AIM: To investigate the effect of ligustrazine (Lig) on cerebral injury in LPS-induced septic shock rats and to explore the underlying mechanism.METHODS: Wistar rats (n =48) were randomly divided into control group, LPS group and LPS +Lig treatment group.The rats in LPS group were randomly divided into 2 subgroups at time points of 6 h and 12 h.After ligustrazine treatment, the venous blood was collected by removal of eyeballs to detect the con-centration of neuron-specific enolase (NSE) using ELISA.The nitric oxide (NO) concentration in the homogenate of brain tissues was examined.The apoptosis in the hippocampus was analyzed by TUNEL staining.The protein expression of Bax and Bcl-2 was determined by Western blot.RESULTS: Ligustrazine inhibited the elevation of NSE and NO concentrations in LPS-induced septic shock rats.Furthermore, ligustrazine administration also attenuated LPS-induced increase in Bax ex-pression and decrease in Bcl-2 expression.CONCLUSION: Ligustrazine decreases the concentration of NSE and NO, and attenuates cerebral injury in LPS-induced septic shock rats.These effects may be related to the regulation of Bax and Bcl-2 expression.

ObjectiveTo investigate the association between nucleotide polymorphisms (SNP) of rs2282430 and rs2031234 inSLC26A9 gene and clinical characteristics of asthma in Han children in central China.MethodsA case-control study was performed. Two hundreds and three children with asthma were recruited in this study and 221 normal children were selected as controls. The genotypes of two SNPs inSLC26A9 gene were examined using PCR-RFLP.ResultsBetween children with asthma and controls, the distribution of three genotypes (AA, AG and GG) in rs2282430 locus had signiifcant difference (P=0.042). The percentage of AA genotype was higher in children with asthma than that in controls. In implicit mode (AAvs. AG+GG), the two groups was statistically signiifcant difference (P=0.028). The frequency of A allele was higher in children with asthma than that in controls (P=0.011). Between children with asthma and controls, the distribution of three genotypes (TT, GT, and GG) in rs12031234 locus had no signiifcant difference (P=0.479). The frequency of alleles in rs12031234 locus also had no signiifcant difference (P=0.215). Among asthmatic children with different genotype of rs2282430, the lymphocytecounts (LYM), C-reaction protein (CRP), IgE, neutrophils (NEU%), and eosinophils (EOS%) were not signiifcantly different (P>0.05). Conclu-sionsThe rs2282430 polymorphism inSLC26A9 gene is associated with childhood asthma in the central China and the A allele is the risk factor. The rs2282430 polymorphism is not associated with LYM counts, CRP level, IgE level, NEU%, and EOS%.

To investigate the efficacy and adverse effects of venlafaxine in the treatment of depression, patients were given venlafaxine or amitriptyline for 6 weeks. The efficacy of the drugs were assessed by using HAMD and HAMA at the 4th, 7th, 14th and 42nd day before and after the medication. The adverse effects were evaluated by TESS. Our results showed that the effect time of venlafaxine was (8.52±4.47) d, which was fasler the that of amitriptyline. The effective rate of venlafaxine was 83%, which was faster the that of amitriptyline. but venlafaxine caused less dry mouth, constipation, blurred vusion, tachycardia and anorexia etc. It is concluded that venlafaxine worked faster with less side effect in the treatment of depression as compared with amitriptyline.

[ABSTRACT]OBJECTIVETo investigate the effect of accurate airway humidification on hemorrhage, pharyngalgia, mucosal edema and sputum viscosity in patients with low-temperature plasma coblation-assisted tonsillectomy.METHODS58 cases were divided into three groups by using random numbers.In accurate airway humidification group, atomizing inhalation was carried out by AIRVOTM series apparatus; in oxygen atomizing group, budesonide suspension was used; in control group, saline was used. We evaluated the hemorrhage, pharyngalgia, mucosal edema and sputum viscosity in 3 consecutive postoperative days.RESULTSPharyngalgia in accurate airway humidification group and in oxygen atomizing group were both significantly reduced than that of the control group (P<0.001). Besides, in accurate airway humidification group, mucosal edema and sputum viscosity were significantly improved than that of the oxygen atomizing group (P<0.05) and control group (P<0.05).CONCLUSIONAccurate airway humidification could reduce the complications such as pharyngalgia, mucosal edema and purulent sputum after low-temperature plasma coblation-assisted tonsillectomy, and could accelerate recovery from surgery.

Objective To assess the relationship between maternal plasma tumor necrosis factor-related apoptosis-inducing ligand ( TRAIL ) before 20 weeks′gestation and hypertensive disorder complicating pregnancy (HDCP);and to evaluate the predictive value of plasma TRAIL for HDCP.Methods A 2-phase screening/validation study was designed.In the screening phase , a nested, case-controlled study was performed , the plasma samples collected before 20 weeks′gestation from 20 women who later developed HDCP and 20 age-and gestation week-matched controls were tested in prospective screening test for protein expression profiling during pregnancy and HDCP.Plasma samples were analyzed by a human protein microarray technology designed to detect 507 proteins simultaneously.Differently expressed proteins′functional annotation and clustering were performed by using of Database for Annotation , Visualization and Integrated Discovery ( DAVID) and Gene Ontology ( GO) database.The TRAIL level of plasma samples obtained before 20 weeks′gestation from 53 women who later developed HDCP and 106 similarly matched controls were further validated by ELISA and 62 clinical risk factors were investigated.Logistic regression and ROC analysis were used to evaluate the relationship between TRAIL and HDCP and its predictive value for HDCP.Results In protein microarray analysis , 23 proteins expressed differently before 20 weeks′gestation between the two groups.Further validation results showed that TRAIL levels in HDCP patients were lower significantly (45.7 ±13.1) pg/ml than those in healthy pregnant controls (51.2 ±14.7)pg/ml, P=0.021.Multiple factor logistic regression analysis of 159 pregnancies showed that three features were finally entering the logistic model, they were:anemia (OR=4.87, 95% CI 1.05-24.26), pre-pregnancy BMI (OR=1.72, 95% CI 1.35 -2.19) and TRAIL (OR=0.96, 95% CI 0.92 -0.99).The predictive accuracy of logistic model was 81.8%.The model significantly increases the predictive value (AUC=0.81, 95%CI 0.73-0.87) compared to TRAIL as independent predictor (AUC=0.59, 95%CI 0.51-0.67).Conclusions Totally 23 proteins were expressed differentially before 20 weeks′gestation in plasma of women who later developed HDCP , confirming that HDCP is a heterogeneous disease with different biological changes.The data suggests that plasma TRAIL levels relate with the development of HDCP and its combination with pre-pregnancy BMI and anemia have a high predictive value for HDCP before 20 weeks′gestation.

BACKGROUND:As current studies on isolation, culture andcryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future.OBJECTIVE:To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs.METHODS:Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs.RESULTS AND CONCLUSION:(1) The optimal hAECs separation conditions were as follows:trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.

Objective:Immunoregulation study of umbilical mesenchymal stem cell (UCMSCs) on allogeneic umbilical cord blood(UCB) CD4+T lymphocytes,which proliferation,apoptosis and the differentiation to CD4+CD25+ regulatory T cell (Treg) in vitro. Methods:Establishing on direct contact or transwell co-culture system,adopt in different proportion of UCMCs with phytohaemag-glutinin (PHA)-activated UCB CD4+T lymphocytes were co-cultured. The proliferation of lymphocyte,percent of CD4+CD25+/CD4+and Foxp3 expression, regulatory T cell marker gene were measured. Apoptosis of CD4+T lymphocytes were observed in the direct contact or transwell coculture system of UCMSCs with desamethason( DXM)-stimulated UCB CD4+T lymphocytes. Results: The UCB CD4+T lymphocytes cocultured with UCMSCs with PHA-activating for 3 days,compared with the UCMSCs free control group,the amount of cells was reduced noticeably(P<0. 05) and the percent of CD4+CD25+in CD4+T lymphocytes and Foxp3 expression significantly in-creased(P<0. 01) in a dose dependent way(P<0. 05). The UCB CD4+T lymphocytes cocultured with UCMSCs with DXM-inducing for 7 days,the apoptosis rate was significantly lower than that of the control group without UCMSCs (P<0. 01). These effects were partially attenuated in transwell coculture but could not be eliminated. Conclusion: UCMSCs are negative effect on UCB CD4+T lymphocytes-mediated immunity effects,and mainly manifested in the regulation on cell proliferate ability and differentiation rather than promoting apoptosis.

Objective To analyze the application value of cervical vascular color Doppler ultrasound com-bined with serum microRNA-222-3p(miR-222-3p)and microRNA-206(miR-206)in the diagnosis of carotid artery stenosis in patients with cerebral infarction.Methods A total of 90 patients with cerebral infarction ad-mitted to this hospital from July 2018 to July 2022 were selected as the study group,and were divided into mild stenosis group(29 cases),moderate stenosis group(35 cases),and severe stenosis group(26 cases)ac-cording to the degree of carotid artery stenosis.In addition,96 healthy people who underwent physical exami-nation in the hospital were selected as the control group.Real-time fluorescence quantitative PCR was used to detect the levels of serum miR-222-3p and miR-206 in each group.All patients with cerebral infarction under-went cervical vascular color Doppler ultrasound examination,and the clinical data of patients with different de-grees of stenosis were analyzed.Multivariate Logistic regression was used to analyze the factors affecting the degree of carotid artery stenosis in patients with cerebral infarction.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic value of carotid ultrasound indicators,serum miR-222-3p and miR-206 levels for the degree of carotid artery stenosis in patients with cerebral infarction.Results Compared with the control group,the study group had a significant increase in the serum level of miR-222-3p and a sig-nificant reduction in the serum level of miR-206(P＜0.05).Compared with the mild stenosis group,the peak systolic velocity(PSV),end diastolic volume(EDV)and serum miR-222-3p level were increased,and the ser-um miR-206 level was decreased in the moderate stenosis group and the severe stenosis group(P＜0.05).Compared with moderate stenosis group,EVD,PSV and serum miR-222-3p levels were significantly in-creased,and miR-206 level was decreased in severe stenosis group(P＜0.05).EVD,PSV,serum miR-222-3p level were risk factors for carotid artery stenosis in patients with cerebral infarction,and serum miR-206 level was a protective factor for carotid artery stenosis in patients with cerebral infarction(P＜0.05).The area un-der the curve of EVD,PSV combined with serum miR-222-3p and miR-206 in diagnosing the degree of carotid artery stenosis in patients with cerebral infarction was 0.991,and the sensitivity and specificity were 98.36％and 93.10％,respectively.It was better than that of EVD,PSV,serum miR-222-3p,and miR-206 alone(Zcombination-EVD=2.927,P=0.003,Z combination-PSV=2.834,P=0.005,Zcombination-miR-222-3p=3.300,P=0.001,Zcombination-miR-206=3.730,P＜0.001).Conclusion Cervical vascular color Doppler ultrasound combined with ser-um miR-222-3p and miR-206 has application value in the diagnosis of carotid artery stenosis in patients with cerebral infarction.

Objective:To investigate the association of polymorphisms at the matrix metalloproteinase(MMP)-3-1612 position of the promotor region with the inflammatory response and oxidative stress in elderly patients with cerebral ischemic stroke.Methods:In this retrospective study, 129 elderly patients with cerebral infarction diagnosed and treated in our hospital between March 2019 and March 2021 were enrolled as the study group, and 110 healthy subjects were selected as the control group.Polymorphisms of the MMP-3-1612 position in the promotor region, the inflammatory response and oxidative stress were examined using appropriate parameters and the associations between them were analyzed. Results:Compared with the control group, the proportions of patients with hypertension, diabetes, and smoking history in the study group were significantly higher( χ2=16.05, 17.19, 14.19, all P<0.05), and the levels of fasting blood glucose, low-density lipoprotein, and homocysteine were also significantly higher( t=6.22, 3.64, 2.69, all P<0.05).Meanwhile, compared with patients carrying the MMP-3-5A/6A or the MMP-3-6A/6A genotype, the levels of serum inflammatory markers such as high mobility group box-1 protein(HMGB1), fractalkine(FKN), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β)and interleukin-17(IL-17)in patients carrying the MMP-3 gene 5A/5A genotype were significantly higher(all P<0.05).In addition, the expression of serum oxidative stress-related molecules Kelch-like ECH-associated protein 1(Keap1), nuclear factor erythroid-2 related factor2(Nrf2), antioxidant response element(ARE), quinone oxidoreductase 1(NQO1), and heme oxygenase-1(HO-1)was also significantly increased(all P<0.05), but there was no difference in these markers between patients carrying the MMP-3-5A/6A genotype and patients carrying the MMP-3-6A/6A genotype( P>0.05).Patients carrying the 5A/5A genotype and the 6A/6A genotype exhibited only one 97 bp band and one 120 bp band, respectively, while the patients carrying the 5A/6A genotype exhibited two 97 bp bands and two 120 bp bands.There was no statistical difference in the number of patients carrying the 5A/6A genotype in the cerebral infarction group compared with the control group( P>0.05), and the number of patients carrying the 5A/5A genotype in the cerebral infarction group was higher than that in the control group(69% or 53.49% vs.35% or 31.82%, χ2=11.34, P<0.05).Polymorphisms of the MMP-3 gene had a positive correlation with the risk of stroke( r=0.25, P<0.05). MMP-3-1612 gene polymorphism( OR=7.21, 95% CI: 1.13-1.83, P=0.01), elevated blood glucose( OR=1.27, 95% CI: 1.18-2.06, P<0.001), high homocysteine( OR=1.05, 95% CI: 1.08-1.58, P<0.01), hypertension( OR=5.414, 95% CI: 1.140-4.46, P<0.01), elevated low-density lipoprotein( OR=4.03, 95% CI: 1.03-2.35, P=0.02), coronary heart disease( OR=1.17, 95% CI: 1.47-3.19, P<0.01)and diabetes( OR=8.52, 95% CI: 1.32-4.71, P<0.01)were risk factors for cerebral infarction. Conclusions:In elderly patients with cerebral infarction, polymorphisms of the MMP-3-1612 position in the promotor region is closely related to the risk of cerebral ischemic stroke, the inflammatory response and oxidative stress.MMP-3 gene polymorphisms are risk factors for stroke.

OBJECTIVE We used Al lergic Rhinitis Control Test (ARCT) questionnaire to evaluate the control level of allergic rhinitis, in order to get the data of prevalence rate, epidemiological characters and risk factors of uncontrolled allergic rhinitis. METHODS Patients with AR were recruited from our department and the treatment based on the Allergic Rhinitis and Its Impact on Asthma(ARIA) guidelines. Telephone interview will be taken after 2 weeks in these patients, compared to symptoms, impact on quality of life, ARCT value before and after treatment. RESULTS Among 134 patients enrolled, moderate/severe AR account for 95％. After 2 weeks of treatment, both symptom and quality of life were marked improvement(P <0.001). Patients with uncontrolled AR(26.1％) at day 15 more frequently presented higher height and weight(P <0.001), history of ear, nose, and throat(ENT) infection or antibiotics intake for respiratory infection in the last 12 months (40.4％ versus 62.9％, P =0.022), smoking (4.0％ versus 17.1％, P =0.02), and smell disturbance (10.1％ versus 25.7％, P =0.044). CONCLUSION Most of AR patients have remarkable improvements in symptom and quality of life after treatment, but 26.1％ of patients still remain uncontrolled. Smoking and nose infection are risk factors of uncontrolled AR.