Objective To investigate the effects of DNA methylation on the expression of lympho-cyte activation gene 3 (lag3) in different human T cell lines.Methods A quantitative PCR and a flow cy-tometry analysis were performed to measure the expression of lag3 gene in various T cell lines at mRNA and protein levels.The distribution of CpG sites within the promoter and body of lag3 gene was detected to locate the potential regulatory region(s) (CpG island and CpG island shore).The levels of DNA methylation in each cell line were analyzed.The T cell lines were demethylated with 5-Aza-2′-deoxycytidine (5-Aza-2′-dc) for further investigation on the changes of lag3 gene expression and DNA methylation.Results Jurkat E6-1 cells showed the highest expression level of lag3 gene as compared with J.CaM1.6 and CEM cells.Hyperm-ethylated CpG islands were detected in cells of each cell line.The methylation levels of CpG island shore in J.CaM1.6 and CEM cells were higher than that in Jurkat E6-1 cells.Treatment of J.CaM1.6 and CEM cells with 5-Aza-2′-dc significantly promoted the expression of lag3 gene at mRNA and protein levels as well as the demethylation of CpG island shore.No significant differences with the expression of lag3 gene and the methylation of CpG island were observed in Jurkat E6-1 cells with or without 5-Aza-2′-dc stimulation.Con-clusion Methylation and demethylation of CpG island shore played important roles in regulating the tran-scription of lag3 gene.

Objective To study in primary nephrotic syndrome clinical effect in the treatment of statins combined with glucocorticoid for syndrome disease.Methods From May 2013 to February 2017 treatment of 90 cases of primary nephrotic syndrome patients as the research object, the patients were divided into control group and the application of the experimental group randomly, the control group received glucocorticoid therapy, the experimental group patients in the control group based on the addition of statin therapy, compared two groups of patients after treatment of triglyceride (TG), serum total cholesterol (TC), serum CRP, serum creatinine, two D dimer And 24 h urinary protein and clinical curative effect.Results After the treatment, the patients in the experimental group TG, TC, blood CRP, serum creatinine, two D dimer and 24 h urinary protein equivalents were significantly better than the control group (P<0.05);the clinical treatment of patients in the experimental group the total efficiency of 95.56% was significantly higher than that of the control group (73.33%), the difference was statistically significant(P<0.05).Conclusion Statins combined with glucocorticoids in the treatment of primary nephrotic syndrome can effectively improve serum albumin, reduce urinary protein content, promote the recovery of renal function, clinical curative effect, worth in clinical treatment.Line promotion application.

Objective To investigate the expression and clinical significance of mammalian sterile 20-like kinase 1 (MST1) in cervical cancer. Methods Immunohistochemical method was applied to detect the expression level of MST1 protein in specimens of cervical cancer tissues (n=139) and pericarcinomatous tissues (n=20, with≥4 cm distance from the primary tumor's edge). Western blot assay and qPCR were used to detect the protein and mRNA transcription expression levels of MST1 in 20 pairs of cervical cancer tissues and pericarcinomatous tissues, respectively. The correlation between MST1 expression, clinic pathological features and the prognosis were analyzed. Results MST1 was mainly expressed in cytoplasm. The positive expression rate of MST1 was significantly lower in cervical cancer tissues (27%, 38/139) than that in pericarcinomatous tissues (80%, 16/20,χ2=21.62, P<0.01). The expressions levels of MST1 protein and mRNA were both lower in the cervical cancer tissues (P<0.01). In cervical cancer, the positive expression rate of MST1 inⅠb+Ⅱa stage was higher than that ofⅡb+Ⅳstage (P<0.05), the positive expression rate of MST1 in lymph node metastasis was lower than that of without lymph node metastasis (P < 0.05). Values of age, tumor size, histological type and differentiation degree showed no significant difference to positive expression rate of MST1. Moreover, the negative expression of MST1 displayed a significantly poorer overall survival time than that of positive expression of MST1 (Log-rank χ2=28.35, P < 0.01). Conclusion MST1 shows a lower expression in cervical cancer, which may be a new target for clinical treatment and prognosis of cervical cancer.

Objective To understand the etiology of acute hepatitis B (AHB) in adults and investigate the mechanisms of hepatic injury and viral clearance in AHB. Methods One hundred and twenty adult AHB patients were enrolled. Epidemiological and clinical data were collected from the case history records or face-to-face inquiry, and serum samples were collected during hospitalization and follow-up. To observe dynamic patterns of AHB etiology, the markers of hepatitis B virus (HBV) were detected by enzyme-linked immunosorbent assay (ELISA); the level of HBV DNA and HBV genotype were determined by real-time polymerase chain reaction (PCR). Enumeration data were analyzed by non-parametric rank sum test. Comparison between groups was done by t test and that between rates of samples was done by Pearson χ2 test. Results Serum HBV DNA was positive in 48.33% of patients at the time of diagnosis with mean level of 9.84×04 copy/mL, and became undetectable after 12.5 days on average. The median levels of serum alanine aminotransferase (ALT) were 1600 U/L and 1490 U/L in HBV DNA positive and negative groups, respectively (z=-0. 678, P=0. 498). However, the mean levels of serum ALT were (2058±123) U/L and (1393±139) U/L in groups of HBV DNA<1×104 copy/mL and>1×104 copy/mL, respectively, which was significantly different (t=-2.17, P=0. 049). Genotype B accounted for 52.5%, genotype C 42.5 and genotype B and C mixed type 5.0% in 58 patients with HBV DNA positive. Eight patterns of serum HBV markers were presented at first visiting. HBsAg(+), HBeAg(+), anti-HBc(+), anti-HBc IgM(+) and HBsAg(+), anti-HBe(+), anti-HBc(+), anti-HBc IgM(+) were the most common patterns, which accounted for 38.3% and 30.0%, respectively. The dynamic patterns of serum HBV markers of 28 AFIB patients were prospectively followed up. The rate of serum FIBsAg loss was 100. 0% and the median time of negative-conversion was 3 weeks. The cumulative positive rate of anti-HBs was 85.7% after 52 weeks of follow-up. The rate of serum HBeAg loss was 100.0%. HBeAg was negative in 53.6% of patients at first visiting and the rest of patients achieved negative within 4 weeks after onset. The positive rate of anti-HBe was 82.1% during 52 weeks of follow-up. Total anti-HBc (including IgG and IgM) was keeping positive in all patients within 52 weeks, and the negative rate of anti-HBc IgM was 39. 3% after followed up for 52 weeks. Conclusions Rapid HBV clearance andserum HBV marker conversion are significantly different between AHB and chronic hepatitis B.

OBJECTIVE:To investigate the clinical efficacy and safety of Recombinant human interferon α2b(rhIFN α2b) vaginal effervescent capsules combined with radiofrequency ablation in the treatment of cervical erosion with human papilloma virus (HPV)subclinical infection(SPI). METHODS:A total of 207 cervical erosion patients with SPI were selected from gynecology outpatient department of our hospital during Jul. 2014-Aug. 2015 and then divided into group A,B,C according to random number table,with 69 cases in each group. Group A was given rhIFN α2b vaginal effervescent capsules 800 thousand IU,via posterior for-nix,qd,3 days after the end of menstruation,10 days as a treatment course,for 3 courses. Group B received radiofrequency abla-tion. Group C was given constant dose of rhIFN α2b vaginal effervescent capsules combined with radiofrequency ablation. The clini-cal efficacy of 3 groups,the rate of wound healing,the rate of associated symptoms disappearance 2 weeks after surgery and the in-cidence of complications in group B and C were evaluated. The occurrence of ADR was recorded. RESULTS:The response rates of group B,C were 94.20% and 98.55%,which were significantly higher than 62.32% of group A. The response rates of SPI in group C was 92.75%,which was significantly higher than 63.77% of group B,with statistical significance(P<0.05). The wound healing rates of group B 4,6,8 weeks after surgery were 10.14%,43.48%,97.10%;and those of group C were 52.17%,92.75%, 100.00%.The wound healing rates of group C 4,6 weeks after surgery were significantly higher than those of group B,with statisti-cal significance(P<0.05).The disappearance incidence of symptom as vaginal bleeding and drainage in group C 2 weeks after sur-gery was 81.16%,which was significantly higher than 43.48% of group B,with statistical significance(P<0.05).The incidences of complications were 11.60% in group B and 4.35% in group C,without statistical significance(P>0.05). No obvious ADR was found in 2 groups. CONCLUSIONS:rhIFN α2b vaginal effervescent capsules combined with radiofrequency ablation can effective-ly improve the efficacy of cervical erosion with SPI,shorten the wound healing time with good safety.

Objective To discuss the effect of MST1 (mammalian sterile 20-like kinase 1,MST1) on the prolifera-tion,migration and invasion of SiHa cervical cancer .Methods Western blot was used to detect the expression of MST1 in cervical epithelial cells H8 and cervical cancer cells SiHa;PJ3H-HA-MST1 was constructed and transfect-ed it to SiHa cells by Lipofectamine TM3000;MST1, Ki-67 and MMP9 protein expression were evaluated by Western blot;While the proliferation ,migration and invasion of SiHa cell were assessed by MTS ,scratch adhesion test and Transwell assay respectively .Results Compared SiHa cells with H 8 cells,MST1 expression in SiHa cells was sig-nificantly lower than that in H8 cells.The plasmid was successfully transfected into SiHa cells , MST1 expression was significantly higher , while the expression of Ki-67 and MMP9 was lower .The proliferation , migration and inva-sion ability were all significantly suppressed .Conclusions Overexpression of MST1 can inhibit the proliferation , migration and invasion of cervical cancer cell line SiHa .

Objective:To investigate the relation between rs2298771 genotype in voltage-gated sodium channels 1A ( SCN1A) polymorphism and antiepileptic drug (AED) response in children with epilepsy. Methods:Sixty-two children with epilepsy admitted to Department of Neurology, Zhangjiakou First Hospital from June 2022 to December 2023 were divided into AED response group and AED resistance group ( n=31) according to their response to AED. In addition, 31 children with pharyngitis or mild gastroenteritis admitted to Department of Pediatrics at the same period were selected as control group. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to analyze the rs2298771 genotype in SCN1A polymorphism, and differences in rs2298771 genotype and allele in SCN1A polymorphism were compared among the 3 groups. Relation between rs2298771 genotype in SCN1A polymorphism and AED response was analyzed. Multivariate Logistic regression was used to analyze the influencing factors for AED response in children with epilepsy. Results:(1) Significant differences in type of first seizure and AEDs were noted between AED response group and AED resistance group ( P<0.05); compared with the AED resistance group, the AED response group had significantly lower seizure frequency, significantly longer duration after last seizure, and statistically higher proportions of children with normal EEG or with one kind of AED ( P?0.05). (2) Compared with the control group and AED response group, the AED resistance group had significantly higher rs2298771 GC genotype and G allele, and statistically lower rs2298771 AA genotype and A allele in SCN1A polymorphism ( P?0.05). (3) In the AED response group, rs2298771 AA and AG genotype in SCN1A polymorphism were positively correlated with levetiracetam ( P?0.05); in AED resistance group, rs2298771 AG genotype in SCN1A polymorphism was positively correlated with topiramate and valproic acid ( P<0.05). (4) Multivariate Logistic regression analysis showed that duration after last seizure ( OR=3.249, 95% CI=1.097-9.621, P=0.033), rs2298771 genotype in SCN1A polymorphism ( OR=9.660, 95% CI=4.680-19.970, P=0.011) and seizure frequency ( OR=0.160, 95% CI=0.032-0.804, P=0.026) were independent influencing factors for AED response in children with epilepsy. Conclusion:Epilepsy children with shorter duration after last seizure, rs2298771 GG genotype in SCN1A polymorphism, and high seizure frequency are susceptible to AED resistance; especially, AG genotype is correlated with topiramate and valproic acid.

Objective:To investigate the characteristics, causes and preventive strategies of intraoperative and postoperative complications of Da Vince robotic radical resection for lung cancer.Methods:From January 2018 to June 2020, 306 patients with lung cancer who underwent robotic radical resection of lung cancer in our department were reviewed, the perioperative data were statistically analyzed. There were 154 males and 152 females, aged(58.5±10.3) years old, 238 lobectomy cases and 68 segmental lobectomy cases.Results:There were no perioperative death, no conversion to thoracotomy, and no intraoperative vascular injury. Intraoperative blood loss was(41.5±37.4)ml, 302 cases(98.7%) underwent R0 resection, 54 cases(17.6%) of intraoperative bleeding from troca mouth of robot operating arm. 32 cases(10.5%) of postoperative complications, including 3 cases(1.0%) of chylothorax, 1 case(0.3%) of pleural effusion, 28 cases( 9.2%) of alveolar fistula over 7 days with 5 cases of large area subcutaneous emphysema.Conclusion:The most common intraoperative complication in robotic lung cancer radical operation is troca bleeding, and the most common postoperative complication is alveolar fistula. Robot surgery is safe, and targeted preventive measures can reduce the incidence of complications.

Objective:To report embryonic testicular regression syndrome(ETRS) caused by DHX37 heterozygous variant for the first time in China and summarize the clinical manifestations of ETRS as to improve the understanding of doctors for this disease.Methods:The clinical data and whole exome sequencing results of five cases of ETRS from Shenzhen Children′s Hospital were collected. The reported cases of DHX37 heterozygous variant were reviewed.Results:Five patients with ETRS visited the doctors at the age of 2 months to 5 years and 5 months. Three patients raised as males came to hospital due to virilition and 2 female patients visited a doctor due to clitoral hypertrophy. No uterus was detected by ultrasound in all patients. The gonadal pathologies from 4 cases displayed no testicular tissue or gonadal dysgenesis, complicated with gonadoblastoma in one case. The genetic testing revealed that the heterozygous variant(c.923G>A, p. R308Q) in DHX37 was found in 2 cases, without variant in other 3 cases. According to the review, ETRS and 46, XY gonadal dysgenesis due to DHX37 herozygous variant was firstly reported in 2019. A total of 40 cases, including 21 cases of ETRS, presented with the virilition or female phenotype, with the disappearance of testicular tissue as the main pathologies. There is no report in China.Conclusion:The article summarized the clinical manifestations and whole exome sequencing results of 5 patients with ETRS, among which two cases were caused by DHX37 variants and one was complicated with gonadoblastoma.

Objective To establish and evaluate chemotherapeutic phlebitis model rats induced by vinorelbine via the dorsalis pedis vein.Methods Rats were divided randomly into control and 4 different concentration of vinorelbine-induced model groups.Control rats were injected with 0.1 mL normal saline via the dorsalis pedis vein of the hind limb,while other rats were injected with different concentrations of vinorelbine(2,3,4,5 mg/mL),as above.General observations were performed and the hind limb volume was measured daily for 7 consecutive days to calculate the swelling rate.The rats were then killed and histological changes in the dorsalis pedis vein were observed by hematoxylin and eosin staining.Microstructural changes on the surface of the vascular endometrium were observed by scanning electron microscopy.Results Injection of 2,3,4,5 mg/mL vinorelbine via the dorsalis pedis vein significantly induced hind limb swelling in a concentration-dependent manner,peaking on day 3 in each group.The phlebitis rates on day 7 were 50%in the 2 mg/mL group and 83.3%in the 3 mg/mL group.Phlebitis was also induced in the 4 mg/mL and 5 mg/mL groups,including grade Ⅲ in 66.6%and grade Ⅳ in 83.3%.Histopathology showed inflammatory cell infiltration,wall thickening,lumen stenosis,and thrombosis in the tissues surrounding the veins.Scanning electron microscopy showed destruction of tight junctions of venous endothelial cells,and a rough surface of the vascular lining,resultsing in blood cell adhesion.Conclusions Injection of 0.1 mL of 3～5 mg/mL vinorelbine via the dorsalis pedis vein could induce red,swollen,and cord-like veins,as well as infiltration of inflammatory cells around the vein,thickened vein walls,lumen stenosis,and thrombosis.In addition,the surface of the venous intima was rough and adhered to numerous blood cells.All these features are consistent with those of clinical chemotherapeutic phlebitis in terms of the symptoms and pathological structure.