /L can significantly inhibit PC-3 cells pr oliferation(P

Objective To explore the outcomes of the transplanted kidney as donor for clinical renal transplantation and summarize experience in combination with related literature.Method This study retrospectively analyzed the clinical documents of one case of uremia receiving renal allograft transplantation with the transplanted kidney as the donor in one case of renal transplantation after brain death in February,2015.The donor was a 31-year-old man who received renal transplantation for uremia in November,2014 and obtained normal renal function.Two months later,the patient was brain dead because of neurologic disorder and donated his transplanted kidney.The serum creatinine of the donor was 167 μmol/L,and the glomerular filtration rate was about 35 mL/min befor donation.The recipient was 27 years old who needed transplantation because of chronic renal function failure and uremia.Preoperation tests showed that PRA was negative,and serum creatinine was 1 353 μmol/L.After separating and dissecting the donor kidney carefully,we perfused and compensated the kidney by Lifeport Organ Perfusion and Preservation Conveyor.The warm ischemia time was about 15 min.The renal vein of the donor was anastomized with right external iliac vein of the receptor,artery with right external iliac artery,and ureter with right centrifugal ureter.Result The operating time was more than 3 h.Postoperatively,the recipient was given the immunosuppressive regimen as tacrolimus,mycophenolate mofetil and methylprednisolone to prevent rejection.At 1 st day postoperation,the 24-h urine volume of the receptor was 5 000 mL,serum creatinine was declined gradually to a minimum of 180μmol/L,and there was trace urine protein.The renal function of patient recovered well by now.Meanwhile,the patient was still under the follow-up.Conclusion It is practical that using transplanted kidney as donor kidney for re-transplantation.There were certain clinical significance for shortening the waiting time of renal transplantation in uremia patients and relieving the shortage of transplant kidney.

Objective To construct and apply a cell line screening Mycobacterium tuberculosis (Mtb)-specific tetramers of CD4+α/β T cell receptor(TCR). Methods The β chains of HLA class Ⅱ (DR) were amplified from tuberculosis patients by PCR. The pMT-HLA-DRB expression vectors that carries the HLA-DR 13 chain and pMT-HLA-DRA-P expression vectors which carries the genes of HLA-DR α chain loaded with Mtb antigen were transfected into S2 cells with the method of calcium phosphate transfection. The expressed Mtb peptide/HLA-DR complexes were primarily identified by the method of cell immunohistochemistry. The cell lines expressing Mtb peptide/HLA-DR complexes were used to screen tetramers of CD4+ TCR by flow cytometry. Results S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface were obtained, two kinds of Mtb specific tetramers of CD4+α/β TCR were screened. Conclusion S2 cell lines expressing Mtb peptide/HLA-DR complexes on the cell surface provide the solid basis of the further research on the TCR tetramers and are helpful for exploring new diagnostic study methods about tuberculosis and developing new vaccines.

Objective Take alkaline ashing method as golden standard to explore the accuracy of chloric acid digestion method in determination of human milk iodine. Methods Sixty one breast milk samples collected in Hexi district of Tianjin was measured by the method for determination of iodine in foodstuff by As3+-Ce4+ catalytic spectrophotometry (referred to as the alkaline ashing method) published in 2008 and the method for determination of iodine in urine by As3+-Ce4+ catalytic spectrophotometry(referred to as acid digestion) published in 1999, respectively. were highly correlated(r = 0.960, t = 26.3, P < 0.01), and the regression equation was (Y) = - 28.1 + 0.808X, in which X was independent variable, that is the results of alkaline ashing method; (Y) was dependent variable, that is the estimated data of chloric acid digestion method. The average difference of the results measured by the two methods was 68.3 μg/L, and the results from chloric acid digestion was 38.9% which lower than that of alkaline samples were diluted by 3,4 and 5-fold and then digested by chloric acid, the liquid clarification rates were 80.3% ashing and chloric acid digestion method were, respectively, 165.4, 110.0 μg/L. Conclusions Compared with alkaline ashing method, the results determined by chloric acid digestion method are significantly lower. It is suggested that there are systemic errors in chloric acid digestion method, which means that alkaline ashing method can not be replaced by the chloric acid digestion method.

Objective To investigate the causes of abnormal decrease in maximum amplitude(MA)of thromboelastog-raphy(TEG)and its effect on prognosis by monitoring the changes of coagulation-related indexes in emergency trauma pa-tients.Methods A total of 319 cases of trauma patients admitted to our hospital from September 2020 to September 2023 were retrospectively analyzed,and the coagulation-related indexes of 0 h and 24 h after admission were observed.According to the MA results,they were divided into normal MA group(>50 mm)and reduced MA group(≤50 mm)to compare the hemoglobin(Hb),platelets count(Plt),activated partial thromboplastin time(APTT),prothrombin time(PT),fibrinogen(Fib),thrombin time(TT),D-dimer(D-D),coagulation reaction time(R),clot formation kinetics(Angle),30 min clot dissolution rate(Ly30),MA,thrombine-antithrombin complex(TAT)and plasminase-α2 plasminase inhibitor complex(PIC).The correlation between MA and fibrinolysis indexes in 319 trauma patients was analyzed.According to whether tranexamic acid(TXA)was used,the reduced MA group was divided into a TXA group and a non-drug group.The differ-ences in the change of the above coagulation-related indexes,mortality rate and changes in blood product dosage were com-pared between the two groups.Results Compared with the normal MA group,Hb,Plt,Fib,diastolic blood pressure and GCS scores decreased,while heart rate,ISS score and mortality increased significantly in the reduced MA group(P<0.05).The R,PT and TT were prolonged significantly(P<0.05),and PIC and D-D increased significantly(P<0.05)in the re-duced MA group.Correlation analysis found that MA had no correlation with Ly30,TAT and APTT,but was correlated with Angle(r=0.803),Plt(r=0.544),Fib(r=0.581),PIC(r=-0.443)and D-D(r=-0.343).Compared with the non-drug group,the change of Angle,MA and FIB in the TXA group increased significantly(P<0.05),while the change of PIC de-creased(P<0.05).Cryoprecipitate and platelet transfusion in the TXA group reduced significantly(P<0.05),and red blood cell transfusion had a decreasing trend,but the difference was not significant(P>0.05).The mortality rate in the TXA group was reduced significantly(P<0.05).Conclusion Hyperfibrinolysis may be an important factor in the abnormal decrease of MA in emergency trauma patients.Treatment with TXA can improve its effect on MA,and reduce the transfusion of blood products and the patient mortality.

OBJECTIVESTo investigate the correlation between MAGE-A1 mRNA expression and genic demethylation in hepatoma cell lines.

METHODSTotal RNA and genomic DNA were prepared from 10 human hepatoma cell lines. MAGE-1 mRNA expression was determined with RT-PCR and the level of genome-wide demethylation was evaluated by enzyme digestion and Southern blot assay. The genomic DNA was digested by HpaII, then the promoter of MAGE-A1 gene was amplified with primers CDS21, EDP4 and CDS20. EDP4 and the PCR products were further hybridized with a probe to detect the methylation in the promoter of the MAGE-A1 gene. HLA-A locus was typed using SSP kit.

RESULTSIn cell lines QGY-7703, SMMC-7721, HLE, BEL-7402, BEL-7404 and BEL-7405, MAGE-A1 mRNA expression was positive and cell differentiation was moderate or low. In cell lines of HepG2 2.2.15, HepG2, QGY-7701 and Huh7, MAGE-A1 mRNA expression was negative and cell differentiation was well or moderate. The level of genomic demethylation in MAGE-A1 mRNA positive cell lines was much higher than that in MAGE-A1 mRNA negative cell lines (t = 2.896, P = 0.02). The methylation analysis showed that methylation in the promoters of MAGE-A1 gene of HepG2 2.2.15, HepG2, QGY-7701 and Huh7 was high, and that methylation in those of SMMC-7721, HLE, BEL-7402, BEL-7404, and BEL-7405 was low.

CONCLUSIONThe results suggest that MAGE-A1 mRNA expression in the human hepatoma cell lines is associated with genic hypomethylation.

Antigens, Neoplasm ; Carcinoma, Hepatocellular ; genetics ; immunology ; metabolism ; DNA Methylation ; Humans ; Liver Neoplasms ; genetics ; immunology ; metabolism ; Melanoma-Specific Antigens ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured

OBJECTIVETo study the responses of peginterferon-alpha 2a antiviral therapy in chronic hepatitis B (CHB) patients.

METHODSOne hundred two CHB patients with their serum ALT values higher than 2x the upper limit of the normal (ULN) were divided into a HBeAg-positive and a HBeAg-negative group. All patients were treated with peginterferon-alpha 2a by subcutaneous injection (180 microgram once weekly). After treatment for 6 months, patients without a defined therapeutic response were dropped from the treatment group; the others completed a 12 month therapy. The sustained response and the antiviral effect of the treatment were assessed at the end of the therapy. To investigate the possible impact factors of the response to peginterferon-alpha 2a, we studied the therapeutic response of patients with different serum ALT levels, inflammation grades of liver histology, stages of fibrosis, and HBV viral load levels.

RESULTS(1) There was no statistical difference of the rates of response at the end of treatment and 6, 12, 18, 24 and 30 months after the cessation of therapy between the HBeAg-positive and the HBeAg-negative groups. (2) In the HBeAg-positive group, the rates of response of patients with serum ALT values>3*ULN were significantly higher than those with serum ALT values less than 3*ULN (x2=4.40). However, no statistical difference of serum ALT levels was found in the HBeAg-negative group. (3) In both HBeAg-positive and HBeAg-negative groups, no difference was revealed in the rates of response among patients with different levels of HBV viral loads. (4) In the HBeAg-positive group, patients with more severe liver inflammation histologically (G3 and G4) had significantly higher response rates than those with milder inflammation (G1 and G2) (x2=4.19), but no similar statistical differences were found in the HBeAg-negative group. Moreover, there was no difference in the rates of response among patients in different stages of liver fibrosis in both HBeAg-positive and HBeAg-negative groups.

CONCLUSIONSSimilar rates of response and sustained virological response to the peginterferon-alpha 2a treatment can be achieved in both HBeAg-positive and HBeAg-negative patients. Hepatic fibrosis is not a predictor of poor therapeutic response. For HBeAg-positive patients, more severe liver inflammation identified with liver biopsies (G3 or G4) and high serum ALT values (more than 3*ULN) can be considered as predictors of a good therapeutic response.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; drug therapy ; pathology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; Young Adult