1.Tetrandrine enhances the host antiviral response through cGAMP-mediated cGAS-STING signaling pathway
Zhiwen WANG ; Yaming ZHANG ; Zhihui XIE
Chinese Journal of Microbiology and Immunology 2024;44(1):17-26
		                        		
		                        			
		                        			Objective:To investigate whether tetrandrine could be used as an agonist of cGAMP to enhance the activation of cGAS-STING signaling pathway and analyze the antiviral function of tetrandrine.Methods:THP1-Lucia-ISRE and RAW-Lucia-ISRE cells were incubated with different doses of tetrandrine in combination with cGAMP, respectively. IRF3 reporter activity was analyzed by luciferase reporter assay. Western blot was used to detect the activation of cGAS-STING signaling pathway. The expression of IFN-β, CXCL10 and CCL5 at mRNA level was quantified by real-time quantitative PCR. The expression of IFN-β at protein level was assessed by ELISA. HeLa cells stably expressing STING-GFP gene (HeLa-STNG-GFP cells) were constructed and stimulated with tetrandrine and cGAMP, then puncta-like structures were imaged by ZEISS LSM780. THP1-Lucia-ISRE cells were infected with herpes simplex virus type 1 (HSV-1) in the presence or absence of tetrandrine or cGAMP. The antiviral function of tetrandrine was analyzed by Western blot and fluorescence intensity assay.Results:Tetrandrine enhanced cGAMP-mediated IRF3 responses and activated cGAS-STING signaling pathway in combination with cGAMP. Tetrandrine combined with cGAMP triggered STING translocation and the formation of puncta-like structures in HeLa-STNG-GFP cells. The titer of HSV-1, the expression of HSV-glycoprotein D/UL30 and the fluorescence intensity of HSV-GFP were all decline after treating HSV-1-infected THP1-Lucia-ISRE cells with tetrandrine and cGAMP.Conclusions:Tetrandrine combined with cGAMP activates cGAS-STING signaling pathway, thus enhancing the host antiviral response.
		                        		
		                        		
		                        		
		                        	
2.Epidemiological characteristics of Mycoplasma pneumoniae infection and diagnostic value of IL-6 and PCT detection in 1 122 pneumonia patients in Nanjing,China
Anran XIE ; Zhihui DONG ; Hongpan XU ; Han SHEN
Chinese Journal of Clinical Laboratory Science 2024;42(8):624-627
		                        		
		                        			
		                        			Objective To analyze the characteristics and epidemic trends of Mycoplasma pneumoniae(MP)infection among the pa-tients in our hospital,and explore the diagnostic values of interleukin-6(IL-6)and procalcitonin(PCT)detection for MP infection.Methods A retrospective analysis was conducted on the test results of MP nucleic acid and clinical data,including age,gender,sam-pling date,and treatment category of the 1 122 pneumonia patients in our hospital from September 1,2022,to August 31,2023.The positive detection rate of MP nucleic acid was analyzed in different genders,ages,visit categories,and seasons.Receiver operating characteristic(ROC)curves were plotted to analyze the diagnostic value of IL-6 and PCT levels,and their ratio values in Mycoplasma pneumoniae pneumonia(MPP).Results Among the 1 122 pneumonia patients,the overall positive detection rate of MP was 24.96%.The positive detection rates of MP nucleic acid in male and female patients were 23.28%and 27.29%,respectively,with no statistical-ly significant difference(P>0.05).The median age of MP nucleic acid-negative patients was 37 years,while that of MP nucleic acid-positive patients was 7 years,showing a statistically significant difference(P<0.01).MP nucleic acid-positive patients were mainly concentrated in the age groups of 1 to 2 years(20.37%),3 to 5 years(25.60%),and 6 to 8 years(67.40%).The positive detection rates of MP nucleic acid among 913 inpatients and 209 outpatients were 22.45%and 35.89%,respectively,with statistically significant difference(P<0.01).The highest MP nucleic acid positive detection rate was observed in autumn(42.47%).MP could be co-infected with five other respiratory pathogens in which human rhinovirus(HRV)was the most common(17 cases).ROC curve analysis showed that the IL-6/PCT ratio had the best diagnostic performance for MP infection in the patients with all ages,with an area under the curve(AUCROC)of 0.746,superior to IL-6 and PCT alone.For MP-susceptible patients aged 6 to 8 years,IL-6 had the highest predictive value for MP infection,with an AUCROC of 0.767.Conclusions MPP tends to be epidemic in autumn,with children aged 6 to 8 years being the main susceptible population.The IL-6/PCT ratio is expected to become an auxiliary diagnostic and prognostic evaluation indi-cator for MPP.IL-6 levels have the highest predictive value for MP infection in the susceptible population.
		                        		
		                        		
		                        		
		                        	
3.Research Advances in the Association Between Alzheimer's Disease and Double-Stranded RNA-Dependent Protein Kinase
Yi GONG ; Xingyang XIAO ; Yousheng HU ; Yiwei XIE ; Zhihui WU
Acta Academiae Medicinae Sinicae 2024;46(3):425-434
		                        		
		                        			
		                        			Alzheimer's disease(AD)is a severe threat to human health and one of the three major causes of human death.Double-stranded RNA-dependent protein kinase(PKR)is an interferon-induced protein kinase involved in innate immunity.In the occurrence and development of AD,PKR is upregulated and continu-ously activated.On the one hand,the activation of PKR triggers an integrated stress response in brain cells.On the other hand,it indirectly upregulates the expression of 3-site amyloid precursor protein cleaving enzyme 1 and facilitates the accumulation of amyloid-β protein(Aβ),which could activate PKR activator to further activate PKR,thus forming a sustained accumulation cycle of Aβ.In addition,PKR can promote Tau phosphorylation,thereby reducing microtubule stability in nerve cells.Inflammation in brain tissue,neurotoxicity resulted from Aβaccumulation,and disruption of microtubule stability led to the progression of AD and the declines of memory and cognitive function.Therefore,PKR is a key molecule in the development and progression of AD.Effective PKR detection can aid in the diagnosis and prediction of AD progression and provide opportunities for clinical treat-ment.The inhibitors targeting PKR are expected to control the activity of PKR,thereby controlling the progression of AD.Therefore,PKR could be a target for the development of therapeutic drugs for AD.
		                        		
		                        		
		                        		
		                        	
4.Psychological experience of patients with breast cancer during rehabilitation: a Meta synthesis of qualitative studies
Hong CHEN ; Xuecheng JIANG ; Fa XIAO ; Wenxin XIE ; Yuanyuan LUO ; Min LI ; Lyuting YAN ; Zhihui YANG
Chinese Journal of Modern Nursing 2024;30(13):1730-1737
		                        		
		                        			
		                        			Objective:To systematically integrate qualitative studies on psychological experience of breast cancer patients in rehabilitation period, so as to provide basis for improving psychological care and rehabilitation efficacy.Methods:Qualitative research on psychological experience of breast cancer patients during rehabilitation was electronically retrieved in databases, including PubMed, Embase, Web of Science, Cochrane Library, China National Knowledge Infrastructure, Wanfang data, and China Biology Medical disc. The search period was from database establishment to September 2023. The quality evaluation of the literature was conducted using the 2020 version of the quality evaluation criteria for qualitative research of the Joanna Briggs Institute Evidence-Based Health Care Center. The results were integrated using the aggregative Meta integration method.Results:A total of 18 articles were included. Sixty-six research topics were extracted and categorized into 10 new categories, forming three integrated results, namely, psychological adjustment of patients with negative emotions and rehabilitation needs in the process of coping, family and social factors affecting patient rehabilitation experience, and patients perceiving that the healthcare system was not yet perfect.Conclusions:Medical and nursing staff should pay attention to the psychological problems of breast cancer patients in the rehabilitation period, understand their rehabilitation needs, provide targeted counseling and services, pay attention to the family and social support factors of breast cancer patients, help them improve their quality of life, and actively promote rehabilitation.
		                        		
		                        		
		                        		
		                        	
5.A Preliminary Study on the Domestication and Breeding Methods of Experimental Cats
Yongping HE ; Meixian XIE ; Zhihui PANG ; Qingxiu LI ; Huahong HE ; Namin ZHANG ; Wei LI
Laboratory Animal and Comparative Medicine 2023;43(1):67-72
		                        		
		                        			
		                        			ObjectiveFocused on the laboratory animal domestication and breeding of domestic cats, to explore the feeding management methods and breeding techniques of experimental cats. MethodsSeven Chinese garden cats from three litters were introduced from the rural suburbs of Guangzhou, and a breeding seed colony was established. The cats were domesticated in captivity, bred, closed breeding and transmission according to the feeding and management methods of laboratory animal. The population reproduction, the number of pregnancies per year, the litter season, the birth and weaning quality of the cats, and the survival rate of weaning were statistically collected. ResultsThe young breeding cats were able to adapt to the cage feeding management. In the transmission breeding and the expanded breeding colony, the number of female cats pregnant with one, two or three litters a year accounted for 63.2%, 26.3% and 10.5%, respectively. The proportions of litters born from the 1st to the 4th quarters were 20.7%, 20.7%, 27.6%, and 31.0%. A total of 29 pregnancies and 101 kittens were got from 19 female cats, with an average of (3.5±1.33) kittens per litter. The birth weights of female and male cats were (89.31±13.69) g and (93.47±15.12) g, respectively. Sixty-seven kittens survived from weaning. The average survival rate was 60.86%, and the weaning weights of female and male cats were (361.62±82.77) g and (376.0±91.71) g, respectively. ConclusionDomestic Chinese garden cats can adapt to laboratory animal feeding and breeding rules, and have strong fertility. They can normally pregnant and breeding throughout the year. The kittens grow to 5-6 months of age can meet the weight requirements for the examination of pharmaceutical hypotensive substances, and can be used as experimental cats for pharmaceutical examination with clear origin. 
		                        		
		                        		
		                        		
		                        	
6.Protection of Zuoguiwan Against Apoptosis of Follicles in Rats Injured by 60Co-γ Rays: Based on PI3K/Akt/mTOR Signaling Pathway
Fenqin ZHAO ; Mingxia AN ; Xiaonan DING ; Jieying LIU ; Yan ZHAO ; Zhihui XIE ; Shuping LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(18):12-19
		                        		
		                        			
		                        			ObjectiveTo investigate the protective effect of Zuoguiwan against 60Co-γ ray-induced premature aging of rats based on the phosphatidylinositol-3-kinases/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway. MethodSixty sexually mature female SD rats were irradiated with 60Co-γ rays (6.0 Gy, LD40) for 24 h at one time. Then they were randomized into model group, Bujiale group (0.18 g·kg-1·d-1), Bujiale (0.09 g·kg-1·d-1) + high-dose Zuoguiwan group (23.625 g·kg-1·d-1), high-dose Zuoguiwan group (23.625 g·kg-1·d-1), medium-dose Zuoguiwan group (9.45 g·kg-1·d-1), and low-dose Zuoguiwan group (4.725 g·kg-1·d-1). The administration (once a day) lasted 21 days. Serum indexes [follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2)] of rats were detected by enzyme-linked immunosorbent assay (ELISA), and morphological changes of ovarian tissues were observed based on hematoxylin and eosin (HE) staining. The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and the protein expression of phosphorylated (p)-PI3K, p-Akt, p-mTOR, B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in ovarian tissues by Western blot. ResultCompared with normal group, model group demonstrated increase in serum FSH (P<0.01), decrease in E2 (P<0.05), and reduction of follicles and luteum in early ovary (P<0.01). Moreover, the elevation of apoptosis rate of granulosa cells (P<0.01), down-regulation of p-PI3K, p-Akt, p-mTOR, and Bcl-2 in ovarian tissue, and increase in expression of Bax were also observed in the model group as compared with the normal group (P<0.01). In comparison with the model group, the administration groups showed rise of the number of early ovarian follicles, decrease in the apoptosis rate of granulosa cells, increase in the expression of p-PI3K, p-Akt, p-mTOR, and Bcl-2, and down-regulation of Bax, particularly the Bujiale + high-dose Zuoguiwan group(P<0.05,P<0.01). ConclusionZuoguiwan protects radiation-damaged ovary by activating the expression of PI3K/Akt/mTOR protein in ovarian tissue, increasing Bcl-2, and inhibiting the expression of Bax. 
		                        		
		                        		
		                        		
		                        	
7.Effects and Mechanism of Jidesheng Anti-venom Tablet on Local Wound Inflammation and Systemic Inflammatory Response of Snake Bite Patients
Jie HU ; Chaojin YANG ; Xingsong HE ; Zheng MING ; Zhihui XIE
China Pharmacy 2021;32(17):2102-2107
		                        		
		                        			
		                        			OBJECTIVE:To investigate the effects and mechanism of Jidesh eng anti-venom tablet on local wound inflammation and systemic inflammatory response of snake bite patients. METHODS :Totally 64 patients with snake bite admitted to our hospital during Jun. 2018-Jun. 2020 were randomly divided into control group and observation group ,with 32 cases in each group. Both groups received routine treatment ,such as debridement ,drainage,flushing,sealing,anti-venom,anti-infection,anti-fibrinolysis and anti-shock. Observation group was additionally given Jidesheng anti-venom tablet for internal and external use ,for consecutive 7 d. Related indexes of systemic inflammatory response ,local wound condition ,hospital stay ,laboratory indexes of important organs,coagulation function index ,wound inflammatory cell counts ,serum levels of inflammatory cytokines and chemokine ,the occurrence of ADR were compared between 2 groups. RESULTS :After treatment ,most of related indexes of systemic inflammatory response (RR,HR and WBC ),local wound condition (local pain disappearance time ,wound detumescence time ), hospital stay ,laboratory indexes of important organs (AST,ALT,Scr,BUN,CKB,CK-MB),coagulation function index (t-PA, PAI-1,TAT,SFMC),wound inflammatory cell (macrophages,neutrophils,lymphocytes)count,serum levels of inflammatory cytokines(TNF-α,IL-1,IL-6,hs-CRP,NF-κB)and chemokine (MCP-1,CXCL-8)in 2 groups were significantly better than before treatment (P<0.05);most indexes of observation group were significantly better than those of control group (P<0.05). No severe ADR was found in 2 groups. CONCLUSIONS :Jidesheng anti-venom tablet as auxiliary treatment can significantly reduce the local wound inflammation and systemic inflammatory response of snake bite patients ;the mechanism is probably related to reducing the levels of chemokine MCP- 1 and CXCL- 8 and inflammatory cytokines hs-CRP and NF-κB.
		                        		
		                        		
		                        		
		                        	
8.Efficacy and safety study of Chinese botulinum toxin A 100U in patients with overactive bladder: a prospective, multicenter, double-blind and randomized controlled trial
Limin LIAO ; Huiling CONG ; Zhihui XU ; Enhui LI ; Zhiliang WENG ; Haihong JIANG ; Ben LIU ; Xiao HUANG ; Shujie XIA ; Wei WEN ; Juan WU ; Guowei SHI ; Yang WANG ; Peijun LI ; Yang YU ; Zujun FANG ; Jie ZHENG ; Ye TIAN ; Haodong SHANG ; Hanzhong LI ; Zhongming HUANG ; Liqun ZHOU ; Yunxiang XIAO ; Yaoguang ZHANG ; Jianlong WANG ; Xiaodong ZHANG ; Peng ZHANG ; Dongwen WANG ; Xuhui ZHANG ; Keji XIE ; Bin WANG ; Lulin MA ; Xiaojun TIAN ; Lijun CHEN ; Jinkai DONG
Chinese Journal of Urology 2021;42(6):414-422
		                        		
		                        			
		                        			Objective:To assess the efficacy and safety of 100 units of botulinum toxin A (BTX-A) intradetrusor injection in patients with overactive bladder.Methods:From April 2016 to December 2018, 17 tertiary hospitals were selected to participate in this prospective, multicenter, randomized, double-blind, placebo-controlled study. Two phases of study were conducted: the primary phase and the extended phase. This study enrolled patients aged 18 to 75 years who had been inadequately managed by anticholinergic therapy (insufficient efficacy or intolerable side effects) and had spontaneous voiding with overactive bladder. Exclusion criteria included patients with severe cardiac, renal and hepatic disorders, patients with previous botulinum toxin treatment for 6 months or allergic to BTX-A, patients with urinary tract infections, patients with urinary stones, urinary tract tumors, diabetes mellitus, and bleeding tendency. Eligible patients were randomly assigned to BTX-A group and placebo control group in a ratio of 2∶1. Two groups of patients received 20 intradetrusor injections of BTX-A 100U or placebo at the depth of the submucosal muscle layer respectively under cystoscope, including 5 injections at the base of the bladder, 3 injections to the bladder triangle, 5 injections each to the left and right walls and 2 injections to the top, sparing the bladder neck. As a placebo control group, patients received same volume of placebo containing no BTX-A and only adjuvant freeze-dried preparations for injection with the same method. A combination of gelatin, sucrose, and dextran served as adjuvants. Average micturition times per 24 hours, urinary incontinence (UI) episodes per day, average micturition volume per day, OAB symptom score(OABSS), and quality of life (QOL) score were recorded at baseline and the 2nd, 6th and 12th week after treatment. The primary efficacy endpoint was the change from baseline in the average micturition times per 24 hours at the 6th week after treatment. The secondary efficacy endpoints included the change from baseline in the average micturition times per 24 hours at 2nd and 12th week, as well as the change from baseline in the OABSS, QOL score, average frequency of urgency and UI episodes per day, urgency score, average micturition volume per day at 2nd, 6th and 12th week after treatment. Patients were followed for 12 weeks to assess adverse events (AEs). After assessed at week 12, if the micturition times has decreased less than 50% compared to baseline and the patient is willing to receive retreatment, then patients could enter the extended trial phase. In that phase, patients in both groups were injected with 100 units BTX-A from 12th week onwards and then followed up the same indicators for 12 weeks.Results:216 patients were enrolled in this trial (144 cases in the BTX-A group and 72 cases in the placebo control group). Baseline characteristics such as age (47.75±14.20 in the BTX-A group and 46.39±15.55 in the control group), sex (25 male/117 female in the BTX-A group and 10/61 in the control group), and disease duration (0.51 years in the BTX-A group and 0.60 years in the control group) were balanced between the two groups( P>0.05). A marked reduction from baseline in average micturition times per 24 hours was observed in all treatment groups at the 6th week and the reduction of the two groups was statistically different ( P<0.001 and P=0.008 respectively). Compared with the baseline, the average micturition times per 24 hours at the 6th week decreased from baseline by 2.40(0.70, 4.60)times for the BTX-A group and 0.70(-1.00, 3.30) times for the placebo control group respectively, and the difference between the two groups was considered to be statistically significant ( P=0.003). The change rates of average micturition times per 24 hours from baseline at the 6th week of the two groups were (16±22)% and (8±25)% respectively, and the difference between the two groups was statistically significant ( P=0.014). Compared with the baseline, the average micturition times per 24 hours at 2nd and 12th week decreased by 2.00(0.00, 4.00)and 3.30(0.60, 5.03)for the BTX-A group, 1.00(-1.00, 3.00)and 1.70(-1.45, 3.85)for the placebo control group respectively. The difference between two groups was considered to be statistically significant ( P=0.038 and P=0.012); the changes of average urgency times per day for the BTX-A group and the control group at the 2nd, 6th and 12th week were 2.00(0.00, 4.30)and 2.40(0.30, 5.00), 3.00(0.30, 5.70)and 0.70(-1.30, 2.70), 0.70(-1.30, 3.00) and 1.35(-1.15, 3.50), respectively. There were significant differences between two groups at the 2nd, 6th and 12th week, ( P=0.010, P=0.003 and P=0.025, respectively). The OABSS of the BTX-A group and the control group at the 6th week decreased by 1.00(0.00, 4.00)and 0.50(-1.00, 2.00) compared with the baseline, and the difference between the two groups was statistically significant ( P=0.003). 47 cases of BTX-A group and 34 cases of placebo control group entered the extended trial phase, and 40 and 28 cases completed the extended trial phase, respectively. The average micturition volume per 24 hours changed by -16.60(-41.60, -0.60)ml and -6.40(-22.40, 13.30)ml, (-35.67±54.41)ml and(-1.76±48.69)ml, (-36.14±41.51)ml and (-9.28±44.59)ml, (-35.85±43.35)ml and(-10.41±40.29)ml for two groups at the 12th, 14th, 18th and 24th week, and the difference between two groups was statistically significant at each follow-up time ( P=0.01, 0.006, 0.012 and 0.016, respectively). There was no significant difference in other parameters( P>0.05). However, adverse reactions after intradetrusor injection included increased residual urine volume (27 in the BTX-A group and 3 in the control group), dysuria (21 in the BTX-A group and 6 in the control group), urinary infection (19 in the BTX-A group and 6 in the control group), bladder neck obstruction (3 in the BTX-A group and 0 in the control group), hematuria (3 in the BTX-A group and 1 in the control group), elevated alanine aminotransferase (3 in the BTX-A group and 0 in the control group), etc. During the follow-up period, there was no significant difference in the other adverse events between two groups except the increase of residual urine volume( P<0.05). In the primary trial phase, among the 27 cases with increased residual urine volume in BTA group, only 1 case (3.70%) with PVR more than 300 ml; the PVR of 3 patients in the placebo group was less than 100 ml. The increase of residual urine volume caused by the injection could be improved or disappeared with the passage of time. Conclusions:Intradetrusor injection of Chinese BTX-A improved the average micturition times per 24 hours, the average daily urgent micturition times, OABSS, and average micturition volume per time, and reduced the adverse effects in patients with overactive bladder.Chinese BTX-A at dose of 100U demonstrated durable efficacy and safety in the management of overactive bladder.
		                        		
		                        		
		                        		
		                        	
9.Effect of dendritic cells on immune function regulated by programmed cell death-1/programmed cell death-ligand 1 in sepsis
Zongpei WANG ; Zhihui XIE ; Yujie ZHAO ; Tingting BU ; Anyong YU ; Song WANG
Chinese Critical Care Medicine 2021;33(9):1032-1039
		                        		
		                        			
		                        			Objective:To study the effect of programmed cell death-1/programmed cell death-ligand 1 (PD-1/PD-L1) regulating dendritic cells (DC) on the immune status of sepsis, and analyze the effect of PD-1/PD-L1 on prognosis.Methods:Twenty-five patients with sepsis in the intensive care unit (ICU) of the Affiliated Hospital of Zunyi Medical University from October 2018 to September 2019 were collected and followed up for 28 days. According to the 28-day survival of patients, patients were divided into survival group and death group. Among them, 10 cases were in the survival group and 15 cases were in the death group. Simultaneously, 20 healthy subjects in our hospital during the same period served as the healthy control group. Peripheral blood of patients with sepsis was taken within 24 hours after diagnosis, and the healthy control group was taken at the time of enrollment. Flow cytometry was used to detect the proportion of CD4 +T and CD8 +T cells, the ratio of T cell subsets (CD4/CD8), the expression of PD-1 on CD4 +T and CD8 +T cells, and the expression of PD-L1 and CD86 in DC. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-10 (IL-10) and tumor necrosis factor-α(TNF-α) in serum. Spearman correlation analysis was used to analyze the correlation between CD11c +PD-L1 and CD4 +PD-1, CD8 +PD-1, TNF-α, DC, CD11c +CD86, T cell subpopulation ratio, CD4 +T cells, CD8 +T cells, and IL-10. Binary Logistic regression was used to analyze the risk factors affecting the death of patients with sepsis, and receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of independent risk factors on the prognosis of patients. Results:The scores of acute physiology and chronic health evaluationⅡ (APACHEⅡ) and sequential organ failure assessment (SOFA) in the death group were higher than that in the survival group (APACHEⅡ score: 27.0±7.3 vs. 17.0±3.9, SOFA score: 15.1±4.1 vs. 10.7±2.7, both P < 0.05). The ratio of T cell subsets in the survival group and the death group was less than 1, the death group was lower than that in the survival group (CD4/CD8: 0.54±0.15 vs. 0.79±0.09, P < 0.05), and the ratio of T cell subsets in the healthy control group was greater than 1. Compared with healthy control group, the levels of CD4 +T cells, CD8 +T cells, CD11c +DC, CD11c +CD86, IL-10 and TNF-α in survival group and death group were significantly decreased, the level of CD4 +PD-1, CD8 +PD-1, CD11c +PD-L1 were significantly increased, and the changes in the above indicators in the death group were significant compared with the survival group [CD4 +T cells: 0.14±0.07 vs. 0.22±0.08, CD8 +T cells: 0.24±0.07 vs. 0.28±0.10, CD11c +DC: 0.84±0.14 vs. 0.93±0.03, CD11c +CD86: (58.83±20.77)% vs. (78.24±9.39)%, IL-10 (ng/L): 34.22±13.98 vs. 18.49±5.55, TNF-α(ng/L): 95.30±29.33 vs. 67.00±20.16, CD4 +PD-1: (39.58±10.08)% vs. (27.03±6.35)%, CD8 +PD-1: (38.77±11.91)% vs. (29.15±8.37)%, CD11c +PD-L1: (21.13±11.54)% vs. (12.11± 8.34)%, all P < 0.05]. Spearman correlation analysis showed that CD11c +PD-L1 was positively correlated with CD4 +PD-1, CD8 +PD-1, and IL-10 ( r values were 0.748, 0.713, 0.898, all P < 0.05), while was negatively correlated with DC, CD11c +CD86, T cell subpopulation ratio, CD4 +T cells, CD8 +T cells, and TNF-α( r values were -0.587, -0.906, -0.840, -0.706, -0.513, -0.820, all P < 0.05). Multivariate binary Logistic regression analysis showed that CD4 +T PD-1 was an independent risk factor for the prognosis of sepsis patients [odds ratio ( OR) = 1.463, 95% confidence interval (95% CI) = 1.032-2.074, P = 0.033]. ROC curve analysis showed that CD4 +TPD-1 had certain predictive value for the prognosis of patients with sepsis [area under ROC curve (AUC) = 0.857, 95% CI was 0.709-1.000, P < 0.01). When the best predictive value was 34.48%, the sensitivity, specificity, and accuracy were 66.7%, 90.0%, and 85.7% respectively. Conclusions:Up-regulation of PD-1/PD-L1 in peripheral blood could inhibit the activation and proliferation of DC, affect the activation of T cells, and induce immunosuppressive state. PD-1/PD-L1 can reflect the immune status of patients with sepsis. The expression of PD-1 on CD4 +T cells may have important significance for the evaluation of prognosis.
		                        		
		                        		
		                        		
		                        	
10. Effect of lyciumbarbarum polysaccharide on sex hormone and ovarian estrogen receptor in rats exposed to n-hexane
Xiuxian NI ; Zipei HUANG ; Qianhuai XIE ; Zhijian CHEN ; Fei TANG ; Lu HUANG ; Ridong CAI ; Zhihui ZOU ; Rian YU
China Occupational Medicine 2020;47(03):291-297
		                        		
		                        			
		                        			 OBJECTIVE: To investigate the effect of n-hexane on the level of sex hormones and expression of estrogen receptor(ER) in rats and the protective effect of Lyciumbarbarum polysaccharide(LBP) on n-hexane-induced reproductive toxicity. METHODS: Based on factorial design model of 4×2, specific pathogen free adult female SD rats were divided into control group and low-, medium-and high-n-hexane exposure groups, and each group was divided into non-LBP intervention and LBP intervention sub-group. There were 8 subgroups with 6 rats in each group. On the first day, the rats in the 4 groups were given intraperitoneal injection of n-hexane at 0, 675, 1 350 and 2 700 mg/kg body weight, respectively. On day 2-4, the rats in the non-LBP intervention subgroup were given intragastric administration of 0.9% sodium chloride solution, and the rats in the LBP intervention subgroup were given intragastric administration of LBP at 50 mg/kg body weight once a day. On the fifth day, all animals were sacrificed, and the levels of follicle stimulating hormone(FSH), luteinizing hormone(LH), estradiol, progesterone were detected by enzyme linked immunosorbent assay. The mRNA expression of Erα, Erβ and G protein coupled estrogen receptor 1(Gper1) was detected by real time fluorescence polymerase chain reaction, and the expression of ERα, ERβ and GPER1 protein was detected by Western blotting. RESULTS: i) In the absence of LBP intervention(i.e. simple n-hexane exposure), there was no significant difference in the level of serum FSH, LH, estradiol and progesterone in the 4 groups(P>0.05). The relative expression of Erβ mRNA in ovary of low dose group decreased, while the relative expression of proteins of ERα and GPER1 increased(P<0.05) when compared with the control group. The relative expression of Erα mRNA and GPER1 protein in the ovary of medium-and high-dose groups increased(P<0.05), while the relative expression of Erβ, Gper1 mRNA and ERβ protein decreased(P<0.05). The relative expression of ERα protein in ovary of high-dose group increased(P<0.05). ii) At the same dose of n-hexane exposure, the relative expression of Erα mRNA in ovary of rats in low dose group increased(P<0.05), while the relative expression of ERβ and GPER1 protein decreased in LBP intervention group compared with the no LBP intervention group(P<0.05). The relative expression of ERα and GPER1 protein in ovary of medium dose group increased(P<0.05), while the relative expression of Gper1 mRNA and GPER1 protein in ovary of high dose group decreased in LBP intervention group compared with the no LBP intervention group(P<0.05). CONCLUSION: n-Hexane can up-regulate the expression of ERα and GPER1 in rat ovary, but has no significant effect on female endocrine system. LBP may play a protective role in female reproductive system by up-regulating the expression of ERα and GPER1. 
		                        		
		                        		
		                        		
		                        	
            
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