Viable but non-culturable state in bacterial cells as received uncultured microorganism has be- come the fundamental research issues in medical microbiology, epidemiology, general microbial ecology and sanitation quarantine since it was put forward in 1982. When in this state, bacterium are no longer able to grow and form colonies on conventional culture media, but maintain their patheogenicity, may become latent infection escaping detection to threaten enviroment and human security. With the development of modern molecular biology technique and metagenomics, it provide new research method and opportunity for uncul- tured microorganism in the environment. Recently, with the application of the metagenome technique, ge- netic fingerprinting technical etc, it becomes more and more popular. Simultaneously, along with this state become maturity in the laboratory. It provides a new research route for development and exploitation the uncultured microorganism.

Objective To study on the effectiveness of endoscopic ultrasonography(EUS)in detec- ting insulinoma preoperatively.Methods Fifteen patients with clinical and biochemical signs of insulinoma were examined by EUS using a radial-scanning ultrasound endoscope and abdominal ultrasonography,CT, DSA prior to surgery.The outcome was evaluated on the basis of surgery and examination of the resected specimens.Results Fifteen patients with 16 lesions of insulinoma were identified by surgery and pathology. The aceuraey of diagnosis with EUS was 13/15(86.7%),and that with B-US,CT,DSA was 3/15(20%), 5/15(33.3%),9/14(64.3%)respectively.In the 14 lesions identified by EUS,10 lesions were depicted to be hypoechogenic,1 lesion was isoechogenic and 3 lesions were hyperechogenie.All 14 lesions were well demarcated and surrounded by normal pancreatic tissue.The minimum size of the lesion visualized by EUS was 0.5cm.Ten lesions were correctly detected by EUS with size of 0.5～2.0cm.EUS missed diagnosis in 2 lesions not for their small size.EUS falsely indicated a 10mm lesion from two lesions inside the head of pancreas.One lesion outside the pancreatic tail and one lesion in the pancreatic head were missed by EUS in another case.Conclusion EUS is superior in assessing the location of pancreatic insulinoma than other ima- ging methods such as B-US,CT,DSA.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

Objective To know the condition and feature of water-born endemic arsenism in Shaanxi province, and to provide scientific evidences for prevention and treatment of the disease. Methods In 2009, 8 villages of Duhe, Miaogou, Sanhe, Caiyuan in Shanyang county and Shuanglong, Guangming, Donghekou in Zhen'an county and Guojiawan in Mian county as monitoring sites from both the found water-born arsenic poisoning areas and potential ones of drinking water arsenic ＞ 0.15 mg/L were selected. The disease conditions of residents who lived for more than six months of continuous residence, were investigated according to "Endemic Arsenism Diagnostic Criteria" (WS/T 211-2001 ) and arsenic level in the drinking water were determined. Results Seven thousand six hundred and ninety-three individuals were investigated and 612 cases of arsenism were diagnosed in the 8 villages, the prevalence rate was 7.96%(612/7693). Mild patients accounted for 33.17%(203/612), moderate patients accounted for 50.16%(307/612), and severe patients accounted for 16.67%(102/612). The detection rate for male was 9.99%(408/4085), for female was 5.65%(204/3608), the former was higher than the latter(x2 = 49.138,P ＜ 0.01 ). The older the age was, the higher the rate was(x2 = 770.708, P ＜ 0.01 ). Main clinical manifestation of the patients was depigmentation and pigmentation in skin. Patients with one symptom accounted for 43.30% (265/612), with two symptoms accounted for 56.70% (347/612). Three hundred and seventy drinking water samples were detected, the median of arsenism content was 0.005 mg/L, ranged 0 - 1.660 mg/L. The over-limit rate of arsenic level in the drinking water were 11.62% (43/370). Conclusion Water-born endemic arsenism is mildly prevailing in Shaanxi province, however, great attention should be given to control and monitoring of the disease.

Objective To investigate the current situation of endemic fluorosis associated with drinking water in Shaanxi province in 2008, providing prevention and control work with scientific evidence. Methods According to data of drinking water endemic fluorosis in Shaanxi, 15 counties in Yulin, Weinan and Xianyang city were divided into mild, moderate and severe diseased areas according to fuoride level in the drinking water or the fluorosis severity, and 12,39,45 endemic fluorosis villages were selected respectively from each area. Dental fluorosis for all the children aged 8 - 12 years old and clinical skeletal fluorosis for the people over the age of 16 were examined. In mild, moderate and severe areas, 30% of the survey villages received X-ray to examine skeletal fuorosis. Five water samples were collected from villages with unimproved water respectively from the east, the west, the south, the north, the middle; if there was less than 5 that could be sampled, then all were collected, moreover 3 peripheral and 1 central water samples were collected from villages with improved water. Also in mild, moderate and severe illness areas, 50% of the surveyed villages were selected; in each village, 30 urine samples of the children aged 8 - 12 were collected, 6 portions in each age group. If Less than 30 children in all the age groups or less than 6 in each age group, all their urine samples were collected to determine fluoride. Results Totally 3652 children aged 8 - 12 years old were examined and 1930 cases of dental fluorosis were diagnosed, the rate was 52.85% and the index was 1.22. The rates in Dinghian, Jingbian and Pucheng village were higher, being 90.43% (170/188) ,82.89%(126/152),80.65% (325/403) respectively. A total of 40 543 adults over the age of 16 were examined and 5935 cases of skeletal fluorosis were diagnosed, in a rate of 14.64%. The rates of the mild, moderate, severe skeletal fluorosis were respectively 9.17%(3717/40 543),4.36%(1769/40 543),1.11%(449/40 543).All 706 adults were taken X-ray films.and 280 cases of positive change were diagnosed,the positive rate was 39.66%. The rates of the mild,moderate,severe change were respectively 17.28%(122/706),17.00%(120/706),5.38%(38/706).All 427 water samples were collected,the fluoride content waS(2.22±1.41)ms/L,and the abnormal rate was 85.01%(363/427).All 1393 portions of urine sample were collected in children and the median of fuoride was 2.37 mg/L Conclusions The endemic fluorosis associated with drinking water is still comparatively serious in Shaanxi.The preventive measures for improving water must be improved.

ObjectiveTo investigate the current situation of coal-burning borne endemic fluorosis in the infected area,the usage of defluoridation stove and the formation of related behavior; to evaluate the control effect,so as to provide a theoretical basis for superiors' policy-making.MethodsNine villages in three counties of Ankang city and Hanzhong city Shanxi province were monitored in 2010.Choose five disease infected villages with oven changed in the three monitoring counties,and 10 families were randomly selected in each of the nine monitoring points and every chosen village,and a survey on the stove using and related behavior changes was carried out.By using Dean method,the dental fluorosis of every child aged 8 - 12 in the monitoring villages was examined.Meanwhile 10 copies of urine samples were randomly collected in each age group in the above-mentioned villages.The content of fluoride in urine was determined by F-ion selective electrode(WS/T 30-1996) and evaluated according to “The Normal Concentration of Urinary Fluoride of Population” (WS/T 256-2005).Twenty men over 25years old in the monitoring counties were randomly selected in each age group to be examined skeletal fluorosis by X-rays,and diagnosis was based on “The Endemic Skeletal Fluorosis Standard”(WS 192-2007).ResultsThe proper utilization rate and the qualified households of iron stoves and improved stoves,the correct drying rate and the correct storage rate of peppers and coms were all low.The washing rate of corns and peppers before eating was higher than 95％.Of the 350 children being examined of dental fluorosis in the 9 monitoring villages,the detection rate of dental fluorosis was 59.71％ (209/350).A total of 871 adults over the age of 25 were examined,and the detection rate of skeletal fluorosis was 17.22％ (150/871).A total of 350 copies of children urine samples were tested and the fluoride content were between 0.07 - 2.02 mg/L,the geometric mean value of fluoride content was 0.66 mg/L.ConclusionsThe coal-burning borne endemic fluorosis in Shanxi is in a growth phase,many defluoridation stoves are out of order,and the formation of healthy behaviors is low.To strengthen the disease surveillance,health education and management of defluoridation stoves are keys to prevent coal-burning borne endemic fluorosis in the future.

Objective To investigate the running conditions of the water improvement projects and the role of these projects in reducing fluoride in drinking-water type of fluorosis in Shaanxi province, and provide a scientific basis for prevention and control of the disease. Methods Forty-eight villages of 16 counties in Xi'an, Baoji, and Yulin cities of Shaanxi province were monitored in 2009. Five water samples were collected randomly in water unimproved monitoring villages by the position of east, west, south, north, and center parts. In water improved monitoring villages, 3 tap water and one source water samples were collected. Water fluoride was tested using fluoride ion selective electrode method according to the "Standard Testing Methods for Drinking Water" (GB/T 5750-2006). All school children aged 8 to 12 in monitored villages were examined their dental fluorosis using Dean criteria. All people over 16 years old were examined clinical skeletal fluorosis, and 30% of the project counties were randomly selected, then randomly selected one village among these counties, clinically diagnosed patients with skeletal fluorosis were examined again by X-ray using "Diagnostic Criteria of Endemic Skeletal Fluorosis"(WS 192-2007). Urine samples of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected, urinary fluoride was tested according to "the Determination of Urinary Fluoride by Ion Selective Electrode Method" (WS/T 89-19%). Results Of the 22 water improvement projects that in normal operation, fluoride level of 8 source waters exceeded the standard, accounting for 36.36%(8/22), and projects scrapped 5. Two hundred and two water samples were tested. In water improved historical diseased areas, the median of water fluoride of source water and tap water were 0.72,0.62 mg/L, respectively, and the average rate of water fluoride exceeded the standard ere 36.36%(8/22) and 31.94%(23/72), respectively. In water unimproved historical diseased areas, the median of water fluoride was 1.00 mg/L, and the average rate of water fluoride exceeded the standard was 39.81%(43/108). Detection rate of dental fluorosis among children aged 8 to 12 was 16.06% (367/2285), dental fluorosis index was 0.30, and the prevalence was negative. Detection rate of clinical skeletal fluorosis among adults over 16 years old was 5.09%(1542/30 272), a totally of 198 people had X-ray film taken, positive 68, the positive detection rate was 34.34%(68/198). One thousand and fifty-one copies of children's urine samples were tested, geometric mean of urinary fluoride was 0.95 mg/L; nine hundred and fourteen copies of adults urine samples were tested, geometric mean of urinary fluoride was 1.16 mg/L Conclusions Drinking-water type of fluorosis affects a large area in Shaanxi province, the disease is still serious, and the task of prevention remains very arduous. Further intensify the water improvement project in diseased areas, and strengthen disease monitoring, health education and water improvement project management is the key to prevention and control of endemic fluorosis.

OBJECTIVETo explore the influence of arsenic pollution caused by coal-burning on methylation (promoter and exon 5) and mutation (exon 5) of human p53 gene, and to analyze the relationship between methylation, mutation and arsenism.

METHODSAccording to the diagnostic criteria of endemic arsenism, 112 patients with arsenism (including 38 mild cases, 43 moderate cases and 31 severe cases) were selected in the areas with endemic arsenism from Xingren, Guizhou province. Among the subjects, 43 cases were diagnosed by dermatopathological methods, and they were divided into non-cancerous group (24 cases) and cancerous group (19 cases). 90 controls were selected from the non-arsenic polluted areas. Under the principle of informed consent, blood samples were collected from individuals. The methylation of p53 gene in promoter region and exon 5 were detected by extinction enzyme-PCR, the mutation of p53 gene (exon 5) was detected by PCR-SSCP, PCR products cloning and sequencing technology.

RESULTSThe positive rates of methylation of p53 gene in promoter region were 13.16% (5/38), 27.91% (12/43) and 45.16% (14/31) respectively among mild, moderate and severe arsenism group, which were obviously higher than the rates in the control group (1.11% (1/90), χ² values were 8.679, 23.690, 41.199, respectively, both P values < 0.017). The positive rates of methylation of p53 gene were 25.00% (6/24) and 63.16% (12/19) in non-cancerous and cancerous group respectively, which were obviously higher than those in the control group (1.11% (1/90), χ² values were 18.762, 57.497, respectively, both P values < 0.025). The positive rates of methylation of p53 gene (exon 5) were 55.26% (21/38), 51.16% (22/43) and 48.39% (15/31) respectively among mild, moderate and severe arsenism group, which were obviously lower than the rates in the control group (88.88% (80/90), χ² values were 18.151, 23.168, 22.420, respectively, both P values < 0.017). The positive rates of methylation of p53 gene (exon 5) were 54.17% (13/24) and 42.11% (8/19) in non-cancerous and cancerous group respectively, which were obviously lower than those in the control group (88.88% (80/90), χ² values were 15.201, 22.075, respectively, both P values < 0.025). The mutation rates of p53 gene (exon 5) were respectively 5.26% (2/38), 16.28% (7/43) and 25.81% (8/31) among mild, moderate and severe arsenism group; while the results in moderate and severe arsenism group were obviously higher than in the control group (0.00%, χ² values were 15.465, 24.870, respectively, both P values < 0.017). The positive rate of mutation of p53 gene (exon 5) were respectively 16.67% (4/24) and 31.58% (6/19) in non-cancerous and cancerous group, which were obviously higher than it in the control group (0.00%, χ² values were 15.545, 30.077, both P values < 0.025). The hypermethylation of p53 gene in promoter region was related with the mutation of p53 gene (exon 5) (coefficient of association was 0.294, P value < 0.05); and the hypomethylation of p53 gene (exon 5) was related with the its mutation (coefficient of association was 0.410, P value < 0.05).

CONCLUSIONArsenic pollution caused by coal-burning can cause the hypermethylation of p53 gene in promoter region, hypomethylation and mutation of p53 gene (exon 5), and the changes of methylation of p53 gene are related with its mutation and might be one of the important etiological factors of arsenic pathogenicity or carcinogenesis.

Adult ; Arsenic Poisoning ; etiology ; genetics ; Case-Control Studies ; Coal ; adverse effects ; DNA Methylation ; Environmental Pollution ; adverse effects ; Female ; Genes, p53 ; Humans ; Male ; Middle Aged ; Mutation ; Promoter Regions, Genetic ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo study the rearrangement of immunoglobulin (Ig) heavy chain variable region (V(H)) genes in human neonates with different gestational ages (GA).

METHODSPeripheral blood from the neonates with GA of 27 weeks (4 cases), 28-32 weeks (9 cases), 33-36 weeks (12 cases), and 37-42 weeks (13 cases) was collected. RT-PCR was used to amplify the Ig V(H) gene, and the PCR products were separated by electrophoresis and analyzed using 6% denaturing PAGE gel.

RESULTSAll Ig V(H) family genes had several rearranged genes in each GA group, and the neonates with different GA showed no significant difference in the median molecular weight for each rearranged Ig V(H) family gene.

CONCLUSIONThe neonates with GA of 27-42 weeks exhibit diversity in Ig V(H) gene rearrangement, and for the same Ig V(H) family, the median length of the arranged Ig V(H) genes is independent of the gestational age.

Gene Rearrangement ; Genes, Immunoglobulin Heavy Chain ; Gestational Age ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Infant, Newborn ; Multigene Family ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo investigate the association between non alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MS).

METHODSA cross-sectional multiple-stage stratified survey was performed. A total of 2190 civil servants of Chongqing city were invited to participate in the survey covering physical examination, serum biochemistry-profile and ultrasonographic examination of liver.

RESULTSOf 2176 valid questionnaires, altogether 455 cases were diagnosed as NAFLD and 231 individuals were diagnosed as MS. The prevalence of obesity, hyperglycemia, blood lipid disturbance, primary hypertension, NAFLD and MS was 38.3%, 5.5%, 31.7%, 29.9%, 20.9% and 10.6% respectively, which was increased along with aging (chi2 = 31.775, P = 0.000; chi2 = 25.985, P = 0.000; chi2 = 44.818, P = 0.000; chi2 =149.802, P = 0.000; chi2 = 61.302, P = 0.000; chi2 = 43.508, P = 0.000 a partly). The prevalence of obesity, hyperglycemia, dyslipidemia, primary hypertension, metabolic syndrome was significantly higher than those in control group (chi2 = 384.554, P = 0.000; chi2 = 25.597, P = 0.000; chi2 = 370.849, P = 0.000; chi2 = 40.252, P = 0.000; chi2 = 215.077, P = 0.000 separately), and the level of body mass index (BMI), fasting plasma glucose (FPG), triglyceride (TG), systolic blood pressure (SBP), diastolic blood pressure (DBP) in NAFLD group was remarkably higher than those in control group (t = 26.308, P = 0.000; t = 6.055, P = 0.000; t = 15.980, P = 0.000; t = 10.550, P = 0.000; t = 13.628, P = 0.000 respectively), while the level of high-density lipoprotein cholesterol (HDL-C) was on the opposite (t = 20.067, P = 0.000). Compared with the control group, odds risk for NAFLD was 22.82 folds (95% CI: 12.64-41.19) in obesity, 20.97 folds (95% CI: 11.21-39.24) in hyperglycemia, 24.40 folds (95% CI: 13.51-44.07) in dyslipidemia, 15.73 folds (95% CI: 8.66-28.60) in primary hypertension, while the risk for NAFLD was the highest in MS (OR = 31.06, 95% CI: 17.12-56.35). There were simple or multiple metabolic disorders in 455 individuals diagnosed as NAFLD, and 21 case (4.6%) with obesity, hyperglycemia, dyslipidemia and primary hypertension.

CONCLUSIONNAFLD is closely related with MS, which may be considered as a feature of MS.

Adult ; China ; epidemiology ; Fatty Liver ; epidemiology ; Female ; Humans ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Prevalence