1.Isolation,identification and pathogenic characterization of Eimeria labbeana in Zhejiang province
Lianfeng WU ; Xueqiu CHEN ; Xiaocui HUANG ; Kaiyue SONG ; Kaiyin SHENG ; Yi YANG ; Guangxu MA ; Aifang DU
Chinese Journal of Veterinary Science 2025;45(6):1150-1156,1168
Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8%(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67%similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.
2.Human PCSK9D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice
ABIDAN·ABUDURUSULI ; Xiaocui CHEN ; Yuanfeng CUI ; TUOLUONAYI·MIJITI ; Lihui DENG ; Bangdang CHEN
Basic & Clinical Medicine 2025;45(5):637-643
Objective To investigate the effect of mutation human proprotein convertase subtilism/kexin type 9(hPCSK9D374Y)in PCSK9 gene on methionine choline deficiency diet(MCD)-induced nonalcoholic steato-hepatitis(NASH)in mice.Methods Sixteen C57BL/6J wild-type mice were selected and randomly divided into the hPCSK9D374Y group and the control GFP group.MCD was fed for 6 weeks,and then the serum level of hepatic triglyceride,alanine aminotransferase(ALT)and aspartate aminotransferase(AST)was examined.Oil Red O and Sirius Red staining microscopy were used to identify hepatic lipid infiltration and fibrosis severity.F4/80-positive cell infiltration was analyzed using immunohistochemistry.Lipid synthesis and inflammatory response-related proteins were detected by Western blot and related mRNA expression was analyzed by RT-qPCR.Results Hepatic hPCSK9 protein and mRNA were significantly up-regulated,LDLR protein expression was down-regulated,and ser-um level of ALT and AST was significantly elevated in the hPCSK9D374Y group of mice(P<0.05).The degree of he-patic steatosis and fibrosis increased and F4/80-positive cells were significantly increased(P<0.01).FASN and SCD1 proteins were significantly up-regulated and PPARα was down-regulated in the hPCSK9D374Y group;The ex-pression of TLR4 and p-P65 was elevated,whereas the expression of Iκ Bα was decreased(P<0.001).RT-qPCR re-sults showed a significant increase of mRNA coding inflammatory factors TNF-α,IL-1β,IL-6,and MCP-1,and a significant up-regulation of fibrosis-associated mRNAs(collagen Ⅰα and collagen Ⅲα)was found(P<0.001).Conclusions Functionally acquired mutation in the PCSK9 gene(hPCSK9D374Y)exacerbates MCD-induced hepatic steatosis,inflammatory response and fibrosis in mice.
3.Isolation,identification and pathogenic characterization of Eimeria labbeana in Zhejiang province
Lianfeng WU ; Xueqiu CHEN ; Xiaocui HUANG ; Kaiyue SONG ; Kaiyin SHENG ; Yi YANG ; Guangxu MA ; Aifang DU
Chinese Journal of Veterinary Science 2025;45(6):1150-1156,1168
Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8%(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67%similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.
4.Human PCSK9D374Y exacerbates methionine choline deficiency diet-induced nonalcoholic steatohepatitis in mice
ABIDAN·ABUDURUSULI ; Xiaocui CHEN ; Yuanfeng CUI ; TUOLUONAYI·MIJITI ; Lihui DENG ; Bangdang CHEN
Basic & Clinical Medicine 2025;45(5):637-643
Objective To investigate the effect of mutation human proprotein convertase subtilism/kexin type 9(hPCSK9D374Y)in PCSK9 gene on methionine choline deficiency diet(MCD)-induced nonalcoholic steato-hepatitis(NASH)in mice.Methods Sixteen C57BL/6J wild-type mice were selected and randomly divided into the hPCSK9D374Y group and the control GFP group.MCD was fed for 6 weeks,and then the serum level of hepatic triglyceride,alanine aminotransferase(ALT)and aspartate aminotransferase(AST)was examined.Oil Red O and Sirius Red staining microscopy were used to identify hepatic lipid infiltration and fibrosis severity.F4/80-positive cell infiltration was analyzed using immunohistochemistry.Lipid synthesis and inflammatory response-related proteins were detected by Western blot and related mRNA expression was analyzed by RT-qPCR.Results Hepatic hPCSK9 protein and mRNA were significantly up-regulated,LDLR protein expression was down-regulated,and ser-um level of ALT and AST was significantly elevated in the hPCSK9D374Y group of mice(P<0.05).The degree of he-patic steatosis and fibrosis increased and F4/80-positive cells were significantly increased(P<0.01).FASN and SCD1 proteins were significantly up-regulated and PPARα was down-regulated in the hPCSK9D374Y group;The ex-pression of TLR4 and p-P65 was elevated,whereas the expression of Iκ Bα was decreased(P<0.001).RT-qPCR re-sults showed a significant increase of mRNA coding inflammatory factors TNF-α,IL-1β,IL-6,and MCP-1,and a significant up-regulation of fibrosis-associated mRNAs(collagen Ⅰα and collagen Ⅲα)was found(P<0.001).Conclusions Functionally acquired mutation in the PCSK9 gene(hPCSK9D374Y)exacerbates MCD-induced hepatic steatosis,inflammatory response and fibrosis in mice.
5.Youguiwan Reduces Airway Inflammation in COPD Rats with Syndrome of Kidney-Yang Deficiency by Inhibiting Leptin/JAK2/STAT3 Signaling Pathway
Lan ZHENG ; Zeyuan LUO ; Min XIAO ; Xiaocui JIANG ; Yuhao MENG ; Siyi CHEN ; Jing ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):17-26
ObjectiveTo observe the effect of Youguiwan on the leptin/Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in the lung tissue of the rat model of chronic obstructive pulmonary disease (COPD) due to kidney-Yang deficiency. MethodForty rats were modeled for COPD with the syndrome of kidney-Yang deficiency by intratracheal instillation of lipopolysaccharide on day 1 and day 14 and continuous fumigation for 6 weeks, during which hydrocortisone was injected intramuscularly at an interval of 3 days. The modeled rats were randomized into model, high- (11.7 g·kg-1), medium- (5.85 g·kg-1), and low-dose (2.93 g·kg-1) Youguiwan, and aminophylline (0.054 g·kg-1) group. In addition, 8 SD rats were set as the blank group. After the completion of modeling, the rats in each group were administrated with the corresponding drug by gavage for 28 consecutive days. After the last administration, samples were collected. A lung function analyzer was used to evaluate the lung function of rats. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-17A (IL-17A), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the bronchoalveolar lavage fluid (BALF). Hematoxylin-eosin staining was employed to observe the pathological changes in the lung tissue, and Masson staining was employed to observe the deposition of blue collagen fibers around bronchi in the lung tissue and calculate the inflammation score. The immunofluorescence assay was employed to measure the protein content of collagen type Ⅰ (ColⅠ) and α-smooth muscle actin (α-SMA) in the bronchi. The protein and mRNA levels of leptin, IL-17A, JAK2, and STAT3 in the lung tissue were determined by Western blot and real-time fluorescence quantitative polymerase chain reaction, respectively. ResultCompared with the blank group, the model group showed decreased lung function (P<0.01), elevated levels of IL-6, IL-17A, and TNF-α in the BALF (P<0.01), and increased lung inflammation score, deposition of subcutaneous collagen fibers in the airway, and ColⅠ and α-SMA proteins (P<0.01). Furthermore, the modeling up-regulated the proteins and mRNA levels of leptin, IL-17A, JAK2, and STAT3 in the lung tissue (P<0.01) and enhanced the phosphorylation of JAK2 and STAT3 (P<0.01). Compared with the model group, high- and medium-dose Youguiwan improved the lung function, decreased the inflammation score, reduced collagen fiber deposition and ColⅠ and α-SMA proteins, lowered the levels of IL-6, IL-17A, and TNF-α in the BALF, down-regulated the mRNA and protein levels of leptin, JAK2, STAT3, and IL-17A, and weakened the phosphorylation of JAK2 and STAT3 (P<0.05, P<0.01). The aminophylline group had higher IL-17A and TNF-α levels than the high-dose Youguiwan group, lower IL-17A level than the medium and low-dose Youguiwan groups, and lower TNF-α level than the low-dose Youguiwan group. Compared with the aminophylline group, the high- and medium-dose Youguiwan groups showed reduced deposition of collagen fibers and protein levels of ColⅠ and α-SMA around the bronchi in the lung tissue (P<0.05, P<0.01), decreased inflammation score, and down-regulated protein and mRNA levels of leptin, JAK2, STAT3, and IL-17A in the lung tissue. ConclusionYouguiwan can prevent airway remodeling by inhibiting IL-17A to reduce inflammation and collagen deposition in COPD rats, which may be related to the inhibition of the leptin/JAK2/STAT3 signaling pathway.
6.Treatment of Asthenospermia and Infertility with Liver Constraint and Kidney Deficiency in 45 Cases by Shugan Bushen Yulin Decoction (疏肝补肾毓麟汤) and Vitamin E Soft Capsules: A Multicenter Randomized Controlled Trial
Wang WANG ; Jigang CAO ; Min XIAO ; Yangyang ZHU ; Mengjie GAO ; Qikai WANG ; Xiaocui JIANG ; Heng HAO ; Siyi CHEN
Journal of Traditional Chinese Medicine 2024;65(18):1909-1914
ObjectiveTo observe the clinical efficacy and safety of Shugan Bushen Yulin Decoction (疏肝补肾毓麟汤, SBYD) in the treatment of asthenospermia and infertility with liver constraint and kidney deficiency. MethodsA multicenter, randomized, controlled clinical study was conducted in three hospitals in central China. Totally 95 patients with asthenospermia and infertility were controlled. According to random number table, the patients were divided into treatment group (47 cases) and control group (48 cases). The control group was given vitamin E soft capsules (100 mg per time, twice daily) orally, and the treatment group was given SBYD (one dose daily, 30 min after breakfast and dinner, about 200 ml each time) orally. The course of treatment was 12 weeks in both groups. After the treatment, the sperm concentration, percentage of forward motile sperm (PR), and percentage of total sperm activity, that is PR + percentage of non-progressively motile sperm (NP) were compared between groups, and the clinical efficacy was judged. Traditional Chinese medicine (TCM) syndrome score (inlcuding 6 single symptom scores and total symptoms score) and 21-item depression, anxiety, and stress scale (DASS-21) scores (including depression, anxiety and stress scores) were compared between the two groups before and after treatment. The patients were followed up for 6 months, and the pregnancy status of spouse between groups was compared. The occurrence of adverse events and vital signs during the trial were recorded for safety assessment. ResultsTwo cases in the treatment group and three cases in the control group dropped out. Finally, 45 cases in each group were included in the statistical analysis. The total effective rate was 86.67% (39/45) in the treatment group, significantly higher than 73.74% (33/45) in the control group (P<0.05). After treatment, PR and PR + NP significantly increased in both groups, and were much higher in the treatment group than in the control group (P<0.05). The scores of lumbar and knee soreness, emotional disturbance, cold sperm, chest and rib-side and lesser abdomen distension and pain, frequent sighing, and the total TCM syndrome score in the treatment group decreased after treatment (P<0.05); and except for cold sperm, the scores of other symptoms mentioned above as well as the total TCM syndrome score in the treatment group were lower than those in the control group (P<0.05). The scores of depression, anxiety and stress of DASS-21 in the treatment group decreased after treatment, and were lower than those in the control group (P<0.05). After 6 months of follow-up, the spouses of 5 patients in the treatment group and 3 patients in the control group got pregnant (P>0.05). No adverse events occurred in both groups during the treatment, and their vital signs were stable. ConclusionSBYD can obviously improve sperm vitality and effectively improve the clinical symptoms, anxiety and depression, and is safe in treating asthenospermia and infertility with liver constraint and kidney deficiency.
7.A meta-analysis of related factors of depressive symptoms among men who have sex with men in China
Xiaocui ZOU ; Xiaorong MAO ; Xuemei CHEN ; Qinghua JIANG
Chinese Mental Health Journal 2024;38(2):186-192
Objective:To systematically review the related factors of depressive symptoms among men who have sex with men(MSM)in China.Methods:CNKI,Wanfang Data,VIP,CBM,EMbase,Pubmed,CINAHL,Web of Science and the Cochrane Library were searched from inception to February 17,2023.After literature screening and data extraction,two researchers independently assessed the quality of included studies according to the Agency for Healthcare Research and Quality.Results:A total of 21 articles with 11 822 participants were includ-ed.The results of meta-analysis showed that month income ≥3 000 yuan(OR=0.59),college degree or above(OR=0.59)and high self-esteem(OR=0.83)were protective factors,sexual role as the recipient(OR=1.68),dual sexual role(OR=1.41),multiple sexual partners(OR=1.65),sexual violence experience(OR=3.44),self-rated poor health status(OR=3.93),HIV/AIDS related discrimination(OR=1.13),HIV/AIDS related stress(OR=1.11)and suicidal tendency(OR=2.86)were risk factors for depressive symptoms.Conclusion:There are many related factors to the depressive symptoms of MSM.It is necessary to carry out early intervention on the basis of personalized assessment to reduce the occurrence of depressive symptoms among MSM.
8.Zuoguiwan Prevent Decreased Ovarian Reserve in Prenatal Stressed Offspring Rats via TLR4/NF-κB p65 Signaling Pathway
Yinjuan LYU ; Weiheng ZHANG ; Siyi CHEN ; Wenjing TIAN ; Jian GONG ; Yuhao MENG ; Huan HE ; Xiaocui JIANG ; Min ZHAO ; Min XIAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):67-76
ObjectiveTo observe the effect of Zuoguiwan on ovarian reserve in the female offspring rat model of prenatal stress (PS) and explore the mechanism based on Toll-like receptor 4/nuclear factor-κB p65 (TLR4/NF-κB p65) signaling pathway. MethodThirty-two pregnant rats were prepared and randomized into four groups (n=8): control, model, Zuoguiwan (18.9 mg·kg-1), and vitamin E (1.44 mg·kg-1). Except the control group, the other three groups were subjected to chronic unpredictable mild stress (CUMS) from day 11 of pregnancy, and the modeling was accompanied by gavage with corresponding drugs until delivery. The PS model was evaluated by the sucrose preference test, open field test, and serum corticosterone (CORT) level. The estrous cycle was monitored and the morphological changes in the ovarian tissue were observed. The serum levels of estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), and anti-Mullerian hormone (AMH) in the 75-day-old offspring rats were measured by enzyme-linked immunosorbent assay (ELISA) to evaluate the ovarian reserve. The ovary and uterus indices were calculated. The serum levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). The morphology of the ovarian tissue in the offspring on the day of birth and day 75 after birth was observed by hematoxylin-eosin staining. The transport of NF-κB p65 to the nucleus in the ovaries of the 75-day-old offspring was detected by the immunofluorescence (IF) assay. The expression of TLR4, NF-κB p65 and other related proteins in the ovarian tissue was determined by Western blot. ResultCompared with the control group, the model group showed reduced primordial follicles in the offspring on the day of birth (P<0.01) as well as disturbed estrous cycle, decreased ovary index and uterus index (P<0.01), reduced corpus luteum, increased atretic follicles (P<0.01), lowered serum levels of AMH and E2 (P<0.01), elevated serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and up-regulated protein levels of TLR4, NF-κB p65, recombinant myeloid differentiation factor 88 (MyD88), and phosphorylated NF-κB inhibitor (p-IκBα) (P<0.01) in the 75-day-old offspring rats. Compared with the model group, Zuoguiwan and vitamin E increased the primordial follicles in the offspring on the day of birth (P<0.01). Moreover, they resumed the estrous cycle, increased the ovary and uterine indices (P<0.05, P<0.01) and corpus luteum (P<0.01), reduced atretic follicles (P<0.01), elevated the serum levels of AMH and E2 (P<0.05, P<0.01), lowered the serum levels of LH, FSH, IL-1β, and TNF-α (P<0.05, P<0.01), and down-regulated the expression of TLR4, NF-κB p65, MyD88, and p-IκB-α (P<0.05, P<0.01) in the 75-day-old offspring. ConclusionZuoguiwan can improve the ovarian reserve in the offspring rat model of congenital kidney deficiency by regulating the TLR4/NF-κB p65 signaling pathway.
9.Establishment of a Predictive Model for Induced Abortion Failure in Term Sin-gle Pregnancy Pregnant Women
Ziyi GAO ; Zhenyu CHEN ; Xiaocui CHEN
Journal of Practical Obstetrics and Gynecology 2024;40(3):230-236
Objective:To analyze the influencing factors of failed induction of labor(IOL)in full-term singleton pregnant women,and to establish a prediction model for failed IOL.Methods:This study retrospectively analyzed the clinical data of 1483 pregnant women with full-term singleton of IOL in the Department of Obstetrics and Gy-necology,Heping Branch of General Hospital of Northern War Zone from January 1,2019 to December 31,2019.According to the outcome of IOL,the pregnant women were divided into the successful group(1108 cases)and the failed group(375 cases)of IOL.The influencing factors of failed IOL were screened to establish the prediction model through multivariate Logistic regression analysis.The receiver operating characteristic(ROC)curves and Hosmer-Lemeshow test were used to assess the predictive performance and fitting degree of the model.Results:Multivariate Logistic regression analysis showed that there were risk factors for failed IOL(OR>1,P<0.05),in-cluding elderly primiparous delivery,with no history of vaginal delivery,education level≤12 years,gestational age<40 weeks,pre-pregnancy overweight or obesity,excessive gestational weight gain,height<160 cm,cervical Bishop score before IOL<4 points,neonatal weight≥3750 g,combined IOL,suspected fetal distress,and the time from IOL to onset of labor≥24 hours,and height≥165 cm,IOL with dinoprostol suppositories were protective fac-tors for failed IOL(OR<1,P<0.05).Antepartum factors and antepartum factors combined with intrapartum fac-tors were separately used to establish model for predicting failed IOL.The area under the ROC curve(AUC)were 0.914 and 0.940,and the Youden index were 0.660 and 0.733,respectively.The prediction accuracy were 87.5%and 88.9%,respectively.Conclusions:This study screened the significant influencing factors of failed IOL,providing a theoretical basis for clinical measures to improve the success rate of IOL and constructing a pre-diction model of failed IOL,which is helpful for obstetricians and pregnant women to decide the mode of delivery together,and ensure the safety of mother and baby;on the other hand,it aims to enhance everyone′s awareness of pregnancy health care and improve the vaginal delivery rate.
10.Study on the contrast-enhanced spectral mammography and MRI in the dimension measurement of breast lesions
Meisha SUN ; Xiaocui RONG ; Jun CHEN ; Guang YANG ; Ning AI
Journal of Practical Radiology 2024;40(11):1801-1805
Objective To explore the accuracy and influencing factors of contrast-enhanced spectral mammography(CESM)and MRI in the dimension measurement of breast lesions.Methods A retrospective analysis was performed on 104 patients with breast lesions who underwent CESM and MRI examinations within one week.The pathological size was used as the gold standard to com-pare the accuracy of the two measurement methods.Logistic regression analysis was used to compare the influencing factors of the measurement accuracy of CESM and MRI.Results The measured values of CESM,MRI and pathology were 1.70 cm(1.20-2.70 cm),1.65 cm(1.20-2.28 cm),1.75 cm(1.00-2.50 cm),respectively,and there were no statistically significant differences between the two measurement methods and the pathological results(P>0.05).Bland-Altman plot analysis showed that 95.2%(99/104)of the differences in CESM fell within the consistency interval,and 93.3%(97/104)of the differences in MRI fell within the consistency interval.Logistic regression analysis showed that the independent risk factors affecting the accuracy of CESM and MRI measurement were lesion size and enhancement type.Conclusion Both CESM and MRI show good accuracy.As a potential alternative to MRI technology,CESM can be used for imaging evaluation of breast lesion size.

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