Objective: To investigate the status of tuberculin skin test (TST) results and the influencing factors among student close contacts with pulmonary tuberculosis, so as to provide the evidence for developing prevention and control strategies for pulmonary tuberculosis among students. Methods: The students aged 15 years and above who had close contact with pulmonary tuberculosis cases in Yuecheng District, Shaoxing City, Zhejiang Province, from October 2016 to December 2023 were recruited and investigated using questionnaires and TST to collect demographic information, contact history, and TST results. The influencing factors for TST positivity among student close contacts with pulmonary tuberculosis were analyzed using a multivariable logistic regression model. Results: A total of 5 507 student close contacts were investigated, including 2 982 males and 2 525 females, with a male-to-female ratio of 1.18∶1. The mean age was (19.10±1.71) years. Among them, 397 (7.21%) were technical secondary school students, 766 (13.91%) were senior high school students, 2 556 (46.41%) were junior college students, and 1 788 (32.47%) were college students or above. A total of 958 students tested positive for TST, with a positivity rate of 17.40%. The rates of general positivity, moderate positivity, and strong positivity were 10.53%, 4.98% and 1.89%, respectively. Multivariable logistic regression analysis showed that senior high school students (OR=1.473, 95%CI: 1.009-2.152) and junior college students (OR=1.467, 95%CI: 1.074-2.005), as well as those with an exposure-to-screening interval of ≥46 days (46-<61 days, OR=2.043, 95%CI: 1.478-2.826; ≥61 days, OR=1.291, 95%CI: 1.018-1.637) had a higher risk of TST positivity. Female student close contacts had a lower risk of TST positivity (OR=0.753, 95%CI: 0.649-0.873). Conclusion The TST positivity rate was relatively high, and gender, school type, and exposure-to-screening interval were influencing factors for TST positivity among student close contacts with pulmonary tuberculosis.

ObjectiveTo analyze the influencing factors for recurrence in successfully treated pulmonary tuberculosis patients with isoniazid-resistant and rifampicin-sensitive in Shaoxing City, Zhejiang Province. MethodsData on general demographic information, treatment information and drug susceptibility test results for pulmonary tuberculosis patients admitted to the designated tuberculosis medical institutions and registered in the tuberculosis information management system was collected in Shaoxing City from January 2011 to August 2024. A total of 428 patients with isoniazid resistance (including isoniazid single resistance and multiple resistance) but who were successfully treated were included in the study. Information for the recurrence after successful treatment of the patients was analyzed. The Cox proportional hazards models were used to analyze the influencing factors of recurrence in patients. ResultsAmong the 428 successfully treated patients included in the study, 31 cases (accounting for 7.24%) had recurrence by the end of the observation period, with a recurrence rate density of 1.31 per 100 person-years and a median recurrence time of 0.99 (0.08, 8.27) years. Among the relapsed population, 51.61% of the patients relapsed within one year after successful treatment. 77.42% of the patients relapsed within two years after successful treatment. Multivariate Cox regression analysis showed that when isoniazid resistance was discovered, the diagnosis classification of relapse (HR=4.115, 95%CI: 1.734‒9.767) and positive 0-month sequence smear (HR=4.457, 95%CI: 1.053‒18.866) were risk factors for recurrence after successful treatment in patients. ConclusionRegular follow-up should be strengthened for at least two years after the successful treatment of isoniazid-resistant pulmonary tuberculosis patients. Special attention should be paid to the treatment effect and regular re-examination and monitoring after the end of the treatment course of isoniazid-resistant pulmonary tuberculosis patients who have been re-treated and were sputum smear positive at baseline, so as to prevent recurrence and disease progression in high-risk populations.

Objective To analyze TCM master Yan Zhenghua's medication rules of ascending,descending,floating and sinking properties for the treatment of respiratory diseases,inherit his valuable academic experience.Methods Using four books edited by Professor Yan Zhenghua's disciples as the main source,Professor Yan Zhenghua's prescriptions for clinical treatment of respiratory diseases were systematically collected and analyzed.Statistical analysis was conducted on the patients'gender and age,differentiation of diseases and syndrome types of the prescriptions,as well as the properties of ascending,descending,floating and sinking,dosage,and commonly used pairs of Chinese materia medica.Results Totally 208 prescriptions were included in this study,involving 178 kinds of Chinese materia medica and 64 kinds of monarch drug,most of them were descending and sinking drug,and the whole prescription was mainly descending and sinking.On average,each prescription used 13.2 kinds of Chinese materia medica,and most dosage points were within the range of conventional dosage.Among later adding medicines,Houttuyniae Herba had the highest frequency of use.The medicinal pairs of Asteris Radix et Rhizoma-Cynanchi Stauntonii Rhizoma et Radix,Cynanchi Stauntonii Rhizoma et Radix-Stemonae Radix and Asteris Radix et Rhizoma-Stemonae Radix were often used.Armeniacae Semen Amarum,Fritillariae Thunbergii Bulbus,and Asteris Radix et Rhizoma were commonly used in those descending and sinking prescriptions.Conclusion In the treatment of respiratory diseases,Professor Yan Zhenghua prefers to use descending and sinking drugs with ascending and floating drugs and dual trend drugs to regulate qi activity,and has the characteristics of Menghe medical school,which is"mild and flexible medication".

Objective To explore the histomorphology and related indicators in a rat model of skeletal muscle injury in an artificial experimental chamber as special environment in Northwest China to stimulate a hot and dry desert environment in order to screen the evaluation indicators of skeletal muscle injury in a hot and dry environment.Methods Eighteen male SD rats (8 weeks old)were randomly divided into blank control group (room temperature,free movement),room temperature exercise group,and hot and dry exercise group,with 6 rats in each group.After the rats were subjected to adaptation training for 3 d (a 3-D exercise treadmill,speed:16 m/min,slope:-16°,time:5～10 min),the rats in the 2 exercise groups were subjected to a treadmill once per day at a speed of 19.3 m/min and a slope of-16°till the end of exhaustion,and those in the hot and dry exercise group were housed in an artificial experimental chamber at a temperature of (40±1 )℃,a humidity of (10±2)％.After 6 d of continuous exercise,the rat soleus muscle tissue was taken to observe the pathological changes,and the serum samples were detected for skeletal muscle injury markers,inflammatory factors,oxidative stress related factors,hormones and other related indicators. Results When compared with the blank control group and the room temperature exercise group,the hot and dry exercise group had significantly higher concentrations of creatine kinase (CK),creatine kinase isoenzyme (CK-MB)and muscle-specific creatine kinase (CK-MM)(P<0.05 ),elevated contents of TNF-α,IL-1βand IL-6 (P<0.05),raised malondialdehyde (MDA)content and lower superoxide dismutase (SOD)activity (P<0.05 ),lower testosterone (T)but higher corticosterone (Cort)levels and thus lower testosterone/corticosterone (T/Cort)ratio (P<0.05 ).The markers of skeletal muscle injury,CK,CK-MB and CK-MM were positively correlated with the pro-inflammatory cytokines,TNF-α,IL-1βand IL-6 (P<0.05),positively with the antioxidant indicator MDA but negatively with SOD (P<0.05 ),and positively correlated with Cort level and negatively with T level and T/Cort ratio (P<0.05).Conclusion A stable rat model of skeletal muscle injury in a hot and dry desert environment is successfully constructed.The severity of skeletal muscle injury is greater in rats in a hot and dry desert environment than those in a normal temperature environment. TNF-α and IL-1β are found to be indirect indicators to evaluate skeletal muscle injury in the environment.

Objective To investigate the beneficial effect of Kai-Xin-San combined with fluoxetine in improving depression-like behaviors on chronic unpredictable mild stress(CUMS)induced depression model mice.Methods The present study aimed to assess the potential of Kai-Xin-San in combination with fluoxetine to ameliorate depression-like behaviors in a CUMS induced mouse depression model.Behavioral tests,such as the sucrose preference test were employed to evaluate the efficacy of the treatment.Additionally,the levels of suppressed stress factors were measured using the ELISA method.The morphology of hippocampal tissue was evaluated using the HE staining method,Nissl Staining and TUNEL staining methods.Furthermore,western blotting analysis was utilized to determine the expression levels of proteins such as Caspase-3,and Caspase-9.Results The co-administration of Kai-Xin-San and fluoxetine resulted in a significant increase in sucrose preference rate in model mice.This effect was comparable to that of fluoxetine alone at the standard clinical dose.Furthermore,the combination treatment up-regulated the levels of suppressed stress factors,reduced the apoptosis of hippocampus induced by depression and regulated the apoptosis signaling pathway in hippocampus.Conclusion The combination of Kai-Xin-San and fluoxetine has been shown to be an effective treatment for depression-like behavior in animal models,resulting in a reduction in the required clinical dosage of fluoxetine.This effect may be attributed to the up-regulation of neurotransmitter expression,inhibition of stress axis activation,and central nervous inflammation.

Objective:To investigate the types, incidences, and clinical characteristics of shock in polytrauma patients at different stages after polytrauma.Methods:A retrospective study was conducted on polytrauma patients admitted to multiple trauma centers from June 2020 to December 2021. The inclusion criteria were patients >18 years old and treated due to polytrauma. Exclusion criteria included an admission time of more than 48 h after trauma, a history of malignancy, or metabolic, consumptive, and immunological diseases. The early stage was defined as the period of ≤48 h after polytrauma, and the middle stage was defined as the period between 48 h and 14 days. The patient’s medical history, clinical manifestations, laboratory tests, imaging examination, injury severity score (ISS), and Glasgow coma scale (GCS) were collected. The types, incidences, and clinical characteristics of shock in different stages after polytrauma were analyzed, according to the diagnostic criteria of each type of shock. The differences between the groups were compared by Student’s t test, χ2 test or Mann-Whitney U test. Results:The incidence of the early and middle stage shock after polytrauma were 73.1% and 36.4%, respectively, with statistically significant difference between stages ( P<0.01). There were significant differences in the incidence of hypovolemic shock (83.6% vs. 28.4%), distributed shock (13.7% vs. 80.9%) and cardiogenic shock (3.5% vs. 6.6%) between stages (all P<0.05). The incidence of obstructive shock (8.4% vs. 9.7%, P>0.05) was similar between stages. The incidence of undifferentiated shock was 1.6% and 1.2%, respectively. There were 9.5% patients with multifactorial shock in the early stage and 14.4% in the middle stage. Totally 7 combinations of multifactorial shock were found in different stages after polytrauma. In the early stage, the combination of HS and DS accounted the highest ratio (42.3%) and followed by HS and OS for 28.8%. In the middle stage, the combination of HS and DS was the most common (48.6%) and followed by DS and OS (24.3%). Conclusions:The incidence of shock in polytrauma patients is high. Different types of shock can occur simultaneously or sequentially. Therefore a comprehensive resuscitation strategy is significant to improve the success rate of treatment.

Pigs are considered as ideal donors for xenotransplantation because they have many physiological and anatomical characteristics similar to human beings. However, antibody-mediated immunity, which includes both natural and induced antibody responses, is a major challenge for the success of pig-to-primate xenotransplantation. Various genetic modification methods help to tailor pigs to be appropriate donors for xenotransplantation. In this study, we applied transcription activator-like effector nuclease (TALEN) to knock out the porcine α-1, 3-galactosyltransferase gene GGTA1, which encodes Gal epitopes that induce hyperacute immune rejection in pig-to-human xenotransplantation. Meanwhile, human leukocyte antigen-G5 gene HLA-G5, which acts as an immunosuppressive factor, was co-transfected with TALEN into porcine fetal fibroblasts. The cell colonies of GGTA1 biallelic knockout with positive transgene for HLA-G5 were chosen as nuclear donors to generate genetic modified piglets through a single round of somatic cell nuclear transfer. As a result, we successfully obtained 20 modified piglets that were positive for GGTA1 knockout (GTKO) and half of them expressed the HLA-G5 protein. Gal epitopes on the cell membrane of GTKO/HLA-G5 piglets were completely absent. Western blotting and immunofluorescence showed that HLA-G5 was expressed in the modified piglets. Functionally, the fibroblasts from the GTKO/HLA-G5 piglets showed enhanced resistance to complement-mediated lysis ability compared with those from GTKO-only or wild-type pigs. These results indicate that the GTKO/HLA-G5 pigs could be a valuable donor model to facilitate laboratory studies and clinics for xenotransplantation.
Animals ; Animals, Genetically Modified ; Gene Knockout Techniques ; HLA Antigens ; Humans ; Nuclear Transfer Techniques ; Swine ; Transplantation, Heterologous

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system is a hotspot of gene editing and gene expression research, in which CRISPR/Cas13 system provides a new direction for RNA interference and editing. In this study, we designed and synthesized the corresponding gRNAs of CRISPR/Cas13a and CRISPR/Cas13b systems in non-homologous end joining (NHEJ) pathway, such as Ku70 and Lig4, and then detected the expression of ku70 and lig4 in HEK293T cells. The CRISPR/Cas13a system could efficiently knockdown the mRNA expression of ku70 and lig4 more than 50%, and CRISPR/Cas13b system also suppressed ku70 and lig4 about 92% and 76%, respectively. Also, CRISPR/Cas13a, b systems could down-regulate Ku70 and Lig4 proteins level to 68% and 53%, respectively. The study demonstrates that the CRISPR/Cas13 system could effectively knockdown the expression of RNA and protein in HEK293T cells, providing a new strategy for gene function and regulation research.
CRISPR-Cas Systems ; DNA Ligase ATP ; genetics ; Gene Expression Regulation ; genetics ; Gene Knockdown Techniques ; HEK293 Cells ; Humans ; Ku Autoantigen ; genetics

Objective:To compare ultrasound (US) guided versus computed tomography (CT) guided radiofrequency ablation (RFA) in treatment of early hepatocellular carcinoma (HCC).Methods:The data of 133 patients with early HCC treated by RFA in the Department of Hepatobiliary Surgery of Shandong Provincial Hospital from February 1, 2015, to January 31, 2017, was analyzed retrospectively. These patients were divided into two groups: the US-guided group and the CT-guided group. The clinical data was collected and the factors affecting prognosis were analyzed.Results:Compared with the CT-guided group, the operation time of the US-guided group was significantly shorter [(29.0±12.0)min vs. (55.0±19.0)min, P<0.05], but the number of ablation sessions per tumor was significantly less [(1.1±0.3) vs. (2.0±0.6), P<0.05]. There was no significant difference in the complete ablation rates, postoperative complication rates and postoperative length of hospital stay between the two groups ( P>0.05). The CT-guided group was superior to the US-guided group in the local tumor recurrence and progression-free survival rates ( P<0.05). On multivariate analysis, CT-guided RFA was an independent protective factor for local tumor recurrence ( HR=0.266, 95% CI: 0.073-0.967, P<0.05) and progression-free survival ( HR=0.415. 95% CI: 0.213-0.806, P<0.05), while AFP >20 ng/ml ( HR=4.821, 95% CI: 1.714-13.560, P<0.05) was an independent risk factor for progression-free survival. Conclusion:CT-guided percutaneous RFA was superior to US-guided RFA in local treatment of early HCC, probably related to more needle placements and longer ablation time under CT guidance.