ObjectiveTo study the relationship between Borna disease virus (BDV) and the human viral encephalitis.MethodsThe P24 fragment of BDV RNA in peripheral blood mononuclear cells (PBMC) from 59 patients with viral encephalitis diagnosed clinically, and 112 healthy donors were examined by fluorescence quantitative nested reverse transcriptase polymerase chain reaction (FQ-nRT-PCR).ResultsThere were 3 positive of BDV P24 fragment in 59 patients with viral encephalitis, and no positive in blood donors. The positive rate of BDV p24 in PBMC in viral encephalitis (5.08%) was higher significantly than that in blood donors (P

Background:Clinical and epidemiological studies revealed that estrogen replacement therapy was associated with a significant reduction in risk of colorectal cancer in postmenopausal women.In our previous studies,estrogen increased the expression of mismatch repair (MMR)gene MLH1 in colonic cancer cells,and re-expression of MLH1 in MLH-deficient colonic cancer cells significantly increased the estrogen-induced apoptosis.Aims:To investigate the signaling pathway implicated in the MLH1-mediated apoptosis in colonic cancer cells induced by estrogen and the roles of p53 and its related genes in this apoptotic pathway.Methods:Plasmid containing wild type human MLH1 (hMLH1)full length cDNA was transfected into MLH1-deficient human colonic cancer cell line HCT116.By using HCT116 cells transfected with empty plasmid as controls,the apoptotic DNA ladder was determined by electrophoresis and the expressions of p53 and other apoptosis-related proteins were assessed by Western blotting under the condition with or without estrogen stimulation. Results:17β-estradiol (E2 )at the concentration of 10 -8 mol/L induced significant apoptosis in HCT116 cells transfected with hMLH1.In HCT116 cells transfected with hMLH1 and stimulated with E2 (group D),the protein expressions of caspase-3,caspase-9,p53,Bax and cytoplasmic cytochrome C increased significantly when compared with HCT116 cells stimulated with E2 only (group B);expressions of the abovementioned proteins were also higher in group D than in group C (transfected with hMLH1 only).Conclusions:MMR gene MLH1 is involved in estrogen-induced apoptosis of human colonic cancer cell line HCT116 by activating p53 signaling and mitochondrial apoptotic pathway.

Objective To explore the therapeutic value of modified endoscopic submucosal dissection with ligation (ESD-L) for small tumors originated from gastric muscularis propria.Methods Total of 60 patients with tumors originated from gastric muscularis propria,which was confirmed by endoscopic ultrasonography and smaller than 12 mm and were recruited to the present study.The conventional ESD technique was used to dissect the tumor to the depth of muscularis propria.Then the bottom of the tumor and the beneath muscularis propria were ligated fully with a nylon loop.Further dissection was applied till the whole tumor was isolated.Results All 60 lesions were dissected completely with perforation occurred in 10 cases,which were managed successfully with metal hemoclip.Pathologic diagnosis was obtained in all lesions,and no recurrence was found during the follow-up.Conclusion Modified technique of ESD-L is effective for complete resection of small tumors originated from gastric muscularis propria,which can also decrease the risk of stomach perforation.

Background Evidences, from recent studies, suggested that Borna disease virus (BDV) infection might be associated with human neuropsychosis, especially psychiatric disorders including depressive disorder(DD). However, controversy existed about the association between BDV infection and pathogenesis of DD. This study was to explore further whether the infection of Borna disease virus (BDV) is associated with the pathogenesis of depressive disorder (DD).Methods The p 24 fragment of BDV RNA in peripheral blood mononuclear cells (PBMCs) from 60DD patients and 120 healthy volunteers was detected by nested reverse transcriptase polymerase chain reaction (nRT-PCR) combined with fluorescence quantitative polymerase chain reaction (FQPCR). Positive products were cloned and sequenced before being compared with Strain V and strain He/80, from humans and animals.Results The positive rate (5%, 3/60) of BDV p 24 in PBMCs from the DD patients was significantly higher than that (0%, 0/120) from healthy volunteers ( P＜0. 05). The gene sequence for the positive products showed BDV p 24 in PBMCs from DD patients in Chongqing was most homophylic with H1766 strain detected from iii horses (97.68%), with 2 situs mutations (nt 1675 T→C, nt 1678 C→T), and also similar to the standard strain V(96. 51%)and He/80(95.35 %), with basic exchanges limited to T- C and A→G.Conclusions There was BDV infection in the DD patients in China, which indicated that the pathogenesis of DD in human beings in Chongqing might be associated with the infection of BDV.

[Objective] To refer a geographical distribution rule of alpha-L-fucosidase (AFU) reference values for the health adults in China via exploring its spatial distribution trend and its correlation with geographical factors.[Methods] A total of 6564 samples of AFU reference values from 66 administrative units in the years 2004-2015 were collected,male and female of which were 3701 cases (56.4％) and 2863 cases (46.3％).A research concerning AFU reference values in whole country were calculated using methods of information content and ridge regression.[Results] AFU reference values for Chinese healthy adults were influenced by geographical factors and presented autocorrelation,and it showed eastern and northern areas were highery than western and southern areas.[Conclusions] AFU reference values have a spatial variation and the regional disparities should be considered in practice.

Objective To observe the effects of MSH2 gene re expression on estrogen-induced apoptosis of colon cancer cells LOVO,and to explore its mechanisms.Methods According to different plasmid and whether with estradiol intervention,colon cancer LOVO cells were divided into empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,estrogen receptor (ER) β with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group,and received corresponding treatment.The expression of MSH2,ERβ protein and apoptosis related caspase 3 protein were detected by Western blotting.Cell viability was measured by cell counting kit-8.Cell DNA fragments of each group were isolated with apoptosis DNA fragments isolation kit.And the DNA ladder was observed.The rate of apoptosis was detected by flow cytometer.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between the two groups.Results After transfection,the expression of the MSH2 and ERβ at protein level in LOVO cells significantly increased and neither of their expression was effected by estradiol.The expression levels of caspase 3 cleavaged active fragments of ERβ with estradiol group and ERβ with MSH2 and ethanol group were higher than other groups,and there was no significant difference between these two groups.The LOVO cell viability of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 1.72 ±0.25,1.74 ± 0.31,1.77 ± 0.35,1.74±0.33,1.70±0.34,1.02±0.48,1.71±0.31 and 1.07±0.18,respectively,and the differences between the groups were statistically significant (F=3.791,P＜0.05).Among them,the LOVO cell viability of ERβ with estradiol group was lower than that of ERβ with ethanol group,accordingly,that of ERβ with MSH2 and estradiol group was lower than that of ERβ with MSH2 and ethanol group,that of ERβ with estradiol group was lower than that of empty plasmid with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t=3.158,3.075,3.648,3.253,all P＜0.05).DNA ladder formed from DNA fragments of apoptosis cells was seen in ERβ with estradiol group and ERβ with MSH2 and estradiol group.The apoptosis rate of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 7.86±0.19,7.87±0.39,8.39±1.02,9.05±1.54,7.54±0.99,19.77±2.35,7.76±1.32 and 19.30±1.75,respectively,and the differences between groups were statistically significant (F=45.436,P＜0.05).Among them,the apoptosis rate of ERβ with ethanol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and ethanol group was lower than that of ERβ with MSH2 and estradiol group,that of empty plasmid with estradiol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t =8.260,9.133,8.596,7.617,all P＜ 0.05).Conclusions Estrogen may promote colon cancer cell apoptosis through ERβ pathway.The process of apoptosis maybe related with caspase protein,MSH2 may not be involved in the regulation of this signal pathway.

Objective To investigate the reference intervals of mean corpuscular volume (MCV) ,mean corpuscular hemoglo‐bin(MCH) and mean corpuscular hemoglobin concentration (MCHC) examined by the MindrayBC‐6800 hematological analyzer to establish the reference intervals suitable for our laboratory .Methods According to the method recommended by the NCCLS C28‐A3 ,600 healthy adult individuals were selected as the reference individuals .MCV ,MCH and MCHC levels were determined by the MindrayBC‐6800 hematological analyzer for constructing the reference intervals ;other 150 healthy persons undergoing the physical examination were selected and their MCH ,MCV and MCHC detection results were collected for verifying the established reference intervals .Results The detection results of MCV ,MCH and MCHC in healthy adults showed a normal distribution ,MCV had sta‐tistical difference among different age periods (P<0 .05);the reference intervals :82 .278 -94 .242 fL for young adults ,83 .032-94 .608 fL for the middle‐aged persons and 83 .137-96 .343 fL for the elderly .MCH had statistical differences between different se‐xes and among different age periods ;the reference intervals :27 .785-32 .415 pg for male young adults ,28 .324-32 .456 pg for male middle‐aged persons and 28 .274-32 .966 pg for male elderly ;27 .367-31 .973 pg for female young adults ,27 .445-32 .215 pg for female middle‐aged persons and 27 .532 -32 .468 pg for female elderly .MCHC had statistical difference between different sexes (P<0 .05) ;the reference intervals :328 .611-352 .810 g/L for male and 323 .771-348 .750 g/L for female .In 150 individuals un‐dergoing the physical examination ,the proportion of individuals locating at the outside of reference interval was less than 10 .0% , therefore the newly established intervals were suitable for this laboratory .Conclusion The sex difference or/and age differences of MCV ,MCH and MCHC exist among adult populations .So the reference intervals are respectively established according to the prac‐tical situation ,which are suitable for this laboratory by verification .

Objective To investigate the related factors of early adverse outcomes in young patients with ischemic stroke.Methods From January 2006 to June 2016,685 young patients (18-45 years old) with acute ischemic stroke admitted to the Department of Neurology,Nanjing General Hospital of Nanjing Military Command were enrolled retrospectively.They were diagnosed as the first onset with head CT or MRI.According to the modified Rankin scale (mRS) at 90 d,the patients were divided into a favorable outcome (mRS 0-2) group (n=554) and a poor outcome (mRS 3-6) group (n=131).The collection of clinical data were completed on the day of admission,including the risk factors for cerebrovascular disease (oral contraceptives,etc),the National Institutes of Health stroke scale (NIHSS) score on admission,the mean systolic blood pressure (>140 mmHg was analyzed) and laboratory examination.The stroke subtypes were classified with the trial of org 10172 in acute stroke treatment (TOAST) classification criteria.Univariate analysis was used to analyze the difference of clinical data between groups,and multivariate logistic regression analysis was used to analyze the risk factors for early poor outcomes.Results Compared with the favorable outcome group,the patients with the ratio of mean systolic pressure >140 mmHg in the first 3 d after hospitalization (37.4% [49/131] vs.21.7% [120/554],χ2=14.131),NIHSS score on admission (10.0 [7.0,14.0] vs.1.5 [0,3.0],Z=-15.300),white blood cell count (7.5 [6.0,9.0] ×109/L vs.6.8 [5.7,8.2] ×109/L,Z=-3.157),fasting glucose (4.9 [4.6,6.0] mmol/L vs.4.8 [4.4,5.3] mmol/L,Z=-2.726),higher fibrinogen level (2.8 [2.3,3.4] g/L vs.2.6 [2.3,3.2] g/L,Z=-2.018,blood uric acid level (291[220,346] mmol/L vs.315 [261,374] mmol/L,Z=-3.443),and plasma albumin level (43.1[40.0,45.9] g/L vs.44.8 [42.4,47.4] g/L,Z=-4.708) were decreased in the poor outcome group.There were significant differences between the two groups (all P<0.05).TOAST classification comparison:the proportion of the patients with cardioembolism in the poor outcome group was higher than that in the favorable outcome group.There was significant difference between the two groups (6.9% [9/131] vs.2.5% [14/554];χ2=4.893,P<0.05).There were no significant differences in the remaining clinical data between the two groups (all P>0.05).Multivariate logistic regression analysis showed that the higher NIHSS score on admission (OR,1.474,95%CI 1.378-1.576,P<0.01),the mean systolic pressure >140 mmHg at the first 3 d after admission (OR,2.134,95%CI 1.210-3.764,P=0.009) and the cardioembolism(OR,4.902,95%CI 1.073-22.222,P=0.040) were the risk factors for early poor outcome,and the elevated plasma albumin level (OR,0.902,95%CI 0.850-0.956,P=0.001) was a protective factor of early favorable outcome.Conclusion The higher NIHSS score at admission,the cardioembolism and the increased mean systolic blood pressure in the first 3 d after admission may result in early poor outcome in young patients with ischemic stroke,while the elevated plasma albumin level is beneficial to the early outcome.

OBJECTIVETo evaluate the risk factors associated with pathogenesis of cerebral palsy (CP) in young children.

METHODSA cross-sectional survey on the prevalence of CP was conducted in young children aged one to six years in Leshan area, Sichuan Province. Cluster sampling and a 1:2 case-control design were used to investigate the risk factors for pathogenesis of CP.

RESULTSTotally, 148,723 children were surveyed, in which 308 (2.07 per thousand ) were diagnosed as CP. Low birth weight, twins and premature birth were associated with significantly increased prevalence of CP, 16.32, 4.16 and 22.21 times as that in normal birth weight, single birth and full-term birth, respectively. Simple factor analysis showed varied factors involved in pathogenesis of CP. The multivariate analysis revealed that delivery at home, low Apgar score in five minutes, illness during the first month of life, maternal "cold" with fever in their early gestation, low protein (meat and egg) intake during pregnancy and lower education level of mother were risk factors for CP.

CONCLUSIONSPrevalence and clinical features of CP in Leshan was comparable to those in the developed countries. Relevant risk factors could be seen primarily in gestational and perinatal periods, which might involve in mothers, children, environment and heredity, etc. To attach more importance in gestational and perinatal care for mothers and babies will be crucial measures to reduce occurrence of CP in young children.

Case-Control Studies ; Cerebral Palsy ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Infant, Premature ; Multivariate Analysis ; Prevalence ; Risk Factors

Objective To detect the level of 5-hydroxymethyl-cytosine (5-hmc)in melanoma tissues,and to analyze the correlation between 5-hmc and the invasion,metastasis and prognosis of melanoma.Methods A streptavidin-peroxidase immunohistochemical method was used to detect the level of 5-hmc in 67 melanoma tissues and 20 pigmented nevi tissues.Univariate and multivariate analyses were performed with the Cox's proportional hazards regression model to analyze the correlation between the expression of 5-hmc and the prognosis of melanoma.Results The expression rate of 5-hmc was significantly lower in melanoma tissues than in pigmented nevus tissues (40.30％ [27/67] vs.75％ [15/20],22 =7.428,P =0.006).According to American Joint Committee on Cancer (AJCC) TNM staging system,the expression level of 5-hmc was significantly lower in the stage Ⅳ melanoma tissues than in the stage Ⅱ and stage Ⅲ melanoma tissues (x2 =4.416,P =0.036).Patients with lymph node metastasis showed significantly lower expression of 5-hmc compared with those without lymph node metastasis (x2 =5.902,P =0.015),and the level of 5-hmc expression significantly decreased along with the increase of Clark grade (x2 =4.828,P =0.028).There were no significant differences in the level of 5-hmc expression between patients of different ages,genders or nationalities (P ＞ 0.05).Multivariate Cox regression analysis showed that distant lymph node metastasis (HR:2.67,95％ CI:1.22-5.84),not receiving surgical resection (HR:0.41,95％ CI:0.18-0.95),and low expression of 5-hmc (HR:3.54,95％ CI:1.09-11.43)were independent risk factors for poor prognosis of melanoma.Conclusion 5-Hmc may participate in the invasion and metastasis of melanoma,and be associated with the prognosis of melanoma.