To investigate the effects of ORF9 gene of porcine circovirus type 2(PCV2)on PK-15,eu-karyotic expression plasmid was constructed and transfected into PK-15 cells,and the effects of overexpression of ORF9 on proliferation,apoptosis and immunization of PK-15 cells were exam-ined by flow cytometry and qRT-PCR.The results showed that ORF9 gene overexpression signifi-cantly up-regulated the expression levels of the ER stress marker gene GRP78,increased the num-ber of S phase cells,accelerated cell cycle progression,increased the apoptosis rate of PK-15 cells,up-regulated the expression levels of apoptosis-related genes caspase-3,caspase-8,caspase-9,p53 and Bax(P＜0.01),down-regulated the expression levels of apoptosis-related genes Bcl-2,up-reg-ulated the expression levels of immune-related genes 1L-8,IL-10,NF-κB and TNF-α(P＜0.01),and down-regulated the expression levels of immune-related genes IL-2,IFN-β and IL-12(P＜0.01).The above results indicate that ORF9 gene may promote the proliferation and apoptosis of PK-15 cells and play a role in the escape process of PK-15 cells.

In order to understand the genomic characteristics and genetic variation and strain type of infectious bursal disease virus(IBDV)isolate GZGY2022,which caused the death of chickens in Guizhou farm,primers were designed to amplify the whole genome of the isolate,and genetic evo-lution and strain type analysis were performed after cloning and sequencing.The results showed that the A and B segments of IBDV genome were 3 260,2 827 bp,respectively,encoding VP2-VP5 and VP1 genes.The nucleotide sequence homology between the A and B segments of this strain and the VvIBDV were 96.2％-98.7％and 87.7％-98.9％,respectively,which is the highest with NN1172 strain,83.1％-94.7％and 90.1％-91.0％with other strains.The results of genetic evolution and strain type study showed that IBDV strains can be divided into 6 branches according to antigen and virulence,and the A and B segments of the strain were clustered in the evolutionary branch of VvIBDV,and the strain was A3B3 genotype according to the new genotype classification method.The results of amino acid sequence analysis showed that there were 3 and 7 unique amino acid site variations in the A and B segments of the strain,respectively,and 13 unique characteristic amino acid sites in the coding region of the full-length genome were consistent with VvIBDV.The VP2 sequence of segment A has 19 characteristic amino acid identical with VvIBDV,among which hyper variable regions 222A,242I,253Q,256I,279D,284A,294I and 299S were characteristic ami-no acid sites of the VvIBDV,and the heptapeptide region sequence SWSASGS was consistent with the virulent strain.The VP1 sequence of segment B has 10 characteristic amino acid identical with VvIBDV,among which 61I,145T and 287A were the characteristic amino acid sites of the VvIB-DV.In addition,the nucleotide sequence GGTGCC of 777-782 did not form the restriction endo-nuclease site of Kpn Ⅰ,and combined with the triplet site 145/146/147(TEG),the segment B was consistent with the NN1172 strain,showed that its virulence was slightly weaker than that of the B2 strain of VvIBDV.The results of recombination analysis showed that there were no breaks and recombination sites in the sequence of the strain,and no recombination event occurred.In summa-ry,this study found that GZGY2022 strain belonged to the A3B3 genotype non-recombinant VvIB-DV strain,and its special amino acid sites were consistent with the molecular characteristics of VvIBDV.This study lays the foundation for further exploring the genomic characteristics and path-ogenicity of VvIBDV.

Objective To investigate the return status of blood donors with single reagent reaction to enzyme linked immunosorbent assay(ELISA)and its influencing factors.Methods A total of 291 blood donors who applied to return from January 2018 to December 2023 in Yangzhou Blood Center were selected as research objects,and were divided into returning group(n=186)and shielding group(n=105)according to whether they were successfully returned.The causes of unqualified blood donors were analyzed,and the influencing factors of blood donors with single reagent reaction were analyzed by multivariate Logistic regression.Results The number of hepatitis B two-for-half unqualified blood donors was the largest,accounting for 69.52％,followed by treponema pallidum particle agglutination assay,accounting for 7.62％.The proportion of repeated blood donation,short interval of blood donation,whole blood donation and vaccination of hepatitis B vaccine in returning group were significantly higher than those in shielding group(P＜0.05).Multivariate Logistic regression analysis showed that the number of blood donation,interval time,type of blood donation and vaccination of hepatitis B vaccine were independent factors influencing the return of single reagent reactive blood donors(P＜0.05).Conclusion In Yangzhou area,the probability of successful return is higher in the single reagent reactive blood donors with short interval of blood donation,more blood donation times,whole blood donation type and vaccination of hepatitis B vaccine.

Objective To investigate the effect of rehabilitation instruction based on resourceful theory on the execution intention of exercise behaviour in patient with the first-episode stroke.Methods From September 2022 to February 2023,a total of 80 inpatients with first-episode stroke from a general hospital in Anhui Province participated in the study.Patients in Wards Ⅰ and Ⅱ were assigned to an intervention group,while those in Wards Ⅲ and Ⅳ were assigned to a control group,with 40 patients per group.The patients in control group received standard interventions,while those in the intervention group were offered with a rehabilitation instruction based on resourceful theory in addition to the standard interventions.Stroke patient rehabilitation exercise behaviour execution intention questionnaire,resourcefulness scale and modified Barthel were employed for assessments before and after the intervention.Results After the intervention,the intervention group showed significantly higher scores inexecutive intention of exercise behaviour,resourcefulness level and daily living activities compared to those in the control group(all P＜0.01).Conclusion Rehabilitation instruction based on the theory of resourcefulness can effectively increase the executive intention of exercise behaviour in the patients with first-episode stroke,improve the resourcefulness thinking levels and enhance the daily living activity of the patients.

Objective:To study the current status of longitudinal extrauterine growth restriction (EUGR) in extremely preterm infants (EPIs) and to develop a prediction model based on clinical data from multiple NICUs.Methods:From January 2017 to December 2018, EPIs admitted to 32 NICUs in North China were retrospectively studied. Their general conditions, nutritional support, complications during hospitalization and weight changes were reviewed. Weight loss between birth and discharge > 1SD was defined as longitudinal EUGR. The EPIs were assigned into longitudinal EUGR group and non-EUGR group and their nutritional support and weight changes were compared. The EPIs were randomly assigned into the training dataset and the validation dataset with a ratio of 7∶3. Univariate Cox regression analysis and multiple regression analysis were used in the training dataset to select the independent predictive factors. The best-fitting Nomogram model predicting longitudinal EUGR was established based on Akaike Information Criterion. The model was evaluated for discrimination efficacy, calibration and clinical decision curve analysis.Results:A total of 436 EPIs were included in this study, with a mean gestational age of (26.9±0.9) weeks and a birth weight of (989±171) g. The incidence of longitudinal EUGR was 82.3%(359/436). Seven variables (birth weight Z-score, weight loss, weight growth velocity, the proportion of breast milk ≥75% within 3 d before discharge, invasive mechanical ventilation ≥7 d, maternal antenatal corticosteroids use and bronchopulmonary dysplasia) were selected to establish the prediction model. The area under the receiver operating characteristic curve of the training dataset and the validation dataset were 0.870 (95% CI 0.820-0.920) and 0.879 (95% CI 0.815-0.942), suggesting good discrimination efficacy. The calibration curve indicated a good fit of the model ( P>0.05). The decision curve analysis showed positive net benefits at all thresholds. Conclusions:Currently, EPIs have a high incidence of longitudinal EUGR. The prediction model is helpful for early identification and intervention for EPIs with higher risks of longitudinal EUGR. It is necessary to expand the sample size and conduct prospective studies to optimize and validate the prediction model in the future.

Objective:To observe the clinical efficacy of filiform needle combined with fire needles for cervical radiculopathy(CR)due to wind-cold obstructing the meridians. Methods:A total of 60 patients with CR due to wind-cold obstructing the meridians were randomized into an observation group and a control group,with 30 cases in each group.The control group was treated with filiform needle treatment,and the observation group was treated with additional fire needle point-pricking treatment.The visual analog scale(VAS)and 20-point scale of CR developed by Yasuhisa TANAKA(YT-20)were scored before treatment and after 1,2,3,and 4 weeks of treatments.The tenderness threshold was measured.The local skin temperature was measured by an infrared thermal imager.A safety evaluation was performed after treatment. Results:After treatment,the VAS score in the observation group at each time point was lower than that in the control group(P<0.05).The YT-20 score,tenderness threshold,and local skin temperature in the observation group were all higher than those in the control group(P<0.05).The VAS score in the observation group at each time point after treatment was lower than that before treatment(P<0.05),and the YT-20 score,tenderness threshold,and local skin temperature at each time point were higher than those before treatment(P<0.05).After 3 and 4 weeks of treatments,the VAS score in the control group was lower than that before treatment(P<0.05),and the YT-20 score,tenderness threshold,and local skin temperature were higher than those before treatment(P<0.05).The group factor effects of VAS and YT-20 scores,tenderness threshold,and local skin temperature between the two groups were statistically significant(P<0.05).There was no significant difference in the time effect and the interaction effect between time and group(P>0.05). Conclusion:Filiform needle combined with fire needle or applied alone both can relieve neck pain in patients with CR due to wind-cold obstructing the meridians,and improve the temperature of the neck.The combination of filiform needle and fire needle works more quickly and has better efficacy.

Objective:To investigate the prevalence and predictors of delayed function independence (DFI) in patients with acute vertebrobasilar artery occlusion (VBAO) achieved successful recanalization after endovascular therapy.Methods:Patients with acute VBAO received endovascular treatment in the Departments of Neurology, the First Affiliated Hospital of University of Science and Technology and General Hospital of Eastern Theater Command, PLA from December 2015 to December 2018 were retrospectively enrolled. The demographic, clinical, laboratory and imaging data were collected. Early functional independence (EFI) was defined as the modified Rankin Scale score 0-2 at discharge, and DFI was defined as the modified Rankin Scale score 0-2 at 90 d after discharge for non-EFI patients. Multivariate logistic regression analysis was used to determine the independent predictors of DFI. Results:A total of 122 patients with acute VBAO were included. Their age was 61.8±11.9 years old and 91 (74.6%) were male. The median Glasgow Coma Scale (GCS) score was 7, the median National Institutes of Health Stroke Scale (NIHSS) score was 26.5, and the median posterior circulation Alberta Stroke Program Early CT Score (pc-ASPECTS) score was 9. Twenty-four patients (20.0%) had EFI; of the 98 patients with non-EFI, 18 (18.4%) had DFI. Multivariate logistic regression analysis showed that male (odds ratio [ OR] 0.038, 95% confidence interval [ CI] 0.002-0.658; P=0.025), cardiogenic embolism ( OR 0.116, 95% CI 0.023-0.579; P=0.009), baseline NIHSS score ( OR 1.136, 95% CI 1.040-1.242; P=0.005) and lung infection ( OR 6.089, 95% CI 1.451-25.562; P=0.014) were the independent predictors of DFI. Conclusions:Nearly 1/5 of the non-EFI patients have DFI. Male, cardiogenic embolism, lower baseline NIHSS score and without pulmonary infection are the independent predictors of DFI.

Many efforts have been made to understand excitotoxicity and develop neuroprotectants for the therapy of ischemic stroke. The narrow treatment time window is still to be solved. Given that the ischemic core expanded over days, treatment with an extended time window is anticipated. Bestrophin 1 (BEST1) belongs to a bestrophin family of calcium-activated chloride channels. We revealed an increase in neuronal BEST1 expression and function within the peri-infarct from 8 to 48 h after ischemic stroke in mice. Interfering the protein expression or inhibiting the channel function of BEST1 by genetic manipulation displayed neuroprotective effects and improved motor functional deficits. Using electrophysiological recordings, we demonstrated that extrasynaptic glutamate release through BEST1 channel resulted in delayed excitotoxicity. Finally, we confirmed the therapeutic efficacy of pharmacological inhibition of BEST1 during 6-72 h post-ischemia in rodents. This delayed treatment prevented the expansion of infarct volume and the exacerbation of neurological functions. Our study identifies the glutamate-releasing BEST1 channel as a potential therapeutic target against ischemic stroke with a wide time window.

ObjectiveTo investigate the characteristics of serum perfluoroalkyl substances （PFASs） exposure and potential influencing factors among community residents in Songjiang District， Shanghai. MethodsIn August 2021， residents who underwent routine health checkups in a community in Songjiang District， Shanghai were recruited as study subjects. The inclusion criteria were adult residents who had lived in the area for more than 3 years， had no occupational exposure history， no underlying diseases， were not pregnant， and were able to complete the questionnaire independently and sign the informed consent form. A questionnaire survey was conducted and venous blood samples were collected. The concentrations of 15 PFASs in serum were determined using ultra-high performance liquid chromatography-tandem quadrupole mass spectrometry （UPLC-MS/MS）. Ordered multi-class logistic regression， interquartile range （IQR）， and odds ratio （OR） were used to explore the influencing factors and risk magnitude of serum PFASs concentrations. ResultsOf the 15 PFASs， 14 were detected， and the detection rates of 7 PFASs were higher than 50%. The highest detected concentrations among the PFASs were PFOS （perfluorooctane sulfonate）， PFOA （perfluorooctanoic acid）， and PFHxS （perfluorohexane sulfonate）， with median concentrations of 48.61 μg∙L^-1， 37.29 μg∙L^-1， and 36.51 μg∙L^-1， respectively. The strongest correlation was between PFDA and PFUnDA （r=0.93， P<0.05）， followed by PFOS and PFDA （r=0.86， P<0.05）. Age， frequency of plastic product use， time spent indoors per day， personal annual income， tea consumption， and daily water intake were potential factors for exposure to PFASs. Among them， age was positively correlated with PFASs； tea consumption was positively correlated with PFNA and PFOA； PFHpA was negatively correlated with the frequency of plastic product use and personal annual income； and PFOS was negatively correlated with the time spent indoors per day. ConclusionThe exposure to serum PFASs among community residents in Songjiang District was relatively serious， and the main components were traditional PFOA， PFOS， and PFHx. Different sociodemographic characteristics had varying degrees of influence on the concentrations of PFASs in serum. The impact of PFASs exposure on the health of community residents deserves further investigation.

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.