Objective:To explore the role of retinoic acid-related orphan receptor α (RORα) in cognitive impairment induced by chronic alcohol consumption in mice.Methods:(1)The SPF grade RORαflox/flox transgenic mice aged 8 weeks were generated, and 22 transgenic mice were evenly divided into two groups by the method of matching body mass, which were the control group (Con group) and the alcohol group (EtOH group), with 11 mice in each group.(2)Emx1-Cre transgenic mice were used to selectively knock out the RORα gene in the forebrain of RORαflox/flox transgenic mice, producing conditional knockout mice (cKO mice) with the genotype RORαflox/flox-Emx1-Cre+ /+. Fourteen cKO mice were further split into two groups by the method of matching body mass, which were the conditional knockout group (cKO group) and the conditional knockout + alcohol group (cKO + EtOH group), with 7 mice in each group. A chronic alcohol use cognitive impairment model was developed in the EtOH group and cKO + EtOH group through the two-bottle free-choice method, while the Con group and cKO group were given two bottles of water for the same period. Cognitive abilities of mice in all groups were evaluated using behavioral novel object recognition test and Y-maze test.RORα mRNA and protein expression levels in the hippocampus of the Con group and EtOH group were assessed by RT-qPCR and Western blot, respectively.The GraphPad Prism 9.0 software was used for data analysis.One-way ANOVA was used for the comparison of multiple groups and Tukey test was used for further pairwise comparisons.Results:(1) Comparison between Con group and EtOH group: the relative levels of ROR α protein (0.63±0.04) and mRNA (0.78±0.03) in the hippocampus of mice in the EtOH group were significantly lower than those in the Con group((1.00±0.06), (1.00±0.05), both P<0.05). The duration of the EtOH group in the Y maze was significantly lower than that of the Con group ((212.30±32.05) s, (129.30±21.50) s, P<0.05), and the new object recognition index of the EtOH group was lower than that of the Con group ((14.73±25.49)% vs (-15.55±27.88)%, P=0.08). (2)Comparison between Con group and cKO group: the frequency and duration of entering the Y maze of mice in the cKO group ((7.43±2.30) times, (124.10±13.95) s) were lower than those in the Con group ((14.90±3.65) times, (212.30±32.05) s, both P<0.05). There was no statistically significant difference in the new object recognition index between the cKO group and the Con group ( P>0.05). (3) Comparison between the cKO+ EtOH group and the cKO group: the frequency ((2.71±1.11)times) and duration ((161.70±17.95) s) of entering the new heteroarm of Y maze in the KO+ EtOH group were lower than those in the cKO group ((7.43±2.30) times, (124.10±13.95) s, both P<0.05), and there was no statistically significant difference in the new object recognition index ( P>0.05). (4) Comparison between the cKO+ EtOH group and the EtOH group: the frequency of entering new heteroarm of the Y maze in the cKO+ EtOH group ((2.71±1.11)times) was significantly lower than that in the EtOH group (12.18±4.49) ( P<0.05), while there was no statistically significant difference in other behavioral results between the two groups ( P>0.05). Conclusion:Chronic alcohol consumption leads to cognitive impairment through the downregulation of RORα in the hippocampus of mice. Specific knockout of RORα in the forebrain exacerbates cognitive impairment induced by chronic alcohol use. RORα may represent a potential therapeutic target for cognitive impairment resulting from chronic alcohol consumption.

Objective:To explore the role of retinoic acid-related orphan receptor α (RORα) in cognitive impairment induced by chronic alcohol consumption in mice.Methods:(1)The SPF grade RORαflox/flox transgenic mice aged 8 weeks were generated, and 22 transgenic mice were evenly divided into two groups by the method of matching body mass, which were the control group (Con group) and the alcohol group (EtOH group), with 11 mice in each group.(2)Emx1-Cre transgenic mice were used to selectively knock out the RORα gene in the forebrain of RORαflox/flox transgenic mice, producing conditional knockout mice (cKO mice) with the genotype RORαflox/flox-Emx1-Cre+ /+. Fourteen cKO mice were further split into two groups by the method of matching body mass, which were the conditional knockout group (cKO group) and the conditional knockout + alcohol group (cKO + EtOH group), with 7 mice in each group. A chronic alcohol use cognitive impairment model was developed in the EtOH group and cKO + EtOH group through the two-bottle free-choice method, while the Con group and cKO group were given two bottles of water for the same period. Cognitive abilities of mice in all groups were evaluated using behavioral novel object recognition test and Y-maze test.RORα mRNA and protein expression levels in the hippocampus of the Con group and EtOH group were assessed by RT-qPCR and Western blot, respectively.The GraphPad Prism 9.0 software was used for data analysis.One-way ANOVA was used for the comparison of multiple groups and Tukey test was used for further pairwise comparisons.Results:(1) Comparison between Con group and EtOH group: the relative levels of ROR α protein (0.63±0.04) and mRNA (0.78±0.03) in the hippocampus of mice in the EtOH group were significantly lower than those in the Con group((1.00±0.06), (1.00±0.05), both P<0.05). The duration of the EtOH group in the Y maze was significantly lower than that of the Con group ((212.30±32.05) s, (129.30±21.50) s, P<0.05), and the new object recognition index of the EtOH group was lower than that of the Con group ((14.73±25.49)% vs (-15.55±27.88)%, P=0.08). (2)Comparison between Con group and cKO group: the frequency and duration of entering the Y maze of mice in the cKO group ((7.43±2.30) times, (124.10±13.95) s) were lower than those in the Con group ((14.90±3.65) times, (212.30±32.05) s, both P<0.05). There was no statistically significant difference in the new object recognition index between the cKO group and the Con group ( P>0.05). (3) Comparison between the cKO+ EtOH group and the cKO group: the frequency ((2.71±1.11)times) and duration ((161.70±17.95) s) of entering the new heteroarm of Y maze in the KO+ EtOH group were lower than those in the cKO group ((7.43±2.30) times, (124.10±13.95) s, both P<0.05), and there was no statistically significant difference in the new object recognition index ( P>0.05). (4) Comparison between the cKO+ EtOH group and the EtOH group: the frequency of entering new heteroarm of the Y maze in the cKO+ EtOH group ((2.71±1.11)times) was significantly lower than that in the EtOH group (12.18±4.49) ( P<0.05), while there was no statistically significant difference in other behavioral results between the two groups ( P>0.05). Conclusion:Chronic alcohol consumption leads to cognitive impairment through the downregulation of RORα in the hippocampus of mice. Specific knockout of RORα in the forebrain exacerbates cognitive impairment induced by chronic alcohol use. RORα may represent a potential therapeutic target for cognitive impairment resulting from chronic alcohol consumption.

Objective To summarize the clinical treatment experience of carbapenem-resistant Klebsiella pneumoniae (CRKP) infection after renal transplantation in donation after cardiac death (DCD) era. Methods Clinical data of 10 donors and 17 recipients with CRKP infection after DCD renal transplantation from January 2015 to January 2019 were retrospectively analyzed. Both donors and recipients received bacterial culture and drug sensitivity test. Clinical manifestations, treatment and outcome of CRKP-infected recipients were recorded. Results Seven donors were infected with CRKP. After pretreatment, CRKP in 2 cases turned negative, CRKP in 5 donors did not turn negative. All renal grafts were treated with tigecycline+meropenem+voriconazole lavage to prevent infection. Among 17 recipients with CRKP infection, 11 cases were positive for blood culture, 10 positive for urine culture, 3 positive for sputum culture, 3 positive for incisional secretion and 3 positive for retroperitoneal drainage. Clinical manifestations included fever in 8 cases, rupture and hemorrhage of the transplant renal artery in 7 cases or thrombosis in the transplant renal artery in 1 case, bladder irritation sign in 3 cases and cough with brick red jelly-like sputum in 1 case, respectively. Five patients were treated with tigecycline+meropenem, 1 patient suffered from renal graft loss and 4 recipients died. Twelve patients were treated with ceftazidime-avibactam +meropenem, 3 patients presented with renal graft loss and 1 recipient died. Conclusions CRKP-infected donor is not the absolute contraindication of renal transplantation. Pretreatment of donor infection and early administration of sufficient sensitive antibiotics can cure CRKP infection and improve the clinical prognosis of renal transplant recipients.

Objective:To explore the risk factors of pulmonary infection in elderly (aged 60+ years) kidney transplant recipients.Methods:The clinical data were retrospectively analyzed for 119 elderly kidney transplant recipients from January 2010 to January 2019 . According to whether or not pulmonary infection occurred after renal transplantation, the recipients were divided into infected group (n=40) and non-infected group (n=79). Clinical data was analyzed for two groups. The relevant risk factors of gender, age, donor type, body mass index, history of smoking, preoperative dialytic time, preoperative dialysis, immune induction, immune maintenance, presence or absence of delayed graft function, leucopenia, serum creatinine before infection, venous hormone shock therapy or not, diabetic history before or after surgery, history of coronary heart disease, history of hepatitis B virus, prophylactic dosing of compound sulfamethoxazole, prophylactic valganciclovir or ganciclovir, were examined by univariate analysis and multivariate logistic regression analysis.Results:The incidence of pulmonary infection in elderly kidney transplant recipients was 33.6% (40/119). In infected group, 15 patients died of severe pulmonary infection with a mortality rate of 37.5%(15/40). History of smoking (OR=10.58, 95%CI: 1.98-56.40, P=0.006), venous hormone shock therapy (OR=25.06, 95%CI: 4.25-147.71, P<0.001) and preoperative dialytic time (OR=1.032, 95%CI: 1.003-1.062, P=0.033) were the risk factors for pulmonary infection in elderly kidney transplant recipients. Conclusions:The incidence and mortality of lung infection are higher in elderly kidney transplant recipients. Smoking history, venous hormone shock therapy and long preoperative dialytic are associated with pulmonary infection in elderly kidney transplant recipients.

Objective To investigate the clinical characteristics of DCD donor-derived CRKP infection and bleeding in kidney transplantation,and to summarize the experience of diagnosis,treatment and prevention.Methods A retrospective analysis was carried out from July 2016 to December 2017 in hospital,containing clinical data of 4 cases of CRKP-infected DCD donors and 7 cases of kidney transplantation recipients.Results In the CRKP culture of 4 cases of DCD donors,1 case was positive for blood culture,1 case was positive for urine culture,1 case was positive for sputum culture,and 1 case was negative for blood,urine and sputum culture.The corresponding 7 recipients were all positive for blood culture after renal transplantation,4 cases were positive for urine culture,3 cases were positive for sputum culture,and 5 cases were positive for perirenal drainage.Of the 7 patients,4 cases had renal artery hemorrhage,1 of them was died.The average bleeding time was 17.75 days after operation (14-19 days).In 7 patients with renal transplantation,CRP increasd.And in 3 cases of deaths,CRP was stably higher than normal.Meanwhile,CRP in 4 surviving patients gradually decreased to the normal range after effective anti-infection treatment.All 7 patients were treated with carbapenems;2 patients were dead without avibactam therapy;and 5 cases were treated with avibactam and carbapenems and survived,1 case died and 1 case had good renal function recovery.Conclusion Positive CRKP in blood,urine and sputum of DCD donors can lead to CRKP infection in kidney transplant recipients.Even if the body fluids of donors are all negative,the false negative results could not be excluded.Persistent or increased high-level CRP after operation is an early warning on CRKP infection.And CRP can be used as an indicator for evaluating the effectiveness of anti CRKP therapy.The combination of avibactam and carbapenem antibiotics is an effective regimen in the treatment of DCD donor-derived CRKP.

Although oxymatrine (OMT) has been shown to directly inhibit the replication of hepatitis B virus (HBV), limited research has been done with this drug. In the present study, the antiviral effect of OMT was investigated in an immunocompetent mouse model of chronic HBV infection. The infection was achieved by tail vein injection of a large volume of DNA solution. OMT (2.2, 6.7 and 20 mg/kg) was administered by daily intraperitoneal injection for 6 weeks. The efficacy of OMT was evaluated by the levels of HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B core antigen (HBcAg). The immunoregulatory activity of OMT was evaluated by serum ELISA and flow cytometry. Results shows that OMT at 20 mg/kg inhibited HBV replication, and it was more efficient than entecavir (ETV) in the elimination of serum HBsAg and intrahepatic HBcAg. In addition, OMT accelerated the production of interferon-(IFN-) in a dose-dependent manner in CD4T cells. Our findings demonstrate the beneficial effects of OMT on the enhancement of immunological function and in the control of HBV antigens. The findings suggest this drug to be a good antiviral therapeutic candidate for the treatment of HBV infection.

Aim To explore the protective effects and underlying mechanisms of Liu weiwuling Tablets (LW-WL)in concanavalin A (ConA)induced acute immu-nological liver injury in mice.Methods Mice were randomly divided into control,model,Bicyclol,LW-WL low dose (8 g·kg-1 )and LWWL high dose (16 g ·kg-1 )group.The medicattion was performed once daily for seven consecutive days,then the model of im-munological liver injury was prepared by intravenous injection of ConA (15mg·kg-1)in the tail of mice in each group except for the control group one hour after the last treatment.The pathological changes of liver tissues of mice were evaluated by HE staining with, and the levels of alanine amino transferase (ALT),as-partate aminotransferase (AST),and total bilirubin (TBIL)in serum were analyzed by colorimetric meth-od;the level of interleukin 12 (IL-12 ),interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleu-kin 4 (IL-4)and interleukin 10 (IL-10)in liver was measured by real-time quantitative polymerase chain reaction (RT-qPCR);the changes of Th1 (IFN-γ) and Th2 (IL-4)cells were observed by flow cytometric (FCM)analysis;the expression of Th1/Th2 transcrip-tion factor T-bet/GATA-3 in liver tissue was detected by Western blot.Results Compared with normal con-trol group,the serum ALT,AST and TBIL were signif-icantly increased in model group, the pathological damage of the liver tissue was severe,and the necrosis and apoptosis of hepatic cells were large, which showed that the model was successful .Compared with model group,both low and high dose of LWWL could significantly reduce ALT,AST,TBIL levels in serum induced by ConA;Th1 cells in the spleen decreased, while Th2 cells increased;the expressions of IL-12, IFN-γand TNF-αmRNA were significantly inhibited with IL-4 and IL-10 mRNA expression elevated in mouse liver tissue;the expression of GATA-3 protein was up-regulated,T-bet protein expression showing no significant changes.Conclusion LWWL could regu-late Th1/Th2 balance,thus inhibiting the acute immu-nity hepatic injury induced by ConA.

Objective To investigate the prevalence and distribution characteristics of Cyclospora cayetanensis and Cryptosporidium ssp. infection in diarrhea cases of Yunnan Province. Methods To collect fresh faeces from diarrhea cases in 7 counties/cities, examine the specimens by direct smear with iodine staining and modified acid-fast staining. Results The infection rate of C. cayetanensis and Cryptosporidium ssp. was 3.97% and 5.29%, respectively. The infection rate of the two pathogenic coccidians was as high as 10.64% and 8.51% in preschool children. C. cayetanensis was found in 3 counties and Cryptosporidium in 6 counties. Conclusion Both C. cayetanensis and Cryptosporidium ssp. are prevalent in Yunnan Province with the latter distributed more widely, and the two pathogens are more prevalent in preschool children.