OBJECTIVE:To study the effect of alcohol extract of Toddalia asiatica on the inflammation-associated cytokines of model rats with adjuvant arthritis(AA). METHODS:70 SD rats were randomly divided into a normal control group,a model con-trol group,a positive control chemical medicine group(Leflunomide tablets,0.012 g/kg),a positive control TCM group(Tripter-ygium glycosides tablets,0.012 g/kg)and the groups of low,medium and high-dose [1,4,6 g(crude drug)/kg] alcohol extract of T. asiatica,with 10 rats in each group. The rats in all groups except for the normal control group were given complete Freund’s complete adjuvant id for the establishment of AA models. At the same time,the rats in the drug administration groups were given corresponding drugs ig,while those in the normal control group and the model control group were given isometric normal saline ig,twice a day,for 28 consecutive days. The degree of toe swelling,arthritis index and the levels of interleukin(IL)-1β,IL-6, IL-10,tumor necrosis factor α(TNF-α)and prostaglandin E2(PGE2)in serum and synovial membranes of all groups of rats were determined. RESULTS:Compared to the normal control group,the model control group demonstrated higher degree of primary and secondary toe swelling,arthritis index and levels of IL-1β,IL-6,TNF-αand PGE2 in serum and joint synovial membrane,and low-er level of IL-10 therein(P<0.01). Compared to the model control group,all the above-mentioned indexes of the rats in drug ad-ministration groups significantly improved(P<0.05 or P<0.01). CONCLUSIONS:The alcohol extract of T. asiatica. has a preven-tive and therapeutic effect on the model rats with AA by regulating the expression of anti-inflammatory and proinflammatory cyto-kines in serum and synovial membrane.

Objective To investigate the possibility of utilizing the mixed bacteria liquid and the Escherichia coli ( E.coli) liquid to establish the chronic salpingitis model of New Zealand rabbits, respectively.Methods Taken as the study object, the un-pregnant New Zealand rabbits (4~5 years old) were randomly divided into three groups: the normal group, the mixed bacteria experimental group and the E.coli experimental group.The trans-vaginal intrauterine intubation operation was performed for the injection of the mixed bacteria liquid and the E.coli liquid.Visual observation was to evaluate the gross pathological changes of the salpingitis and the pelvic cavity.HE staining and the light microscope were used to observe the micro-pathological changes of salpingitis.Results On the 15th day after modeling, increased pelvic effusion, dense peritoneal adhesion, interstitial hyperplasia and infiltration of lymphocytes were observed in both experimental groups. Conclusion Through the trans-vaginal intrauterine intubation operation, the chronic salpingitis model of New Zealand rabbits could be successfully established either by using the mixed bacteria liquid or by using the E.coli liquid.

ObjectiveTo explore the childhood trauma experience and its relation to the eye emotion recognition ability in patients with schizophrenia.MethodsSeventy-one schizophrenic patients and fifty-two healthy subjects were assessed with the Childhood Trauma Questionnaire (CTQ) and the Eye Basic Emotion Discrimination Task (EBEDT).ResultsThe CTQ total score in the patients group was significantly higher than those in the control group (47.86 ± 13.26 vs 34.27 ± 7.27,P ＜ 0.01 ).The subscores in emotional abuse,physical abuse,sexual abuse,emotional neglect and physical neglect of CTQ in the patients group were also significantly higher than those in the control group.(all P＜ 0.01 ).Compared with controls,the patients had significantly lower total accuracy( (86.74 ± 6.16vs73.42 ± 11.86) ％,P ＜ 0.01 ) and longer total reaction time ( (2776.75 ± 703.42 vs 4321.40 ± 1956.83 )ms,P ＜ 0.01 )of EBEDT.The significant difference were also found on the accuracy and reaction time of the EBEDT six basic emotion between the two groups ( all P ＜ 0.01 ).Correlation analysis indicated that CTQ total score were negatively correlated with the EBEDT total accuracy ( r=-0.390,P＜0.01 ),and its sub-accuracy of happiness,surprise,sadness,and anger ( respectively r =- 0.419,r =- 0.451,r =- 0.391,r =- 0.344,P ＜ 0.01 ) in the patients group.The correlation between CTQ total score and EBEDT total accuracy was also found in the control group.ConclusionChildhood trauma is related to eye basic emotion recognition in schizophrenia patients,suggesting that the trauma during childhood can affect the social recognition ability.

Objective:To study the incidences of complications in twins with birth weight discordance (BWD) during hospitalization.Methods:From January 2011 to December 2020, twins born in the Department of Obstetrics and hospitalized in NICU of our hospital were retrospectively studied. Twins with BWD>15% were assigned into BWDT group and BWD≤15% into twins with birth weight concordant (BWCT) group. Complications during hospitalization were compared between the two groups.Results:A total of 1 546 pairs of twins were enrolled, including 486 (31.4%) in BWDT group and 1 060 (68.6%) in BWCT group. Compared with BWCT group, BWDT group had significantly higher incidences of pulmonary surfactant (PS) utilization (47.7% vs. 42.2%), continuous positive airway pressure ≥ 24 h (40.5% vs. 35.0%), high-flow nasal cannula ≥ 24 h (22.8% vs. 16.3%), neonatal respiratory distress syndrome (52.9% vs. 47.1%), bronchopulmonary dysplasia (BPD) (15.6% vs. 11.1%), persistent pulmonary hypertension of the newborn (3.1% vs. 1.4%) and anemia (79.6% vs. 70.1%) (all P<0.05). After adjusting for confounding factors, the risks of pulmonary hemorrhage ( OR=2.036, 95% CI 1.119-3.703, P=0.020) and BPD ( OR=2.960, 95% CI 1.656-5.219, P=0.010) in BWDT group were higher than BWCT group. Conclusions:BWD twins has higher incidences of complications during hospitalization than BWC twins.

ObjectiveTo explore the association between triglyceride-glucose （TyG） index and major adverse cardiovascular events （MACEs） risk in coronary heart disease （CHD） patients with blood stasis syndrome after percutaneous coronary intervention （PCI）. MethodsA total of 857 CHD patients with blood stasis syndrome after PCI were enrolled and divided into four groups according to the baseline TyG index quartiles， Q1 （TyG ＜ 8.51）， Q2 （8.51 ≤ TyG ＜ 8.88）， Q3 （8.88 ≤ TyG ＜ 9.22）， and Q4 （TyG ≥ 9.22）. The clinical outcome was defined as a compound endpoint of cardiovascular events including cardiac death， non-fatal myocardial infarction， unplanned revascularization， in-stent restenosis and stroke. The machine learning Boruta algorithm was used for feature selection related to MACEs risk. Kaplan-Meier survival analysis and Cox proportional hazards regression model were used to compare the differences in MACEs risk among the four groups. Restricted cubic spline （RCS） and subgroup analysis were performed to determine the relationship between the TyG index and MACEs risk. The area under the receiver operating characteristic （ROC） curve （AUC）， calibration curve and Hosmer-Lemeshow test， and decision curve analysis （DCA） were plotted to evaluate the predictive value of the TyG index for MACEs risk. ResultsThe median follow-up time of the included patients was 2.45 years. During the follow-up period， 313 cases （36.52%） of new MACEs occurred. The incidence of MACEs in Q1， Q2， Q3， Q4 group was 28.17% （60/213）， 29.05% （61/210）， 39.45% （86/218） and 49.07% （106/216）， respectively. Kaplan-Meier survival analysis suggested statistically significant differences in MACEs risk among the four groups （P＜0.001）. Cox proportional hazards regression model analysis found that the risk of MACEs in patients with high TyG index increased by 60.1% （P＜0.01）. Using Q1 as the reference， the MACEs risk in Q2， Q3 and Q4 groups gradually increased， and the trend was statistically significant （P＜0.05）. RCS model suggested that the TyG index was nonlinearly associated with the MACEs risk （P＜0.001）. The TyG index had a good predictive performance for MACEs risk according to ROC analysis （AUC=0.758， 0.724-0.792） and Hosmer-Lemeshow test （χ² = 4.319， P = 0.827）. Additionally， DCA analysis also suggested a good clinical efficacy of the TyG index for predicting MACEs. Subgroup analysis showed that different baseline TyG index was positively correlated with the MACEs risk in the stratification of age， male， BMI， history of diabetes and hypertension， and low-density lipoprotein cholesterol （LDL-C）≥1.8 mmol·L^-1（P＜0.05）. ConclusionThe elevated TyG index is closely corrected with an increased risk of MACEs in CHD patients with blood stasis syndrome after PCI， indicating a guiding significance for early risk stratification and clinical decisions.

Nerve growth factor (NGF) can promote the development, differentiation and regeneration of neurons. Recently, in order to efficiently produce human NGF (hNGF) drugs with better efficacy, we created transgenic mice expressing hNGF specifically in their salivary glands, and purified highly active hNGF protein from their saliva. Some studies reported that the NGF secretion in mouse saliva is affected by gender and age. Here, in order to select hNGF transgenic mice with high NGF secretion for saliva collection and hNGF purification, we divided transgenic mice into 4 groups, including 28-day-old young males and females, 63-day-old adult males and females. We compared their saliva volume, total salivary protein amount, salivary mNGF protein amount and salivary hNGF protein amount. The results showed that the saliva volume as well as amounts of total salivary protein, salivary mNGF protein and salivary hNGF protein secreted by 63-day-old transgenic mice were significantly higher than those secreted by sex-match 28-day-old transgenic mice, and the salivary hNGF protein amount secreted by male transgenic mice at the age of 63 days was significantly higher than that of female transgenic mice at the same age; Among 4 groups of mice, 63-day-old male transgenic mice secreted the highest salivary hNGF content, which was about 46 times higher than that secreted by the 28-day-old female transgenic mice. Therefore, 63-day-old male transgenic mice should be selected for saliva collection and hNGF purification.
Animals ; Cell Differentiation ; Female ; Humans ; Male ; Mice ; Mice, Transgenic ; Nerve Growth Factor ; Saliva

[Abstract] Objective: To screen candidate epitopes of breast cancer HLA-A2 restrictive neoantigen and to identify high frequency mutation sites in breast cancer neoantigen by using bioinformatics method. Methods: NCBI and GDC databases were used to search missense mutation sites formed by single nucleotide mutation in breast cancer among reported literatures and sequencing data. The new antigen epitopes were predicted by HLA-A2 antigen epitope prediction website BIMAS, SYFPEITHI and artificial neural networkbased NetMHC4.0, and the epitopes with TAP binding power less than Intermediate were eliminated. The candidate epitopes were prioritized by mutation frequency and prediction results. Results: A total of 17 high-frequency mutation genes, including BTLA, ERBB2 and NBPF12 etc, were screened by the above-mentioned methods, and a total of 26 neoantigen epitopes were identified. The binding power of epitopes predicted using BIMAS and SYFPEITHI showed great difference (P<0.05), epitopes in high priority as GSTP1 (A114V , mutation frequency of 5.94%) and BRCA2 (N991H, mutation frequency of 5.40%) etc, were expected to be candidate neo-antigen epitopes; however, their mutation frequency was relatively too low to achieve“universal use” . The possibility of these epitopes used as general breast cancer neo-antigen epitopes is less likely. Conclusion: The common mutation frequency of breast cancer is lower than that of other tumors; it ’s difficult to find“universal”new antigen epitopes of breast cancer; the individualized neoantigen vaccine may be of more promise, which needs further research.

OBJECTIVE To systematically evaluate the efficacy and safety of Shenfu injection combined with chemical medicine in the treatment of coronary heart disease combined with heart failure. METHODS Retrieved from CNKI， CBM， VIP， Wanfang， PubMed， Embase and the Cochrane Library， randomized controlled trials （RCTs） about Shenfu injection combined with chemical medicine （trial group） versus chemical medicine （control group） in the treatment of heart failure with coronary heart disease were collected during the inception to August 2022. After literature screening and data extraction， the qualities of included literature were evaluated and rated by using Cochrane manual and GRADE system. Meta-analysis and Egger’s were performed with RevMan 5.3 software， and TSA 0.9.5.10 Beta software was used for trial sequential analysis. RESULTS Seventeen studies were included， with a total sample of 1 355 patients. The quality grade evidence of GRADE was all low. Meta-analysis showed that cardiac function efficacy [RR＝1.23， 95%CI （1.16，1.30）， P＜0.000 01]， the decrease of brain natriuretic peptide [MD＝－96.06， 95%CI （－116.47， －75.64）， P＜0.000 01] and the increase of left ventricular ejection fraction [MD＝5.32， 95%CI （4.03，6.60）， P＜0.000 01] in trial group were significantly better than control group； there was no statistical significance in the incidence of ADR between 2 groups [RR＝0.52，95%CI（0.22，1.22），P＝0.13]. The results of sequential analysis showed that the sample size included in this study met the requirements of meta-analysis； the results of Egger’s test showed that the results were robust and publication bias had no significant effect on the results. CONCLUSIONS Shenfu injection combined with chemical medicine in the treatment of coronary heart disease combined with heart failure can further improve the clinical symptoms and related indicators， and no serious adverse reaction is observed.

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

To analyze the mid-long-term outcomes of surgical balloon valvuloplasty (SBV) for right ventricular decompression in the treatment of pulmonary atresia with intact ventricular septum (PA/IVS). Methods    Clinical data of consecutive 91 patients who were diagnosed with PA/IVS and underwent SBV in our institution from January 2005 to December 2017 were retrospectively analyzed, including 52 (57.1%) males and 39 (42.9%) females. The median age was 3 months (1 d, 24 months) and the median weight was 4.1 (2.5, 12.0) kg. Results    The SBV was performed in all patients, and 62 of whom received other simultaneous surgeries, including ligation of patent ductus arteriosus (PDA, 33 patients), ligation of PDA with modified Blalock-Taussig shunt (23 patients), ligation of PDA with bidirectional Glenn shunt (6 patients). There was no early postoperative death. The median follow-up time was 8.8 (2.5, 13.4) years, 4 patients were lost. There were 7 (8.0%) deaths and 1 (1.1%) patient with a re-SBV for pulmonary stenosis. The one and a half ventricular repair was performed in 5 (5.7%) patients and Fontan procedure in 2 (2.3%) patients. In addition, the mean Z-value of tricuspid valve annulus was −1.7±1.5, which was significant bigger than that before the operation (t=5.587, P<0.001). Conclusion    SBV via right ventricular outflow tract for right ventricular decompression in the treatment of PA/IVS is safe and reliable. The majority of patients can receive biventricular repair instead of single ventricular palliation by SBV with individually customized shunt.