Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To investigate the state of endemic fluorosis, running status of water improvement project to reduce fluoride in Jiangsu province, and to provide a scientific basis for prevention and control of endemic fluorosis. Methods In 2008, in the ten key counties of endemic fluorosis (zone), in Xuzhou,Lianyungang and Suqian, a stratified sampling method was employed to select 40 diseased villages according to their past water fluoride survey data. All children aged 8 to 12 were examined dental fluorosis, and all adults over 16 years were examined clinical skeletal fluorosis. Thirty per cent of the 40 diseased villages were selected, and 20 adults over the age of 16 in each selected village were examined by X-ray, respectively;50% of the 40 diseased villages were selected, and 30 any time urine samples of children aged 8 to 12 in each diseased village were tested urine fluoride. In each city, select a county, the status of water improvement project to reduce fluoride, water supply capacity and coverage in the county were investigated. Results A total of 3560 children aged 8 to 12 were examined, the detection rate of dental fluorosis was 38.51% (1371/3560), tooth defect rate was 5.34% (190/3560), and dental fluorosis index was 0.8. Seven hundred and eight urine samples were tested, the median urinary fluoride was 1.47 mg/L and the range was 0.08 ~ 10.08 mg/L. Clinical detection of skeletal fluorosis was 21.3% among adults over the age of 16, and X-ray detection rate of skeletal fluorosis was 39.2% (123/314).Investigated a total of 248 facilities of centralized water improvement projects, no funds to run or damaged 49, the water fluoride > 1.0 mg/L was 18. Conclusions Endemic fluorosis in Northern Jiangsu province has not been controlled completely, but has a rising trend, we should further strengthen the supervision of water fluoride reduction.

Osteoarthritis (OA) is a complex chronic progressive disease attacked by biological and mechanical factors and a result from the anabolic and catabolic imbalance in chondrocyte, subchondral bone and extracellular matrix(ECM). Etiology and pathological of OA are not yet entirely clear. The degradation and destruction of collagen II caused by matrix metalloproteinase -13 (MMP-13) is considered the core factor in the occurrence and development of OA. The research of MMP-13 inhibitor provide ideas and methods for the treatment of OA. In this article,the role and determination of MMP-13 in OA and the development prospect of MMP-13 inhibitor in the treatment of OA research progress were reviewed.
Animals ; Collagen ; metabolism ; Humans ; Matrix Metalloproteinase 13 ; analysis ; physiology ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Osteoarthritis ; drug therapy ; etiology

OBJECTIVEThis paper is to explore the relationships between the occurrence of structural birth defects and environmental risk factors on birth defects.

METHODSA survey was conducted by using a population-based case control study. A total number of 388 structural birth defects, which were detected after 28 weeks' gestational age to 7th day after birth in various hospitals were recruited in Wuxi from 2002 to 2004. The control group consisted of 1607 normal live births born in the same period. Multiple logistic regression model was used to analyze relationships between environmental risk factors and occurrence of structural birth defects after controlling women's education level and childbearing age.

RESULTSAmong those factors which affected birth defects, the exposure to toxic substances in working environment was the highest risk factor, with OR value as 5.37 (95 % CI : 3.60-7.99). Suffering from reproductive tract infections, high fever during first trimester gestation, and taking potential teratogenic drugs such as hypnotics and anti-convulsion drug agents, might significantly increase the risk of structural birth defects, with OR values (95% CI) as 3.38 (1.33-8.56), 3.57 (1.73-7.37) and 2.75 (1.57-4.83) respectively. However, the correlation between oral contraceptives intake within six months before pregnancy and risk of birth defects had no statistical significance. In addition, pollution of the residential environment, staying up all night and raising pets at home, had relatively lower association with birth defects, with OR values (95% CI) as 2.28 (1.75-2.98), 1.96 (1.21-3.18) and 2.50 (1.66-3.76) respectively, yet with a higher proportion of exposure to those factors in pregnant women. There was a significant increase in occurrence risks of birth defects with increase in the number of exposure to environmental risk factors. OR values for having one, two, three kinds of environmental risk factors were 1.60, 4.32 and 10.23 respectively.

CONCLUSIONThe common structural birth defects were affected by a number of environmental risk factors. There was also a dose-response relationship between the number of environmental risk factors and occurrence risks of structural birth defects noticed in our study.

Air Pollution ; adverse effects ; Case-Control Studies ; Congenital Abnormalities ; etiology ; Female ; Humans ; Logistic Models ; Maternal Exposure ; adverse effects ; Pregnancy ; Risk Assessment ; Risk Factors

Objective To study the effect of physical activities and dietary intervention on metabolic syndrome (MS) in primary school students.Methods Either one-year physical activities (PA) or dietary intervention was conducted in Grade 1-5 children from eight primary schools in Haidian district,Beijing.A ‘happy 10 minutes' program was held in the PA group,while the dietary group receiving nutrition lectures.Baseline and post-intervention data on height,weight,waist circumference,serum lipids,glucose,and blood pressure were collected.Results The prevalence of MS at baseline was 9.0％.After intervention programs were carried out,improvements on triglyceride,high density lipoprotein,fasting blood glucose,diastolic blood pressure and waist circumference in the dietary group or PA group were seen,with the dietary group showed better effects than the PA group.The prevalence of MS decreased from 10.4％ to 4.6％ in the dietary group with statistically significant difference,while it increased in both the PA group and the control group.Girls showed better effects in the dietary group.Older students showed better effects than the younger students in the PA group.In non-overweight and overweight students of the dietary intervention group,the MS related components were significantly improved.Conclusion The prevalence of metabolic syndrome was relatively high in the primary school students in Beijing.Education on nutrition could reduce the prevalence and improve the related components,which seemed to be more effective than in the PA intervention.Sex,age and nutritional status were the confounding factors for intervention programs.

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans

The low sensitivity of microbial fuel cell ( MFC)-biosensor is one of the bottlenecks in its practical application. To investigate the effect of anode electrode modified with carbon nanomaterials on the sensitivity of water toxicity detection of MFC-based biosensor, graphite felts ( GF ) were modified using two carbon nanomaterials of multi-walled carbon nanotubes ( MWNT ) and conductive carbon black ( GCB ) . MFC biosensors were started up with the anode electrodes, and the results showed that resistance of the GCB and MWNT-modified electrode was smaller than that of unmodified electrode, and the order of MFC power output was GCB/GF-MFC (2. 63 W/m2)>MWNT/GF-MFC (2. 56 W/m2)>GF-MFC (2. 09 W/m2). Then, 3, 5-dichlorophenol poison ( DCP) was used as a model toxicant in toxicity test, the order of toxicity inhibition ratios of 10 mg/L DCP to three MFC biosensor was MWNT/GF-MFC (31. 8% )>GCB/GF-MFC (26. 3% )>GF-MFC (20. 1% ). The sensitivity for toxicity detection by MFC biosensors with anode electrode modified with carbon nanomaterials was improved, and MWNT/GF-MFC had the highest sensitivity. The result of the study may promote the application of MFC biosensor in water pollution monitoring.

OBJECTIVETo investigate the effect of intraluminal administration of ulinastatin (a protease inhibitor) in the intestine on intestinal inflammation in rats with hemorrhagic shock.

METHODSTwenty-eight Wistar rats were randomized into control group (A), intestinal saline perfusion group (B), ulinastatin intestinal perfusion group (C), and intravenous ulinastatin injection group (D) (n=7). The mean arterial blood pressure (MAP) and survival time of the rats were recorded. The changes in human polymorphonuclear cell (PMN) CD11b expression were detected by flow cytometry. The leukocyte count was recorded at different time points after the treatment, and the pathology of the intestinal mucosa was observed comparatively.

RESULTSGroups C and D showed significantly slower reduction of the MAP than groups A and B after hemorrhagic shock (P<0.05). The survival time of the rats was the longest in group C (P<0.05). CD11b expression increased gradually during hemorrhagic shock in all the groups, but the expression level was the lowest in group C (P<0.05). Hemorrhagic shock caused a reduction in leukocyte counts, which remained the highest in group C (P<0.05). Group C also showed the least intestinal pathology among the 4 groups.

CONCLUSIONIntestinal perfusion of ulinastatin can lower the reduction rate of MAP, attenuate plasma activation and intestinal inflammation, and prolong the survival of rats with hemorrhagic shock. These results indicate an important role of protease in intestinal inflammation during hemorrhagic shock.

Animals ; Arterial Pressure ; Disease Models, Animal ; Glycoproteins ; administration & dosage ; pharmacology ; Inflammation ; enzymology ; metabolism ; Intestines ; enzymology ; metabolism ; Plasma ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; blood ; enzymology ; metabolism ; Trypsin Inhibitors ; administration & dosage ; pharmacology

Objectives: To evaluate the clinical characteristics and prognosis of high risk cytogenetic abnormalities (HRCA) and various combinations of cytogenetic abnormality in patients with newly-diagnosed multiple myeloma (NDMM) . Methods: This retrospective study collected 182 NDMM patients in the First Affiliated Hospital of Jilin University between Nov. 2009 and May 2018. HRCA included 1q+, del (17p) , t (4;14) , and t (14;16) detected by FISH, and non-HRCA included del (13q) , t (11;14) detected by FISH. The clinical characteristics among three groups, including cases who carrying a single HRCA, 1 HRCA in combination with non-HRCA and cases carrying two or more HRCAs (double/triple-hit) were observed. Kaplan-Meier curve was used to analyze both progression-free survival (PFS) and overall survival (OS) for the three groups. Results: The survivals of patients with 1 HRCA in combination with non-HRCA were similar to those with two or more HRCAs (double/triple-hit) , the median PFS (mPFS) was 19.1 m vs 12.1 m (P=0.248) and median OS (mOS) was 29.6 m vs 29.3 m (P=0.774) . Furthermore, the prognosis of these two groups were both inferior to patients with a single HRCA, respectively. (mPFS: 32.2 m, P=0.040, P=0.001; mOS: 42.3 m, P=0.021, P=0.041) . Strikingly, both the mPFS and the mOS of patients with 1 HRCA in combination with non-HRCA (regardless of high risk or not) were significantly shorter than that of cases with a single HRCA (mPFS: 15.1 m vs 32.2 m, HR=2.126, 95%CI 1.176-3.843, P=0.005; mOS: 29.3 m vs 42.3 m, HR=1.442, 95%CI 0.705-2.950, P=0.011) . Conclusion: It is of prognostic significance value for detecting double/triple-hit based on FISH cytogenetics in NDMM.
Chromosome Aberrations ; Chromosome Disorders ; Cytogenetic Analysis ; Humans ; Multiple Myeloma ; Prognosis ; Retrospective Studies

Objective: To evaluate the prognostic value of kinetic changes in minimal residual disease (MRD) status, as well as its relationship with risk stratification, therapeutic response and treatment in patients with newly-diagnosed multiple myeloma (MM) . Methods: A total of 135 patients with newly-diagnosed MM were screened, and 105 patients who achieved VGPR or more as the best responses were included into this study. The MRD status was determined by multiparameter flow cytometry (MFC) at multiple intervals after two cycles of treatment until clinical relapse, death, or last follow-up. The statistical methods included Kaplan-Meier analysis, Cox regression, etc. Results: ①In all 135 patients, 57.8% (78/135) patients achieved MRD negativity (MRD(-)) after treatment. In 105 patients who achieved VGPR and thus included in this study, the MRD(-) rate was 72.4% (76/105) , with a median interval of 3 months from starting treatment to achievement of MRD(-) status. ②The 2-year PFS rate of patients with MRD(-) status was significantly higher than that of MRD(+) status (62.2% vs 41.3%, P=0.001) , while MRD persistence (MRD(+)) was an independent factor for poor prognosis (multivariate analysis for PFS: P=0.044, HR=3.039, 95%CI 1.029-8.974) . ③Loss of MRD(-) status (i.e., MRD reappearance) showed inferior outcomes compared with MRD sustained negative ones, the PFS was 18 months versus not reach (P<0.001) and the OS was not reach for both (P=0.002) . ④The 2-year PFS and OS rates of patients with duration of MRD(-)status≥12 months were significantly higher than those of the control group (PFS: 77.7% vs 36.7%, P<0.001; OS: 96.4% vs 57.9%, P<0.001 respectively) . Duration of MRD(-) status was associated with a marked reduction in risk of relapse or death (univariate analysis for PFS: P<0.001, HR=0.865, 95%CI 0.815-0.918; for OS: P=0.001, HR=0.850, 95%CI 0.741-0.915 respectively) . ⑤Moreover, even in patients carrying high-risk cytogenetic abnormalities (CA) or ineligible for ASCT, MRD negativity remained its prognostic value to predict PFS (high-risk CA medianPFS: not reach vs 19 months, P=0.006; ineligible for ASCT medianPFS: not reach vs 25 months, P=0.052 respectively) . ⑥Last, treatment with the bortezomib-based regimens contributed to prolonged MRD(-) duration (median MRD(-) duratio: 25 months vs 10 months, P=0.034) . Conclusion: Our findings supported MRD(+) status as an independent poor prognostic factor in MM patients, which implicated that duration of MRD(-) status also played a significant role in evaluation of prognosis, while loss of MRD(-)status might serve as an early biomarker for relapse. Therefore, monitoring of MRD kinetics might more precisely predict prognosis, as well as guide treatment decision, especially for when to start retreatment in relapsed patients.
Bortezomib/therapeutic use* ; Humans ; Multiple Myeloma/therapy* ; Neoplasm Recurrence, Local ; Neoplasm, Residual/diagnosis* ; Prognosis ; Risk Assessment ; Treatment Outcome