Anal Canal/surgery* ; Anorectal Malformations ; Constriction ; Fecal Incontinence ; Humans ; Rectal Prolapse

BACKGROUNDThe CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.

METHODSA SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.

RESULTSOne SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.

CONCLUSIONSWe conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.

Animals ; Chimera ; Genes, env ; HIV-1 ; genetics ; physiology ; Humans ; Macaca mulatta ; Proviruses ; genetics ; Receptors, CCR5 ; physiology ; Simian Immunodeficiency Virus ; genetics ; physiology

Objective To evaluate the outcome of health education program on drowning prevention among primary and secondary school children in rural areas. Methods A township was selected and all the students from grade 3 to 5, grade 7 to 8, and grade 10 to 11 were selected to take part in the program. Twelve intervention measures on natural water safety and drowning prevention were carried out for one year. Information was collected using the same questionnaire before and after the intervention program. Results One year after the intervention was carried out, children's knowledge on drowning prevention improved significantly (13.21, 95% CI: 12.51-13.90) , and a positive effect was also noticed among boys (12.77, 95%CI: 11.77-13.77), girls (13.80, 95%CI: 12.82-14.78),and among primary school children (15.51,95%CI: 14.30-16.72), senior high school children (10.78,95%CI: 9.50-12.05) and junior high school children (12.77,95%CI: 11.84-13.71). Overall rates on risk behaviors dropped from 41.4% to 32.2% (by 22.2%) including 15.6% for boys, 35.2% for girls and 13.8%, 29.3%, 26.3% for primary school children, senior high school children, junior high school children, respectively. The incidence rates for non-fatal drowning decreased by 58.9% (from 5.6% to 2.3%). The person-times for treatment on sight, in emergency settings, in outpatient clinic or in the hospitals had a reduction from 399, 78, 36 to 175, 32, 14, respectively. Conclusion Health education program could improve children's perception on water safety, and reduce their risk behaviors as well as on the incidence of non-fatal drowning in the rural areas.

Objective To analyze the relationship of cerebral microbleeds (CMBs)of different regions,especially mixed-CMBs,with cerebral amyloid angiopathy (CAA)detected using 18F-AV45 positron emission tomography(PET).Methods A total of 52 consecutive patients (68.17 ± 9.89 years old)with memory decline and CMBs found in susceptibility-weighted images(SWI)according to the inclusive and exclusive criteria were recruited.Patients were divided into three groups based on different regions of CMBs,the strictly lobar CMBs (SL-CMBs) group,the deep-CMBs (D-CMBs) group and the mixed-CMBs (M-CMBs)group.Patients in the three groups underwent 18F-AV45 PET detection and then were analyzed based on the results of 18F-AV45 PET.Results The positive rates of cerebral amyloid angiopathy in the SL-CMBs,M-CMBs and D-CMBs groups were 68.4 ％ (13/19),82.4 ％ (14/17) and 25.0 ％ (4/16),respectively,with statistical significance (P =0.002).There were significant differences in positive rates of cerebral amyloid angiopathy between the D-CMBs group and the M-CMBs group and between the D-CMBs group and the SL-CMBs group(P =0.001 and 0.010,respectively),while there was no difference between the M-CMBs and SL-CMBs groups in positive rates of cerebral amyloid angiopathy(P =0.335).Using the D-CMBs group as the reference group,multivariate logistic regression analysis showed that the odds ratios of positive CCA detected by PET in SL-CMBs and M-CMBs were 30.585(95％CI:2.492-375.360)and 8.107(95％CI:1.072-61.295),respectively.Conclusions Compared with D-CMBs,M-CMBs and SL-CMBs are more likely to be related to cerebral amyloid angiopathy.The presence of M-CMBs also indicates that patients have a high probability of CAA.

Objective:To analyze the characteristics of the apolipoprotein E(Apo E)genotype and cognitive impairment in patients with cerebral microbleeds(CMBs)and positive β-amyloid(Aβ)by using [18F]-AV45 positron emission tomography(PET).Methods:From September 2015 to May 2018, 152 patients with cognitive impairment and CMBs on the susceptibility-weighted imaging(SWI)sequence of head MRI at the neurology department of our hospital, assessed by mini-mental status examination(MMSE)score ≤26 and Montreal cognitive assessment(MoCA)≤25, were consecutively recruited in this retrospective study.After assessment with the inclusion and exclusion criteria, 69 patients aged 68.8±9.3 years were considered eligible for further analysis.Patients were divided into the Aβ-positive group(Aβ + Group, n=37)and the Aβ-negative group(Aβ -Group, n=32)after cognitive assessment, ApoE genotyping and [18F]-AV45 PET examination.Twenty-one healthy elderly controls(HC Group)who took health examination during the same period were enrolled.The results of cognitive assessment and Apo E genotyping were compared between the three groups. Results:The positive rate of the ApoE ε4 allele was 35.6%(32/90), 56.8%(21/37), 18.8%(6/32), and 23.9%(5/21)in the Aβ + , Aβ -and HC groups, respectively, with statistical significant differences between the groups( χ2=12.467, P<0.01). There were significant differences in the positive rate of the ApoE ε4 allele between the Aβ + and HC groups and between the Aβ + and Aβ -groups( χ2=5.880 and 10.407, P<0.05 and P<0.01). The percentage of patients with deep cerebral microbleeds was higher(56.3% or 18/32 vs.8.1% or 3/37, χ2=18.784, P<0.01)and of patients with lobar hemorrhage was lower(12.5% or 4/32 vs.45.9% or 17/37, χ2=9.066, P<0.01)in the Aβ -group than in the Aβ + group, while there was no significant difference in the percentage of patients with mixed cerebral microbleeds between the Aβ -and Aβ + groups( χ2=1.556, P>0.05). There were significant differences in cognitive function between the Aβ + and HC groups, in memory, executive function, visuospatial ability and language between the Aβ + and Aβ -groups, and in executive function, visuospatial ability and attention between the Aβ -and HC groups. Conclusions:Cognitive impairment is more extensive and severe in CMBs patients with Aβ deposition and is associated with positive ApoE ε4.

OBJECTIVETo investigate the effect of precursor of nerve growth factor (proNGF) in promoting invasion of breast cancer cells and its relation with ezrin expression and phosphorylation of ezrin Thr567 and Tyr477.

METHODSHuman breast cancer cell lines MDA-MB-231 and MCF-7 were stimulated by gradient concentrations of proNGF (0, 2.5, 5 and 10 ng/mL) for 16 h, and the invasion of the cells was assessed with Transwell assay. The expression of ezrin and the phosphorylation of ezrin Thr567 and ezrin Tyr477 in the treated cells were examined by Western blotting. MDA-MB-231 cells were transfected with pEnter-His-ezrinY477F (a dominant negative mutant) to study the role of phosphrylation of ezrin Tyr477 in the invasion of breast cancer cell stimulated by proNGF.

RESULTSproNGF significantly promoted MDA-MB-231 and MCF-7 cell invasion in a concentration-dependent manner (P<0.05), and concentration- and time-dependently increased the phosphorylation of ezrin Tyr477 (P<0.05) without affecting the expression of ezrin or the phosphorylation of ezrin Thr567. The specific inhibitor of src, SKI-606, significantly inhibited the phosphorylation of ezrin Tyr477 induced by proNGF. Transfection with pEnter-His- ezrinY477F inhibited proNGF-induced invasion and phosphorylation of ezrin Tyr477 in MDA-MB-231 cells (P<0.05).

CONCLUSIONPhosphorylation of ezrin Tyr477 plays a critical role in the invasion of breast cancer cells stimulated by proNGF via proNGF/src/ezrin Tyr477 pathway.

Breast Neoplasms ; pathology ; Cell Line, Tumor ; Cytoskeletal Proteins ; chemistry ; Humans ; MCF-7 Cells ; Neoplasm Invasiveness ; Nerve Growth Factor ; pharmacology ; Phosphorylation ; Signal Transduction ; Transfection ; Tyrosine

OBJECTIVETo study the relationship between Nanog-promoted metastasis of breast cancer and ezrin(T567) phosphorylation, and explore the possible mechanism by which Nanog regulates ezrin(T567) phosphorylation.

METHODSA siRNA construct targeting Nanog was transfected in breast cancer cells to knock down Nanog expression, and the changes in the cell invasion was detected using Transwell assay. The expression levels of Nanog and PKC and the phosphorylation level of ezrin(T567) were detected using Western blotting and immunofluorescent staining; the protein interaction between PKCε and ezrin was assayed by co-immunoprecipitation and Western blotting.

RESULTSNanog knockdown significantly decreased the expression of PKCε protein, phosphorylation level of ezrin(T567) and the invasion ability of breast cancer cells. PKCε knockdown obviously decreased the phosphorylation level of ezrin(T567) in the cells, and PKCε and ezrin were co-immunoprecipitated.

CONCLUDIONSNanogcan can upregulate the expression of PKCε to promote the phosphorylation of ezrin(T567), which can be a new mechanism by which Nanog promotes tumor metastasis.

Blotting, Western ; Breast Neoplasms ; metabolism ; Cytoskeletal Proteins ; metabolism ; Gene Knockdown Techniques ; Homeodomain Proteins ; metabolism ; Humans ; Nanog Homeobox Protein ; Neoplasm Invasiveness ; Phosphorylation ; Protein Kinase C-epsilon ; metabolism ; RNA, Small Interfering ; Transfection ; Tumor Cells, Cultured ; Up-Regulation

OBJECTIVETo study if programmed death-ligand 1 (PL-L1) expression in breast cancer cell activates PD-L1/PD-1 pathway in dendritic cells to inhibit dendritic cell maturation.

METHODSHuman monocytes were induced to differentiate into immature dendritic cells using GM-CSF and IL-4, and further to mature dendritic cells using TNF-α. PD-L1-expressing breast cancer cell line MDA-MB-231 was co-cultured in contact with the dendritic cells to observe the effects of the breast cancer cells on the maturation of the dendritic cells. A PD-L1 blocking antibody was applied to the co-culture, and the changes in the inhibitory effect of the MDA-MB-231 cells on dendritic cell maturation was observed. TNF-α-induced dendritic cells were treated with a recombinant human PD-L1 protein to study the effect of PD-L1/PD-1 pathway activation on the maturation of dendritic cells. The expression of PD-L1 in MDA-MB-231 cells and the dendritic cell maturation marker HLA-DR and CD83 were analyzed using flow cytometry.

RESULTSMDA-MB-231 cell line showed PD-L1 positivity on the cell membrane cells at a rate as high as (99.7∓0.15)%. In mature dendritic cells, the positivity rates for HLA-DR and CD83 were (88.8∓6.96)% and (18.36∓3.07)%, respectively, but in the co-culture system, the positivity rates of the dendritic cells were significantly decreased to (42.76∓10.52)% (P<0.01) and (9.93∓2.74)% (P<0.05), respectively, indicating that MDA-MB-231 cells inhibited the maturation of dendritic cells. Following treatment with a PD-L1 antibody isotype control, the percentages of HLA-DR- and CD83-positive cells in the co-culture were (45.17∓10.19)% and (10.15∓2.54)%, which were significantly increased to (63.46∓1.72)% and (16.46∓2.58)% after treatment with PD-L1 antibody, respectively (both P<0.05). Compared with the mature dendritic cell controls, the cells treated with the recombinant human PD-L1 protein exhibited significantly lowered percentages of HLA-DR-positive [from (84.23∓4.18)% to (2.56∓2.39)%, P<0.05] and CD83-positive cells [(87.26∓1.54)% to (60.67∓1.63)%, P<0.05].

CONCLUSIONThe effect of PD-L1 antibody therapy on triple negative breast cancer can be partially mediated by blocking PD-L1 expression on breast cancer cell membrane, which attenuates the inhibition of dendritic cell maturation in the cancer microenvironment.

OBJECTIVETo study the serum homocysteine (Hcy) distribution and characteristics in different sex and age groups in the community residents in Wuhan, and to analyse its associated factors with multi-stepwise regression analysis.

METHODSThe population under study was from three community areas in Wuhan. Demographic distribution and the correlation with other risk factors of serum Hcy were analyzed statistically.

RESULTS(1) Geometric mean of serum Hcy was 14.43 micromol/L in males and 10.89 micromol/L in females with P <0.001. (2) Hcy of per age level in males was also higher (P <0.001). (3) The prevalence rate of hyperhomocysteinemia was 23.94% in the general population in Wuhan. The prevalence rate of hyperhomocysteinemia in males was 2.62 times higher than in females. (4) Multi-stepwise regression analysis showed that Hcy had different affecting factors in males and females. The affecting factors of Hcy in males were daily cigarettes smoking, urine micro-albumin (UMALB) and times of exercise per week. The affecting factors of Hcy in females were duration of exercise each time, weight, triglyceride (TG), high-density lipoprotein (HDL), urine micro-albumin (UMALB) and age.

CONCLUSIONS(1) Hcy at the population level was significantly different by sex and age. (2) Population living in the community in Wuhan had a higher serum level and prevalence rate of Hcy comparing to some other cities in China and even in developed countries. (3) The important affecting factors of Hcy in population also showed sex difference, unlike the reports from other countries or other areas in China. Serum Hcy seemed to be affected by environmental and other factors.

Adolescent ; Adult ; Age Factors ; Aged ; China ; Female ; Homocysteine ; blood ; Humans ; Male ; Middle Aged ; Population Groups ; Reference Values ; Regression Analysis ; Sex Factors

OBJECTIVETo determine the association between viral load of high risk human papillomavirus (HR-HPV) and cervical intraepithelial neoplasia (CIN).

METHODSCervical exfoliated cells were collected from 18 186 women aged 17 -59 from six urban areas and eight rural areas when they were screened in the cross-sectional population-based studies from 1999 to 2008. HR-HPV was detected by the Hybrid Capture 2 (hc2) system, and viral load was measured by the ratio of relative light units to standard positive control (RLU/PC). RLU/PC was categorized for analysis into four groups: negative [0, 1.00), low viral load [1.0, 10.00), moderate viral load [10.00, 100.00), and high viral load > or = 100.00. Cervical lesions were diagnosed by biopsies as normal, CIN 1, CIN 2, CIN 3 and squamous cervical cancer (SCC). Association between HR-HPV viral load and CIN was evaluated by unconditional multinomial logistic regression.

RESULTSThe HR-HPV infection rate of the population was 14.51% (2515/17334). 100.00% (29/29) of SCC, 97.63% (206/211) of CIN 3, 93.43% (199/213) of CIN 2, 75.04% (421/561) of CIN 1 and 10.17% (1660/16320) of normal women were positive for HR-HPV DNA. The median RLUs for the HR-HPV positive women with SCC, CIN 3, CIN 2, CIN 1 and normal were 320.85, 158.05, 143.70, 125.34 and 9.64, respectively. There were significant differences among the distributions of viral loads in each lesion (chi2 = 6190.40, P < 0.01). The severity of CIN increased with the viral load (chi2 = 5493.35, P <0.01). Compared with the risks of CINs in HR-HPV negative population, the risks of CINs in low, moderate and high viral loads were increased gradually [OR(95% CI) : CIN 1 : 9.01(6.31 - 12.87), 24.96(18.23 - 34.17) and 68.42(51.40 - 91.08); CIN 2 : 26.44(12.07 - 57.95), 98.53(49.54 - 195.98) and 322.88(168.62 - 618.27); CIN 3+ : 72.89(24.02-221.18); 343.58(121.81-969.09) and >999.99(473.38 - >999.99)], and there were obvious dose-response relationships (chi2trend was 3115.05, 2413.95 and 3098.57, respectively. P< 0.01). In each age group of the HR-HPV positive population,the risks of CIN 2 + in the women with moderate or high viral load were higher than the one with low viral load [OR(95% CI): <35 : 4.71(1.23 - 18.09) and 15.06(4.40 - 51.49); 35 -: 4.01 (1.62 -9.90) and 14.09(6.15 -32.28); 40 - : 3.06(1.52 -6.16) and 7.78(4.05 -14.95); > or =45: 3.50(1.36 -9. 01) and 7.57(3.13 - 18. 30)], and there was a positive correlation between the risk of CIN 2+ and the viral load (chi2trend was 51.33, 66.28, 53.64 and 51.00, respectively. P <0.01). The risk of CIN 2 + was highest among the women aged 40 - with high viral load [OR (95% CI) : 2.02 (1.15 - 3.52)].

CONCLUSIONThere is strong correlation between the HR-HPV viral load and the severity of CIN, and so is the correlation between the HR-HPV viral load and the risk of CIN 2 +. A moderate to high viral load of HR-HPV should be the major risk factor for the cervical cancer and CIN 2 and CIN 3, and there is a higher risk in the women aged 35 or older than the younger ones. Considering both the age and viral load could help the doctors to manage the screening women more effectively.

Adult ; Cervical Intraepithelial Neoplasia ; epidemiology ; pathology ; virology ; Cervix Uteri ; pathology ; virology ; Female ; Humans ; Middle Aged ; Papillomavirus Infections ; epidemiology ; pathology ; virology ; Risk Factors ; Uterine Cervical Neoplasms ; epidemiology ; pathology ; virology ; Viral Load