Objective To investigate the feasibility of RNA interference(RNAi)strategy in mouse(embryonic stem)ES cells and study the possibility of RNAi in dermatological research.Methods Specific RNAi was introduced with EGFP gene as a target,either by in situ production of dsRNA from transient trans-fection of a plasmid harboring a547bp inverted repeat,or by direct transfection of dsRNA made by in vitro transcription.Gene silence of EGFP in EGFP-transgenic mouse was investigated by topical delivery of dsRNA.Results This longer dsRNA was capable of inducing a sequence-specific RNAi for the target gene in epi-some and in chromosome of ES cells.It was also shown that this sequence-specific RNAi was observed in EGFP-transgenic mouse by topical delivery of dsRNA.Conclusion These results suggest that ES cell possess RNAi activity by longer dsRNA.We first report the study on RNAi activity by topical delivery of specific dsRNA.Above findings offer a possibility to use dsRNAi for inhibition of gene expression in dermatological re-search.

The clinical data of 459 patients,who were first diagnosed as HIV/HBV co-infection from January 2007 to December 2013,were retrospectively analyzed.Among all patients,there were 89 cases with CD4 ＜ 50/μl,134 cases with CD4 50-200/μl and 236 cases with CD4 ＞ 200/μl,when HIV infection was diagnosed.In these three groups with different CD4 levels,the HBV DNA positive rates were 49.3％ (37/75),50.5％ (54/107) and 33.7％ (66/196);the HBV viral load were (6.37 ± 1.71) log10 copies/ml,(5.82 ± 1.86) log10 copies/ml and (4.36 ± 1.64) log10 copies/ml;the rates of abnormal liver function were 29.2％ (26/89),29.1％ (39/134) and 10.6％ (25/236);the occurrence rates of end-stage-liver-diseases were 16.9％ (15/89),14.9％ (20/134) and 5.1％ (12/236);the mortality rates were 10.1％ (9/89),9.7％ (13/134) and 3.8％ (9/236),respectively.The HBV DNA positive rates,HBV viral load,the rates of abnormal liver function,the occurrence rates of end-stage-liver-diseases and the mortality rates in CD4 ＞ 200/μl group were lower than that in CD4 ＜ 50/μl group and 50-200/μl group.The results suggest that for HIV and HBV co-infection patients,HBV replication level and prognosis of liver diseases are associated with CD4 + T lymphocyte count.

Objective To investigate the influence of hepatitis B virus (HBV) infection on efficacy of combined antiretroviral therapy (cART) in patients with acquired immunodeficiency syndrome (AIDS).Methods Seventy-eight subjects with human immunodeficiency virus (HIV)/HBV co-infection and 156 subjects with HIV mono-infection were included.CD4+ T cell count,HIV viral load,HBV-markers and liver functions were routinely tested.The differences in survival rate,as well as immunological and virological responses between the two groups (HIV/HBV co-infection group and HIV mono-infection group) during cART were compared.Categorical data were compared by Chisquare test,measurement data were compared by t test,and measurement data with abnormal distribution were compared by Mann-Whitney test.Results At month 42 of cART,HIV RNA levels and CD4+ T cell counts of the two groups were comparable.However,at month 48,54 and 60 of cART,the immunological and virological responses of HIV/HBV co-infection group were less favorable than those of HIV mono-infection group.At each time point of month 12,24,36,48 and 60 of cART,3 out of 13 subjects with HIV/HBV co-infection maintained hepatitis B e antigen (HBeAg)loss; the HBeAg seroconversion rates were 32.1％ (9/28),50.0％ (14/28),53.6％ (15/28),64.3％ (18/28) and 71.4％ (20/28),respectively (x2 =10.189,P=0.037) ; HBV DNA negative rates were 95.1％ (39/41),82.9％ (34/41),68.3％ (28/41),43.9％ (18/41) and 43.9％ (18/41),respectively (x2 =29.982,P=0.000); liver dysfunction rate was 32.1 ％ (25/78),51.4％ (38/74),33.8％ (22/65),47.9％ (23/48) and 6.7％ (3/45),respectively (x2 =28.053,P=0.000).Mortalities in HIV/HBV co-infected and HIV mono-infected individuals were 24.4％ (19/78) and 5.1 ％ (8/156),respectively (x2 =18.841,P＜0.01).Sixteen out of the 19 deaths (84.2 ％) in HIV/ HBV co-infected subjects died of end stage liver diseases.Conclusions HBV co-infection diminishes the long term efficacy of cART.End stage liver diseases are the primary cause of death in HIV/HBV co-infected subjects during cART.

Objective To explore the safety and effectiveness of diagnostic ultrasound associated with microbubbles to open the blood brain barrier(BBB).Methods Microbubbles were injected through caudal vein,the rat head was radiated by GE Vivid 7 diagnostic ultrasound immediately.The radiated depth was located in the basal ganglia assisted by magnetic resonance imaging(MRI)scanning.The degree of BBB opening was evaluated by enhanced MRI and Evans blue dyeing.The safety was inspected by observation of cell morphology under hematoxylin eosin(HE)staining.Results After the rat head radiated by diagnostic ultrasound with microbubbles,signal enhancement of the radiated area was observed on post contrast T1-weighted images.Red fluorescence of Evans blue was detected by fluorescence microscope in the same area.Normal cellular morphology and structural integrity were showed by HE staining.Conclusions The BBB of rat could be opened targetedly and noninvasively by diagnostic ultrasound associated with microbubbles.This may provide a new strategy for the drugs and stem cells treatment in the central nervous system diseases.

Objective To investigate the influence of safe sex education and antiretroviral therapy (ART) on human immunodeficiency virus (HIV) antibody sero-conversion status among HIV-discordant spouses.Methods Totally 1 258 HIV/acquired immunodeficiency syndrome (AIDS) cases and their spouses were enrolled and the related information was collected during 2005 to 2007.The HIV negative spouses were tested for HIV antibody once every 6 months.The effects of safe sex education and ART on sero-conversion status of HIV-discordant spouses were analyzed.The date were analyzed by x2 test.Results Without any intervention,505 out of 1 258 HIV/AIDS spouses were positive for HIV test,with the HIV natural spousal transmission rate of 40.1％.Among 442 blood source and 816 sexual source index HIV cases,HIV was positive in 103 and 402 of their spouses,respectively,with the HIV transmission rates between couples of 23.3％ and 49.3 ％,respectively.HIV transmission rate in sexual source group was higher than that in blood source group (x2 =80.421,P＜0.01).Among 608 male and 650 female index HIV cases,HIV was positive in 333 and 172 of their spouses,respectively,with the HIV transmission rates between couples of 54.8％ and 26.5％,respectively.Rate of HIV transmission from men to women was higher than that from women to men (x2 =104.770,P＜0.01).Among 753 HIV discordant couples,only 5 spouses had HIV sero-conversion (0.7％) after comprehensive intervention measures during 5 years of follow-up period.Among 31 HIV/AIDS patients who were only accepted safe sex education,3 of their spouses were HIV positive (9.7％).The transmission rate was lower than that in patients without any intervention (40.1％) (x2 =11.760,P＜0.01).Among 722 HIV/AIDS patients received ART,2 of their spouses were HIV positive (0.3 ％),which was lower than the transmission rate (9.7％) in ART-na(i)ve group (x2=39.821,P＜0.01).Conclusion Safe sex education should be implemented throughout the health management of HIV/AIDS patients and their spouses.If possible,early ART should be carried out to reduce the risk of HIV transmission between married couples.

Objective:To investigate the epidemic trend and risk change of acquired immunodeficiency syndrome (AIDS) complicated with malignant tumors after combination antiretroviral therapy (cART).Methods:The types of malignant tumors in patients with AIDS at different stages of cART were analyzed among anti-human immunodeficiency virus (HIV)-positive population in Hubei Province screened in National AIDS/HIV prevention and control information system from 1st January, 2004 to 31st December, 2018. The standardized incidence ratios(SIR) of malignant tumors in AIDS patients was analyzed based on the incidence of malignant tumors in the general population in Hubei Province or China in 2013. The changes in risks for development of malignant tumors in AIDS patients at different cART stages from 2004 to 2013 and 2014 to 2018 were compared.Chi-square test was used for statistical analysis.Results:Three hundred and twenty-three out of 22 994 AIDS patients were diagnosed with malignant tumors. Non-Hodgkin lymphoma(NHL) and cervical cancer were most common types in acquired immunodeficiency syndrome-defining cancers (ADC), while liver cancers and lung cancers were the most common types in non-acquired immunodeficiency syndrome-defining cancers (NADC). The overall risk of malignancy in AIDS patients was similar to that in the general population (SIR=1.06, χ2=0.62, P=0.426). However, the risks of Kaposi sarcoma, NHL, Hodgkin lymphoma, cervical cancer, and head and face cancers (excepting nasopharyngeal cancer) in AIDS patients were significantly higher than those in the general population (SIR=834.09, 9.65, 13.33, 5.22 and 2.94, respectively, χ2=11 747.27, 625.54, 56.65, 184.21 and 13.66, respectively, all P<0.01). The risks of lung cancer, colorectal anal cancer, stomach cancer and breast cancer in AIDS patients were significantly lower than those in the general population (SIR=0.33, 0.36, 0.43 and 0.45, respectively, χ2=33.43, 12.84, 9.01 and 7.21, respectively, all P<0.05). The SIR of cervical cancer, liver cancer and colorectal anal cancer from 2014 to 2018 were 4.06, 0.43 and 0.10, respectively, which were significantly lower than those from 2004 to 2013 (7.42, 1.96 and 0.84, respectively). The differences were all statistically significant ( χ2=5.39, 19.52 and 10.86, respectively, all P<0.05). Conclusions:At present, there are no significant differences of the incidences of malignant tumors between AIDS patients and general population, but the tumor types are different. The most common malignant tumors in this region are NHL and cervical cancer, which should be noted that HIV screening among patients with such tumors is conducive to comprehensive treatment to improve the efficacy.

Objective:To investigate the relationship between positive rate and titer of hepatitis B surface antibody (anti-HBs) and CD4 + T lymphocyte count level in human immunodeficiency virus (HIV) infected patients after hepatitis B virus (HBV) exposure. Methods:A total of 4 893 HIV-infected patients were admitted to Zhongnan Hospital of Wuhan University from January 2010 to December 2018. The demographic data, HIV-related diagnosis, treatment information, CD4 + T lymphocyte count and serum markers of HBV infection of HIV infected patients were retrospectively analyzed. The patients were grouped according to the CD4 + T lymphocyte count and serum markers of HBV infection, and the differences of anti-HBs positive rate and HBV exposure rate in patients with different CD4 + T lymphocyte counts were compared.The differences of CD4 + T lymphocyte count in patients with different titer of anti-HBs were compared. Statistical analysis was performed using chi-square test, analysis of variance or t test. Results:Patients with HIV infection were divided into CD4 + T lymphocyte count<200/μL group (3 293 cases), 200-500/μL group (1 200 cases) and CD4 + T lymphocyte count>500/μL group (400 cases). The HBV exposure rates in the three groups were 78.0%(2 569/3 293), 77.0%(924/1 200) and 76.2%(305/400), respectively. The anti-HBs positive rates were 38.2%(1 258/3 293), 53.8%(645/1 200) and 62.5%(250/400), respectively. The anti-HBs titers were (120.00±36.45) IU/L, (148.00±26.40) IU/L and (212.00±92.08) IU/L, respectively. The exposure rates of HBV in the three groups were similar ( χ2=0.992, P=0.609), but the positive rates and titers of anti-HBs were significantly different ( χ2=146.779 and F=45.362, respectively, both P<0.01). When the patients were grouped by anti-HBs titer, 2 740 cases were divided into anti-HBs negative group (<10 IU/L), 1 220 cases in low anti-HBs group (10-99 IU/L), 693 cases in medium anti-HBs group (100-499 IU/L) and 240 cases in high anti-HBs group (≥500 IU/L). The CD4 + T lymphocyte count levels of the four groups were (150.00±8.42)/μL, (185.00±7.08)/μL, (243.00±12.07)/μL and (308.00±22.60)/μL, respectively. The overall CD4 + T lymphocyte count levels among the four groups were significantly different ( F=68.479, P<0.01). Among the 90 HIV infected patients who received anti-retroviral therapy (ART), the anti-HBs titer increased from (91.96±21.87) IU/L to (200.76±56.43) IU/L after treatment, and the anti-HBs level before and after treatment was significantly different ( t=-2.542, P=0.035). Among 208 patients with negative HBV markers, no patients had hepatitis B surface antigen switched to positive when monitored for an interval time of (26.2±5.3) months. Conclusions:The risk of HBV exposure in patients with HIV infection is not significantly related to the disease stage, but the positive rate and titer of anti-HBs are significantly positively correlated with CD4 + T lymphocyte count level. The monitoring of anti-HBs and the serum markers of HBV infection in the same individual is conducive to the in-depth understanding of the protective effect of anti-HBs and the scientific evaluation of the risk of infection after HBV exposure.

The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF—a disease that affects salivary gland (SG) function—is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.

BACKGROUND: The derivation of salivary gland (SG) progenitors from pluripotent stem cells (PSCs) presents significant potential for developmental biology and regenerative medicine. However, the existing protocols for inducing SG include limited factors, making it challenging to mimic the in vivo microenvironment of embryonic SGs. METHODS: We reported a cocktail factor approach to promote the differentiation of mouse embryonic stem cell (mESC)-derived oral epithelium (OE) into SG progenitors through a three-dimensional co-culture method. Upon confirming that the embryonic SG can promote the differentiation of mESC-derived OE, we performed RNA sequence analysis to identify factors involved in the differentiation of SG progenitors. RESULTS: Our findings highlight several efficient pathways related to SG development, with frequent appearances of four factors: IFN-c, TGF-b2, EGF, and IGF-1. The combined treatment using these cocktail factors increased the expression of key SG progenitor markers, including Sox9, Sox10, Krt5, and Krt14. However, absence of any one of these cocktail factors did not facilitate differentiation. Notably, aggregates treated with the cocktail factor formed SG epitheliallike structures and pre-bud-like structures on the surface. CONCLUSION In conclusion, this study offers a novel approach to developing a differentiation protocol that closely mimics the in vivo microenvironment of embryonic SGs. This provides a foundation for generating PSC-derived organoids with near-physiological cell behaviors and structures.

Objective: To understand current situation and relations of childhood abuse and psychological capital, providing scientific basis for adolescent mental health promotion. Methods: Stratified cluster sampling method was used to select 1 894 students from junior high schools and senior high schools in Harbin. Questionnaire survey was conducted by using Childhood Trauma QuestionnaireShort Form (CTQ-SF) and Positive Psychological Capital (PPQ). Results: The total rate of childhood abuse among adolescents in the region was 98.3%. In addition to emotional abuse, scores of other dimensions of childhood abuse were higher for boys than for girls(P<0.01). Childhood abuse in rural area was higher than those in urban area except physical abuse(P<0.05). High psychological capital was observed among participants with boys higher in selfefficacy and resilience than that of girls(P<0.05). Psychological capital in urban students was higher than rural students(P<0.05). Except for the negative correlation between sexual abuse and resilience, all other dimensions of childhood abuse were negatively correlated with four dimensions of psychological capital. Stepwise regression analysis showed that emotional abuse, emotional neglect and physical neglect was negatively correlated with all dimensions of psychological capital(P<0.01); Sexual abuse showed negative association with selfefficacy and optimism(P<0.01). Conclusion Childhood abuse is closely related to psychological capital among adolescents in Harbin, suggesting exposure to childhood abuse might confer detrimental effects on psychological capital development.