[ABSTRACT]OBJECTIVETo investigate the cytomegalovirus infection in neonates, characteristics of gap junction protein Connexin26 gene mutation and the hearing follow-up results, and to analyze their correlations. METHODS60 CMV-DNA positive and 40 CMV-DNA negative neonatal newborn from The Second Affiliated Hospital of Wenzhou Medical University and The first people's Hospital of Yongkang were screened, the blood biochemistry was analyzed, and the umbilical cord blood was reserved to detect the Connexin26 gene expression of mRNA with RT-PCR.PCR results was sequenced to track the newborn hearing, and analyze the correlations between neonatal cytomegalovirus types, the mutation of Connexin26 gene and hearing test results.RESULTS 26 cases from 60 CMV-DNA positive newborns were found with blood biochemical abnormalities. In all of the newborns, a total of 41 cases had 235delC mutation, 11 cases in the mutations for the development of hearing impairment. The results of correlation analysis showed that there were correlations between cytomegalovirus infection, gene mutation and hearing impairment.CONCLUSION Cytomegalovirus infection in neonates can lead to mutations in the Connexin26 gene, and may further lead to hearing impairment, and the probability of the mutation of Connexin26 gene and sensorineural hearing loss were higher in symptomatic cytomegalovirus infection neonates.

Objective: To investigate the efficacy of treating craniocerebral trauma-related coma with acupuncture plus hyperbaric oxygen. Methods: Coma patients were randomly allocated into observation and control groups. Routine surgery or non-surgical treatment was given according to the condition of injury. The observation group received acupuncture plus hyperbaric oxygen as well as the treatment for the control group. The score was counted by the Glasgow coma scale before and after the treatment. A chi-square test was used. Results:There was a very significant difference in curative effect between the observation and control groups (P＜0.01). Conclusion: Treatment with acupuncture plus hyperbaric oxygen can promote the patients' early revival.

Objective To investigate the effects of arsenic trioxide(ATO) on anti-dsDNA antibody and lymphocyte subsets proliferation in splenic cells of MRL/lpr mice.Methods MRL/lpr mice were divided into three groups: ATO group(0.4 mg/kg?d,ip),sodium chloride(NS) group(0.25 mL,ip),and cyclophosphamide(CTX) group(50 mg/kg?qw,ip).The control group consisted of 12 syngeneic normal C57/BL mice,which were sub-divided into ATO group(0.4 mg/kg?d,ip) and NS group(0.25 mL,ip).After two-month treatment,all mice were killed and their bodies and spleens were weighed.Anti-dsDNA antibody in serum were detected by ELISA.The percentage of different cell subsets was checked through flow cytometry.Results(1)The spleen index、serumlevel of anti-dsDNA antibody and the percentages of CD19+、CD3+、CD3+CD4+ lymphocytes in ATO group were much lower than those in NS group of MRL/lpr mice(P0.05). Conclusion Arsenic trioxide can inhibit the activation of T and B lymphocytes,increase the percentage of NK cell and reduce the serum level of auto-antibody.And it was of no effect in the normal mice.

Objective:To explore the effect of fecal microbiota transplantation (FMT) on the formation of renal calcium oxalate crystals in SD rats induced by oxalate mixed diet.Methods:Six male guinea pigs were fed with standard guinea pig chow for 1 month and then given a 5% oxalate diet for 14 d. The guinea pigs on the standard chow were labeled as the standard chow guinea pig (GSC group) and those on the high oxalate diet for 14 d were labeled as the guinea pig group on the high oxalate diet (GOD group). The feces of guinea pigs in the GSC and GOD groups were collected using metabolic cages. Twenty-four male SD rats were randomly divided into standard chow (SC) group, oxalate diet(OD)+ phosphate buffered saline gavage group (OD+ PBS group), OD+ FMT group and SC+ FMT group. Among them, the SC group and SC+ FMT group were fed with standard chow. The OD+ PBS group and OD+ FMT group were fed with 5% oxalate content chow. The OD+ FMT and SC+ FMT groups were given GOD group guinea pig fecal filtrate gavage for 7 days. The 24 h urine and feces of rats in each group were collected, and the intestinal microbiota of rats and guinea pigs were detected by 16sRNA detection. The urinary oxalate excretion was detected by high performance liquid chromatography. The rats and kidneys were weighed and the renal index was calculated. HE staining was used to observe the histological morphological changes of rat kidney tissue, the calcium oxalate crystal deposition in renal tissues was detected by Pizzolato staining.Results:The relative abundance of bacteria from a total of 11 families, including Muribaculaceae family and Bifidobacteriaceae family, was significantly increased in the intestinal tract of guinea pigs (GOD) from the high oxalate diet group compared to guinea pigs (GSC) from the standard chow group. The microbial diversity of the intestinal microbiota of the rats in the OD+ PBS group was reduced compared to the SC group, and the microbial diversity of the intestinal microbiota of the rats in the OD+ FMT group was restored compared to the OD+ PBS group. When given a standard chow, the intestinal microbiota of rats receiving FMT deviated from that of normal rats and was more similar to that of guinea pigs fed a high oxalate diet. In the OD+ FMT group, bacteria from a total of 18 families, including Muribaculaceae family, Erysipelotrichaceae family and Bifidobacteriaceae family, were significantly enriched, and FMT activated the intestinal microbial network represented by bacteria from Muribaculaceae family. The renal index of rats in the OD+ PBS group was significantly increased compared to the SC group (7.63±0.67 vs. 6.12±0.53, P<0.05), whereas the renal index of rats in the OD+ FMT group was significantly decreased in comparison to the OD+ PBS group (6.53±0.64 vs. 7.63±0.67, P<0.05). Urinary oxalate excretion of rats in the SC group, the OD+ PBS group, and the OD+ FMT group were (0.61±0.05), (0.89±0.04) and (0.72±0.04) μmol/ml, respectively. In the rats of the SC group no calcium oxalate crystals were seen in the kidney (0 score) and more calcium oxalate crystals were detected in the OD+ PBS group (4.83±0.41 score). The OD+ FMT group showed significantly lower calcium oxalate crystallization scores (3.17 ± 0.75 score, P<0.01) compared to the OD+ PBS group. Conclusions:FMT activated the microbial network represented by bacteria from the family Muribaculaceae in the rat intestine, significantly reduced urinary oxalate excretion and renal calcium oxalate crystal deposition in rats on a high oxalate diet.

Objective:To investigate the long-term outcome and prognostic indicators of diffuse pro-liferative lupus nephritis (DPLN).Methods:The primary endpoint of long-term follow-up and factors pos- sibly influencing the outcome were analyzed retrospectively in DPLN patients admitted to the First Affiliated Hospital of Wenzhou Medical University between Jan 1, 2000 and Dec 31, 2014. Patients were classified into three groups according to the evaluated glomerular filtration rate(eGFR) on the first day of admission: eGFR≥60 ml·min -1·1.73 m -2 (regular illness group); 15 ml·min -1·1.73 m -2≤eGFR<60 ml·min -1·1.73 m -2 (serious illness group); eGFR<15 ml·min -1·1.73 m -2 or dialysis (critical illness group). Clinical, histological, and outcome differences among the three groups were evaluated and compared using one-way analysis of variance (ANOVA) , χ2 test, Kaplan-Meier survival curve and Cox reggression analysis. Results:167 DPLN patients were studied [155 women; mean age (30±10) years; mean follow-up of (61±45) months]. Renal and patient survival of all patients was 86% at 5 years and 79% at 10 years. Kaplan-Meier analysis showed the renal and patient survival rate at 10 years in the regular illness group, serious illness group and critical illness group was 91%, 70% and 8%, respectively ( χ2=121.93, P<0.01, overall); regular illness group vs serious illness group ( χ2=4.05, P<0.05); regular illness group vs critical illness group ( χ2=97.05, P<0.01); serious illness group vs critical illness group ( χ2=52.28, P<0.01). Multivariable Cox regression analysis found that haematoglobin (Hb)<80 g/L [ HR=2.7, 95% CI(1.2, 6.3), P=0.019], eGFR<60 ml·min -1·1.73 m -2 [ HR=4.1, 95% CI(2.0, 8.2), P<0.01] and large crescents ≥30%[ HR=1.8, 95% CI (1.1, 2.9), P=0.021], were risk factors for the long-term outcome. Conclusion:DPLN patients with normal or slightly decreased renal function have a better long-term prognosis. Moderate to severe impairment of renal function, anemia and large crescents are associated with poor outcome.

Objective To explore the effects of fecal microbiota transplantation(FMT)on oxalate metabolism and renal protection in rats fed a high oxalate diet.Methods Twenty-four male SD rats were randomly divided into four groups:SC,SC+FMT,OD+PBS and OD+FMT.The SC group was set as the control group and was fed standard rat chow.The OD+PBS group and OD+FMT group were fed a diet containing 5%oxalate.Starting from day 14,the OD+PBS group,OD+FMT group and SC+FMT group received intragastric administration of PBS solution or filtered faecal microbiota solution from guinea pigs for 7 consecutive days.The 24-hour urine,feces,and venous serum of the rats were collected from the rats of all groups to determine the gut microbiota and biochemical markers.Real-time quantitative PCR and immunohistochemistry were conducted on the rat kidneys to detect the expression of renin,ACE,and OPN.Results The fecal microbiota transplantation altered the gut microbiota of rats.The gut microbiota of the SC+FMT group deviated from that of the SC group and showed increased similarity to that of the guinea pigs.Compared to the OD+PBS group,the OD+FMT group exhibited significant reductions in the urinary oxalate,urinary urea,uric acid,urinary creatinine,serum urea nitrogen/creati-nine,and serum uric acid.Furthermore,after FMT treatment,the OD+FMT group exhibited reduced upregulation of renin mRNA expression and restored downregulation of OPN mRNA expression compared to the OD+PBS group;similar results were obtained from immunohistochemistry.Conclusion Fecal microbiome trans-plantation activated the microbial network in the rat gut,particularly the oxalate-degrading bacteria represented by Muribaculaceae.The kidney injury induced by high oxalate was partially restored by the microbiota network's degradation of oxalate,indicating the protective effect of fecal microbiota transplantation on the rat kidneys.

Objective To explore the safety and efficacy of neoadjuvant chemotherapy(NAC)combined with immunotherapy before radical cystectomy plus pelvic lymph nodes dissection(RC-PLND)for muscle-invasive bladder cancer(MIBC).Methods The clinical data of 101 patients with MIBC who underwent neoadjuvant therapy followed by RC-PLND in the Department of Urology,the First Affiliated Hospital of Xi'an Jiaotong University during Jan.2019 and Dec.2023 were retrospectively analyzed,including 71 patients(70.3％)who received NAC(NAC group)and 30(29.7％)who received NAC combined with immunotherapy(NAC combine immunotherapy group).The clinical and pathological data and adverse events during neoadjuvant therapy were compared.Logistic regression analysis was used to explore the independent predictors of pathological complete response(pCR)and pathological partial response(pPR).Results There were no significant differences in the baseline data between the two groups(P＞0.05).However,the proportion of multiple tumors in patients receiving NAC before surgery was significantly higher than that in the NAC combined immunotherapy group(69.0％vs.46.7％,P=0.034).Compared with NAC group,NAC combined with immunotherapy group had significantly improved rate of pathological downstaging and pPR(60.6％vs.83.3％,P=0.026;45.1％vs.70.0％,P=0.022).Furthermore,the rate of pCR in patients undergoing NAC combined immunotherapy was higher than those undergoing NAC,but the difference was not significant(53.3％vs.33.8％,P=0.067).Logistic regression analysis revealed that clinical T-stage and tumor diameter were independent predictors of pCR and pPR(P＜0.05).In addition,the most common adverse events during neoadjuvant therapy were anemia,decreased white blood cells,nausea,and vomiting,but most of them were grade 1-2 and could be relieved through symptomatic treatment.Conclusion NAC combined with immunotherapy is safe and effective,which can improve the rate of pathological downstaging,pPR and pCR,without increasing the incidence of adverse reactions.

Objective:To explore the effects of varicella-zoster virus (VZV) on the glycosylation characteristics of cellular prion protein (PrP C) in human Schwann cells (hSC) and the regulation of methylcobalamin (MeB 12). Methods:The hSC were inoculated with VZV at 1.0 multiplicity of infection for 48 hours, then 250 μg/ml of MeB 12 were added and cultured for 48 hours. PrP C from the supernatant and sediment were coated with anti-PrPC antibody (3F4) respectively and subjected to screening for glycans by sandwich lectin-ELISA. Meanwhile, the superoxide dismutase (SOD) and malondialdehyde (MDA) from the supernatant were detected by diagnostic reagent kit. Results:The ratio of PrP C glycans in the supernatant to sediment of VZV-infected cells was found to be significantly different compared with those in the VZV-non-infected cells. The overall glycans ratios of the supernatant to the sediment was 1∶2.6 in the uninfected cells, while the ratio was 1∶1.5 in the VZV-infected cells (F=24.18, P<0.001, LSD-t=8.27, P<0.001), suggesting that stability of PrP C decreased after VZV infection, and correspondingly the activity of SOD (4.43±2.05 U/mg) was significantly reduced in the VZV-infected hSCs compared with those(14.23±1.27 U/mg) in the uninfected cells (F=18.19, P=0.001, LSD-t=6.54, P<0.001), the level of MDA (11.17±1.89 nmol/mg) was significantly elevated in the VZV-infected hSCs compared with those (3.73±0.35nmol/mg) in the uninfected cells (F=30.70, P<0.001, LSD-t=8.25, P<0.001). When the VZV-infected cells were added with 250 μg/ml MeB 12, glycans in the sediment of infected cells significantly increased compared with those in the VZV-infected cells without MeB 12, the overall glycans ratio of the supernatant to the sediment was 1∶2.4, suggesting that MeB 12 improved the stability of PrP C. Moreover, SOD activity (11.07±2.07 U/mg) was significantly increased (LSD-t=4.42, P=0.002), MDA level (5.23±0.96 nmol/mg) was significantly decreased (LSD-t=6.58, p<0.001) in the VZV-infected cells added with MeB 12 compared with those in the VZV-infected cells without MeB 12. Conclusions:The glycosylation characteristics of PrP C in hSC could be changed by VZV, while MeB 12 could regulate the glycosylation characteristics to improve the stability of PrP C, thereby increase the antioxidant capacity of hSC.

Objective: To investigate the clinico-pathological characteristics, outcomes and their predictors in malignant hypertension related kidney injury with and without primary glomerular diseases. Methods: Patients with clinical diagnosis of malignant hypertension, biopsy-proven kidney injury caused by malignant hypertension and complete clinical data from January 2010 to December 2018 were retrospectively analyzed. According to clinical and renal pathology, patients were divided into malignant hypertension related kidney injury without primary nephropathy group and with primary nephropathy group. Clinico-pathological characteristics and outcomes were evaluated and compared between malignant hypertension related kidney injury with and without primary glomerular diseases. Results: Totally 31 biopsy-proven kidney injury patients were analyzed. Among them, there were 18 cases with primary glomerular diseases and 13 cases without primary glomerular diseases, with age of (32.5±6.5) years old and (34.7±8.1) years old, respectively. There were 12 males in both group. The proportion of primary IgA nephropathy was higher (16/18) in the group of malignant hypertension related kidney injury with primary glomerular diseases. Malignant hypertension with primary glomerular diseases patients had lower plasma albunin level [(32.7±6.4) g/L vs (38.5±7.3) g/L, P=0.027], higher 24-hour proteinuria level [(4.03±2.71) g vs (1.45±0.98) g, P=0.002] and higher incidence rates of dysmorphic hematuria (14/18 vs 0, P=0.001) than those without primary glomerular diseases patients. Glomerular sclerosis, mesangial proliferation, tubular atrophy and interstitial fibrosis were more severe in malignant hypertension with primary glomerular diseases patients (all P<0.05), but the ischemic wrinkling of glomerular capillary was more severe in malignant hypertension without primary glomerular diseases (P<0.01). There were no differences of acute or chronic malignant hypertensive injury in small artery and in afferent arterioles between the two groups. Cox regression analysis showed that loss of brush-border with flattening of tubular epithelium was the predictor for renal partial recovery (HR=5.956, 95% CI 1.198-29.614, P=0.029). Kaplan-Meier analysis showed that malignant hypertension patients with primary glomerular diseases had shorter renal survival time than those without primary glomerular diseases [(24.1±9.3) months vs (56.6±12.4) months], and accumulative renal survival rate of malignant hypertension patients with primary glomerular diseases was lower than that without primary glomerular diseases (11.6% vs 53.3%, Log-rank χ²=5.022, P=0.025). Multivariate Cox regression analysis showed that severe tubular atrophy and interstitial fibrosis were independent risk factors for end-stage renal disease in malignant hypertension patients (HR=5.870, 95% CI 1.372-25.112, P=0.017). Conclusions Malignant hypertension with primary glomerular diseases patients have more severe clinico-pathological renal impairment and poorer prognosis of long-term renal survival than those without primary glomerular diseases. Acute renal tubular injury (loss of brush-border with flattening of tubular epithelium) is the only predictor of renal function improvement in patients with malignant hypertension and renal impairment within one year. Tubular atrophy/interstitial fibrosis is a risk factor for end-stage renal disease in patients with malignant hypertension. Renal biopsy is an indispensable tool for predicting short-term and long-term renal outcomes.