This study aims to investigate the epidemic variation of porcine circovirus type2(PCV2)and its distribution in swine farms in Henan Province from 2022 to 2023.In this study,1 206 pig blood samples from 18 urban scale swine farms,and 318 pigs and environmental samples from 2 PCV2-positive swine farms in Henan Province were detected by fluorescence quantitative PCR.ORF2 gene sequencing and genetic evolution analysis were performed on the positive samples.The results showed that the positive rate of PCV2 was 7.05％(85/1 206)in Henan Province,11.15％in western Henan Province and 3.03％in southern Henan Province.The positive rate of PCV2 in spring was 11.60％(67/576).A total of 31 ORF2 gene sequences were obtained,of which 23 were PCV2d(74.19％),6 were PCV2a(19.35％),one was PCV2b(3.23％)and one was PCV2c(3.23％).The results of amino acid sequence comparison showed that the mutation sites mainly concentrated on amino acids at positions 40-68 and 180-209.The total positive rate of pig farm sam-ples was 37.42％(119/318),and the positive rate of pig samples from different types of pig houses was 60.71％(34/56),45.83％(11/24),35.09％(20/57),and 33.33(3/9)in order from high to low.The positive rate of environmental samples from high to low was 56.36％(31/55)in feeding system,46.67％(7/15)in feces system,12.50(1/8)in decontamination area,11.11％(2/18)in living area,and no PCV2 was detected in ventilation system.The highest positive rate was 45.00％(9/20)for the tools in the house,7.14％(1/14)for the surface and sole of the clothes after leaving the house,and no PCV2 was detected on the surface and sole of the clothes before entering the house.In this study,the epidemiological investigation of PCV2 in Henan Province and its distribu-tion in swine farms were carried out,and the main circulating strains,seasons,regions and gene mutations of PCV2 in H enan Province were identified.The distribution and transmission routes of PCV2 in positive pig farms were preliminarily analyzed and summarized,which provided data basis and reference for effective prevention and control of PCV and the establishment of accurate bio-safety prevention and control measures.

Objective : To investigate the prevalence of negative emotions in hospitalized youth patients with type 2 diabetes(T2DM) and its correlation with coping strategies and psychological resilience. Methods : 141 youth T2DM patients who met the research standards were selected. Blood glucose related indicators, blood pressure, body mass index(BMI), diabetes chronic complications screening results and other data were collected. The basic information and disease related information questionnaire, self-rating depression scale(SDS), self-rating anxiety scale(SAS), diabetes distress scale(DDS), medical coping modes questionnaire(MCMQ) and Connor-Davidson resilience scale(CD-RISC) were completed. Results: Among 141 hospitalized youth T2DM patients, 37.6% were combined with depression, 32.6% were combined with anxiety, and 35.5% were combined with diabetic distress(DD). Univariate analysis showed that systolic blood pressure(P<0.01), educational level, and the form of hospitalization expenses(P<0.05) were significantly correlated with depression. Marital status(P<0.01), family residence, blood glucose monitoring methods, and the last fasting blood glucose(P<0.05) were significantly correlated with anxiety. BMI, whether it was first diagnosed or treated(P<0.01), gender, occupation, disease course, weekly blood glucose monitoring frequency, and the presence of chronic complications(P<0.05) were significantly correlated with DD. In multivariate analysis, systolic blood pressure(P<0.01), educational level, and the form of hospitalization expenses were significantly correlated with depression, marital status(P<0.05) was significantly correlated with anxiety; BMI and weekly blood glucose monitoring frequency(P<0.01) were significantly correlated with DD. SDS, SAS, total scores and dimensions of DDS were negatively correlated with the total score and dimensions of CD-RISC(rs=-0.182--0.467, P<0.05 or 0.01), and positively correlated with the yielding coping strategies(rs=0.177-0.271,P<0.05 or 0.01). SAS,total scores and dimensions of DDS were positively correlated with avoiding coping strategies(rs=0.237-0.419,P<0.05 or 0.01). The total and dimensions of CD-RISC were positively correlated with facing coping strategies(rs=0.215-0.349,P<0.05 or 0.01),and negatively correlated with yielding coping strategies(rs=-0.234--0.325,P<0.01). Conclusion More than 30% of hospitalized youth T2DM may experience negative emotions such as depression,anxiety,and DD. The occurrence of negative emotions in such patients may be related to disease management or socio-economic issues such as systolic blood pressure,educational level,hospitalization expenses,marital status,BMI,and frequency of blood glucose monitoring,as well as decreased psychological resilience and negative coping strategies.

Objective:To analyze the clinical characteristics of inconsistent symptoms and signs of dry eye in patients with Sj?gren syndrome (SS).Methods:A case-control study was performed.Thirty-eight patients (38 eyes) who visited the dry-eye outpatient department at Peking University People's Hospital were enrolled from January to October 2021.The patients were divided into a non-SS (NSS) group (25 cases, 25 eyes) and a SS group (13 cases, 13 eyes) according to without or with SS.The data of right eyes were analyzed.The patients' subjective symptoms were scored and their objective clinical parameters were evaluated.Evaluation of subjective symptoms included Ocular Surface Disease Index (OSDI), Standard Patient Evaluation of eye dryness (SPEED) Questionnaire and Dry Eye Questionnaire-5 (DEQ-5).Objective clinical parameters included Schirmer Ⅰ test (SⅠt), tear film breakup time (TBUT), SICCA ocular staining score (OSS), National Eye Institute (NEI) score, Marx line score, non-invasive tear film breakup time (NIBUT), non-invasive tear meniscus height (NITMH), meibomian gland loss area ratio, the number of corneal subepithelial nerves, total nerve length, mean nerve length, maximum nerve length, minimum nerve length, nerve curvature, and number of dendritic cells.Differences in subjective symptoms and clinical parameters were compared between the two groups, and correlation analysis between ocular symptom scores and objective clinical parameters was performed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University People's Hospital (No.RDY2020-03).Written informed consent was obtained from each subject.Results:The OSDI, SPEED and DEQ-5 scores of SS group were 10.9(2.1, 23.1), 4.0(2.0, 7.0) and 7.0(3.5, 9.5), respectively, which were significantly lower than 37.5(26.0, 64.9), 10.5(7.0, 13.0) and 13.0(6.8, 14.3) of NSS group ( Z=-2.70, -3.01, -2.14; all P<0.05).TBUT was longer in the SS group than in the NSS group, and the difference was statistically significant ( Z=-2.10, P=0.038).No significant difference was found in SⅠt, OSS, Marx line score, NIBUT, NITMH, meibomian gland loss area ratio, the number of corneal subepithelial nerves, total nerve length, mean nerve length, maximum nerve length, minimum nerve length, nerve curvature, and number of dendritic cells (all P>0.05).The OSS of nasal conjunctiva was significantly higher in the SS group than in the NSS group ( Z=-2.32, P=0.023).There were no correlations between the subjective symptoms and objective clinical parameters in NSS group (all P>0.05).The SPEED score was positively correlated with the Marx line score of upper eye lid ( rs=0.573, P=0.041) and the OSDI score was negatively correlated with the minimum nerve length ( rs=-0.606, P=0.037) in SS group. Conclusions:Patients with SS dry eye have more nasal conjunctival staining and fewer subjective symptoms than NSS dry eye patients with the same signs, which manifests as a separation of symptoms and signs.

Objective:To analyze the clinical characteristics of inconsistent symptoms and signs of dry eye in patients with Sj?gren syndrome (SS).Methods:A case-control study was performed.Thirty-eight patients (38 eyes) who visited the dry-eye outpatient department at Peking University People's Hospital were enrolled from January to October 2021.The patients were divided into a non-SS (NSS) group (25 cases, 25 eyes) and a SS group (13 cases, 13 eyes) according to without or with SS.The data of right eyes were analyzed.The patients' subjective symptoms were scored and their objective clinical parameters were evaluated.Evaluation of subjective symptoms included Ocular Surface Disease Index (OSDI), Standard Patient Evaluation of eye dryness (SPEED) Questionnaire and Dry Eye Questionnaire-5 (DEQ-5).Objective clinical parameters included Schirmer Ⅰ test (SⅠt), tear film breakup time (TBUT), SICCA ocular staining score (OSS), National Eye Institute (NEI) score, Marx line score, non-invasive tear film breakup time (NIBUT), non-invasive tear meniscus height (NITMH), meibomian gland loss area ratio, the number of corneal subepithelial nerves, total nerve length, mean nerve length, maximum nerve length, minimum nerve length, nerve curvature, and number of dendritic cells.Differences in subjective symptoms and clinical parameters were compared between the two groups, and correlation analysis between ocular symptom scores and objective clinical parameters was performed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University People's Hospital (No.RDY2020-03).Written informed consent was obtained from each subject.Results:The OSDI, SPEED and DEQ-5 scores of SS group were 10.9(2.1, 23.1), 4.0(2.0, 7.0) and 7.0(3.5, 9.5), respectively, which were significantly lower than 37.5(26.0, 64.9), 10.5(7.0, 13.0) and 13.0(6.8, 14.3) of NSS group ( Z=-2.70, -3.01, -2.14; all P<0.05).TBUT was longer in the SS group than in the NSS group, and the difference was statistically significant ( Z=-2.10, P=0.038).No significant difference was found in SⅠt, OSS, Marx line score, NIBUT, NITMH, meibomian gland loss area ratio, the number of corneal subepithelial nerves, total nerve length, mean nerve length, maximum nerve length, minimum nerve length, nerve curvature, and number of dendritic cells (all P>0.05).The OSS of nasal conjunctiva was significantly higher in the SS group than in the NSS group ( Z=-2.32, P=0.023).There were no correlations between the subjective symptoms and objective clinical parameters in NSS group (all P>0.05).The SPEED score was positively correlated with the Marx line score of upper eye lid ( rs=0.573, P=0.041) and the OSDI score was negatively correlated with the minimum nerve length ( rs=-0.606, P=0.037) in SS group. Conclusions:Patients with SS dry eye have more nasal conjunctival staining and fewer subjective symptoms than NSS dry eye patients with the same signs, which manifests as a separation of symptoms and signs.

This study aims to investigate the epidemic variation of porcine circovirus type2(PCV2)and its distribution in swine farms in Henan Province from 2022 to 2023.In this study,1 206 pig blood samples from 18 urban scale swine farms,and 318 pigs and environmental samples from 2 PCV2-positive swine farms in Henan Province were detected by fluorescence quantitative PCR.ORF2 gene sequencing and genetic evolution analysis were performed on the positive samples.The results showed that the positive rate of PCV2 was 7.05％(85/1 206)in Henan Province,11.15％in western Henan Province and 3.03％in southern Henan Province.The positive rate of PCV2 in spring was 11.60％(67/576).A total of 31 ORF2 gene sequences were obtained,of which 23 were PCV2d(74.19％),6 were PCV2a(19.35％),one was PCV2b(3.23％)and one was PCV2c(3.23％).The results of amino acid sequence comparison showed that the mutation sites mainly concentrated on amino acids at positions 40-68 and 180-209.The total positive rate of pig farm sam-ples was 37.42％(119/318),and the positive rate of pig samples from different types of pig houses was 60.71％(34/56),45.83％(11/24),35.09％(20/57),and 33.33(3/9)in order from high to low.The positive rate of environmental samples from high to low was 56.36％(31/55)in feeding system,46.67％(7/15)in feces system,12.50(1/8)in decontamination area,11.11％(2/18)in living area,and no PCV2 was detected in ventilation system.The highest positive rate was 45.00％(9/20)for the tools in the house,7.14％(1/14)for the surface and sole of the clothes after leaving the house,and no PCV2 was detected on the surface and sole of the clothes before entering the house.In this study,the epidemiological investigation of PCV2 in Henan Province and its distribu-tion in swine farms were carried out,and the main circulating strains,seasons,regions and gene mutations of PCV2 in H enan Province were identified.The distribution and transmission routes of PCV2 in positive pig farms were preliminarily analyzed and summarized,which provided data basis and reference for effective prevention and control of PCV and the establishment of accurate bio-safety prevention and control measures.

OBJECTIVE To investigate the effects of Forsythia suspensa ethanol extract on the proliferation， migration and invasion of lung cancer cells NCI-H226. METHODS As research objects， lung cancer cells NCI-H226 were divided into control group， F. suspensa ethanol extract low-， medium- and high-concentration groups （5， 10， 20 mg/mL）， activator group [10 mg/mL F. suspensa ethanol extract+0.5 μmol/L nuclear factor kappa B （NF-κB） signaling pathway activator PMA]， inhibitor group （10 mg/mL F. suspensa ethanol extract+10 μmol/L NF-κB signaling pathway inhibitor BAY 11-7082） and positive control group （20 μg/mL cisplatin）. Except for the control group of cells without intervention， all other groups of cells were cultured with corresponding drugs for 24 hours； the proliferation， migration and invasion of cells were all detected， and the proliferation rate， migration rate， and the number of invading cells were also calculated； protein expressions of NF-κB p65， NF-κB inhibitory protein α （IκBα）， phosphorylated NF-κB p65 （p-NF-κB p65） and phosphorylated IκBα （p-IκBα） were determined. RESULTS Compared with control group， the proliferation rate， migration rate， and the number of invading cells as well as the protein expressions of p- IκBα and p-NF-κB p65 were decreased significantly in F. suspensa ethanol extract groups and positive control group （P＜0.05）. Compared with F. suspensa ethanol extract medium-concentration group， the proliferation rate， migration rate， and the number of invading cells as well as above protein expressions were all decreased significantly in inhibitor group （P＜0.05）， while those of activator group were increased significantly （P＜0.05）. CONCLUSIONS F. suspensa ethanol extract can inhibit the proliferation， migration and invasion of lung cancer cells NCI-H226， and the mechanism of which may be related to the inhibition of NF-κB signaling pathway.

Objective To investigate the developmental feasibility of early human fetal testes (<3 months) using xenografting technique and to acquire an accessible donor derivation that is essential for studying human germ cell development. Methods Nine testes from 10-13 weeks aborted fetus were grafted under the back skin of 6 castrated nude mice. Grafts were collected at different time point according to the growth of the donor tissues and the health condition of the recipients. Morphological and histological analyses were performed for the observation of the development of grafted immature testicular tissues. Results The mass of grafts was increased from about 5-7mg to 84.1mg (the biggest). Six of 9 testes were to be in developing. Histological observations showed a significant expansion of seminiferous tubules from (44.26±3.14)μm to (77.69±7.47)μm. Cells dispersedly distributed in seminiferous cords at the time of grafting migrated towards the basal part of seminiferous epithelium. Some germ cells with spermatogonium-like characteristics located on the basement membrane. Sertoli cells were in stages from immature into matured with abundant cytoplasm which were orderly arranged around spermatogonia forming a niche-like structure. Conclusion Testes from early aborted human fetus grafted under the back skin of castrated nude mice showed further development and therefore could be used as an easier accessible donor tissues for the investigation of human spermatogenetic mechanism.

OBJECTIVETo investigate the development of xenografted primitive human germ cells by using fetal testicular tissues as donor tissues and an immunodeficient mouse as the recipient.

METHODSTesticular tissue fragments of a 26-week fetus were grafted under the back skin of a castrated immunodeficient mouse. Grafts were taken out after 135 days and processed for morphological and histological analyses.

RESULTSThe mass of grafts grew from about 1 mm in diameter and 5 mg in wet weight to about 3 mm and more than 20 mg 135 days after grafting. Histological observations showed a significant expansion of seminiferous tubules after grafting (80 +/- 25 microm in diameter) in comparison with seminiferous cords at the time of grafting (60 +/- 15 microm in diameter). The seminiferous cords developed into seminiferous tubules with the epithelial border and lumen. After 135 days of grafting, most of the dispersedly distributed primitive Sertoli cells and germ cells migrated to the basal part of seminiferous epithelium, located on the basement membrane and few of germ cells differentiated into spermatogonia.

CONCLUSIONHuman fetal testicular tissues could survive and continuously develop after being xenograft into castrated immunodeficient mice.

Animals ; Fetal Tissue Transplantation ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Spermatids ; growth & development ; Testis ; cytology ; transplantation ; Transplantation, Heterologous

Objective:To explore the effect of the microenvironment induced by damaged mouse hepatic cells on the conversion of human umbilical cord blood-derived cells into hepatocyte-like cells. Methods: A hepatic injury-like microenvironment was mimicked using carbon tetrachloride damaged mouse hepatic cells, where mononuclear cells (MNC) from human umbilical cord blood were cultured in a compartment separated by trans-well membrane. Histochemical staining, reversed transcription-polymerase chain reaction (RT-PCR) and gene sequencing were performed for the information on the conversion of human umbilical cord blood MNC. Results: A number of PAS positive stained cells in MNC co-cultured with damaged mouse hepatic cells were observed after 72 h. Cells expressing mature hepatocyte markers, human albumin (hALB) and human GATA-4 (hGATA-4) mRNA were detected by RT-PCR, which was further confirmed with sequencing. Relevant control groups, MNC co-cultured with normal mouse hepatic cells and MNC cultured alone remained negative. Conclusion: The culture system using damaged mouse hepatic cells as stimulator could be a potential in vitro system for the conversion of human umbilical cord blood-derived cells into hepatocyte-like cells.

Objective To investigate the fertilization ability of spermatozoa derived from neonatal mouse testes grafted into immunodeficient mice by intracytoplasmic sperm injection(ICSI).Methods Neonatal Kun-ming mouse testes containing primitive spermatogenic cells as the only germ cells were grafted under the back skin of castrated BALB/c-nu/nu mice.Grafts were taken out after 8 weeks.The initiation and restoration of spermatogenesis in grafts was documented by histological analyses.The ultrastructure of spermatogenic cells in grafts was observed using transmission electron microscopy.The activity of acrosomal enzyme of spermatozoa was analyzed with gelatin substrate test.The fertilization ability of spermatozoa was determined by using intracytoplasmic sperm injection.Results Histological analysis showed complete restoration of spermatogenesis in the grafts after 8 weeks.1?10~(3) spermatozoa with normal morphological appearance in 1mg suspended graft tissue were counted.Spermatogonia and spermatocytes with normal ultrastructure were observed.Gelatin substrate test positive spermatozoa were found implicating the activity of acrosomal enzyme of the spermatozoa.Cleavages were observed both in the ICSI group and the ICSI plus electric stimulation group.Conclusion Spermatozoa with normal morphology and fertilization ability can be developed by allografting neonatal mouse testes under the back skin of immunodeficient mice.