Objective To explore whether hypertonic saline would partake in regulating Notch signaling in microglia in experimentally induced cerebral ischemic rats.Methods Male SD rats were randomly divided into sham group, cerebral ischemia group, normal saline group ( NS group ) , 10%hypertonic saline group (10%HS group) , the model of cerebral ischemia were established in all rats except the sham group by using middle cerebral artery occlusion ( MCAO) .After 2 hours of MCAO, the rats were through reperfusion for 24 h.In addition, rats in the normal saline group and 10% HS group were respectively treated with a continuous intravenous injection of normal saline (0.3 mL/h) and 10%HS (0.3 mL/h) by tail vein for 24 h.Immunofluorescence methods, RT-PCR and Western blot were used to detect the expression of Notch1 and intracellular Notch receptor domain ( NICD) .All data was analyzed by one-way analysis of variance ( ANOVA) , The intergroup comparisons were analyzed by the least-significant-difference (LSD) tests.Differences were considered statistically significant if P<0.05.Results Immunofluorescence showed that the expression of Notch1 and NICD were significantly increased in the microglia around peri-ischemia area in cerebral ischemia group and normal saline group compared to sham group;the expression of Notch1 and NICD in the microglia around peri-ischemia area were significantly reduced in 10% HS group compared to ischemia group and NS group.RT-PCR showed that the mRNA expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 1.000 ± 0.076; ischemia group: 2.203 ±0.283; NS group: 1.616 ±0.185; P <0.01 ); however, it was significantly reduced in 10% HS group compared to ischemia group and NS group ( ischemia group:2.203 ±0.283; NS group: 1.616 ±0.185; 10%HS group: 1.202 ±0.177; P <0.05 ) .Western blot showed that the protein expression of Notch1 was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.290 ±0.079; ischemia group: 0.750 ±0.029; NS group:0.765 ±0.182;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.750 ±0.029; NS group:0.765 ±0.182;10%HS group:0.390 ±0.195;P<0.05 ) .The protein expression of NICD was significantly increased in ischemia group and NS group compared to sham group ( sham group: 0.401 ±0.196; ischemia group: 0.906 ±0.359; NS group:0.847 ±0.153;P<0.01);but was significantly reduced in 10%HS group compared to ischemia group and NS group ( ischemia group:0.906 ±0.359; NS group:0.847 ±0.153;10%HS group:0.561 ±0.165;P<0.05 ) .Conclusion Our results suggest that HS markedly suppresses Notch signaling in microglia around the ischemia tissue area in experimental induced cerebral ischemic rats.

OBJECTIVETo study the chemical constituents from the 60% ethanol extract of Corydalis yanhusuo.

METHODThe chemical constituents were separated and purified by various chromatographic techniques, and their structures were elucidated by chemical evidence and spectroscopic analysis.

RESULTNine alkaloids were obtained and determined as 7-formyldidehydroglaucine (1), nantenine (2), (+)-O-methylbulbocapnine (3), d-corydaline (4), tetrahydrocoptisine (5), 8-oxocoptisine (6), palmatine (7), tetrahydropalmatine (8), and dehydrocorydaline (9), respectively.

CONCLUSIONCompound 1 was obtained from natural products for the first time, and its NMR data were firstly reported. Compound 3 was reported from Papaveraceae for the first time, and compound 6 was firstly reported from Corydalis yanhusuo.

Objective:To classify the symptom characteristics of young and middle-aged peritoneal dialysis patients and to explore the differences of demographic functional status and social function of patients with different symptom categories.Methods:A total of 179 peritoneal dialysis patients from 3 peritoneal dialysis centers in Shanghai were investigated from December 2019 to August 2020 by General Information Questionnaire, Peritoneal Dialysis Symptom Distress Scale. Latent class analysis was used to classify young and middle-aged peritoneal dialysis patients according to symptom characteristics. Multiple Logistic regression was used to explore the differences of demographic and disease characteristics of different categories of patients.Results:Peritoneal dialysis patients could be divided into three potential categories according to symptom characteristics ( P<0.05). According to the conditional probability of each category, they were named "low symptom group" (111 cases,62.0%), "high psychological-moderate physical symptom group" (22 cases, 12.3%), "high symptom group" (46 cases,25.7%). There were differences in working status ( OR=0.029, P<0.01), education level ( OR=152.799, P<0.01), duration ( OR=81.307, P<0.05), diabetic nephropathy ( OR=80.619, P<0.01) and CCI score ( OR=91.188, P<0.01) distribution among different potential categories of young and middle-aged peritoneal dialysis patients. Conclusions:There are three types of potential symptoms in young and middle-aged patients undergoing peritoneal dialysis. In clinical practice, medical staff should focus on the psychological status of young and middle-aged patients with low educational background and early stage of dialysis, and encourage them to return to work; at the same time, they should regularly evaluate the symptom burden of patients with diabetic nephropathy and high complication index peritoneal dialysis patients, in order to provide targeted intervention measures to prevent the progression of the disease.

Objective:To explore the effect of pediatric critical illness score (PCIS), pediatric risk of mortality Ⅲ score (PRISM Ⅲ), pediatric logistic organ dysfunction 2 (PELOD-2), pediatric sequential organ failure assessment (p-SOFA) score and Glasglow coma scale (GCS) in the prognosis evaluation of septic-associated encephalopathy (SAE).Methods:The data of children with SAE admitted to the Pediatric Intensive Care Unit (PICU), Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences from January 2010 to December 2020 were retrospectively analyzed. They were divided into the survival and death groups according to the clinical outcome on the 28th day after admission. The efficiency of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting death were evaluated by the area under the ROC curve (AUC). The Hosmer-Lemeshow goodness-of-fit test assessed the calibration of each scoring system.Results:Up to 28 d after admission, 72 of 82 children with SAE survived and 10 died, with a mortality rate of 12.20%. Compared with the survival group, the death group had significantly lower GCS [7 (3, 12) vs. 12 (8, 14)] and PCIS scores [76 (64, 82) vs. 82 (78, 88)], and significantly higher PRISM Ⅲ [14 (12, 17) vs. 7 (3, 12)], PELOD-2 [8 (5, 13) vs. 4 (2, 7)] and p-SOFA scores [11 (5, 12) vs. 6 (3, 9)] ( P<0.05). The AUCs of PCIS, PRISM Ⅲ, PELOD-2, p-SOFA and GCS scores for predicting SAE prognosis were 0.773 ( P=0.012, AUC>0.7), 0.832 ( P=0.02, AUC>0.7), 0.767 ( P=0.014, AUC>0.7), 0.688 ( P=0.084, AUC<0.7), and 0.692 ( P=0.077,AUC<0.7), respectively. Hosmer-Lemeshow goodness-of-fit test showed that PCIS ( χ2=5.329, P=0.722) predicted the mortality and the actual mortality in the best fitting effect, while PRISM Ⅲ ( χ2=12.877, P=0.177), PELOD-2 ( χ2=8.487, P=0.205), p-SOFA ( χ2=9.048, P=0.338) and GCS ( χ2=3.780, P=0.848) had poor fitting effect. Conclusions:The PCIS, PRISM Ⅲ and PELOD-2 scores have good predictive ability assessing the prognosis of children with SAE, while the PCIS score can more accurately evaluate the fitting effect of SAE prognosis prediction.

Objective:To investigate the prognostic factors of patients with pancreatic body and tail carcinoma.Methods:The clinical data of 64 patients with pancreatic body and tail carcinoma who underwent surgical resection or endoscopic ultrasound biopsy and were pathologically confirmed in the Second Affiliated Hospital of Jiaxing University from January 2013 to March 2020 were retrospectively analyzed. Age, gender, diabetes mellitus, serum CEA and CA19-9 levels at initial diagnosis, tumor site, maximum tumor diameter, TNM stage and treatment method were collected. Kaplan-Meier method was used to draw survival curve, and Log-rank test was used to analyze survival rate. Univariate and multivariate Cox proportional risk regression models were used for prognostic analysis.Results:Among the 64 patients, 24 patients were complicated with diabetes; serum CEA level was increased in 36 cases, and serum CA19-9 level was increased in 46 cases; 8 cases were in TNM stage ⅠA, 4 cases were in ⅠB stage, 4 cases were ⅡA stage, 4 cases were in ⅡB stage, 8 cases were in Ⅲ stage, and 36 cases were in Ⅳ stage. Symptomatic treatment was performed in 18 cases, chemotherapy combined with immunotherapy were in 18 cases, and surgical comprehensive therapy (surgery combined with chemotherapy and immunotherapy) were in 26 cases. Univariate analysis showed that diabetes mellitus, serum CEA and CA19-9 levels, TNM stage and treatment mode were related factors affecting the prognosis of patients with pancreatic body and tail carcinoma (all P value <0.05). Multivariate analysis indicated that TNM stage ( HR=2.536) and surgical comprehensive therapy ( HR=0.285) were the independent factors affecting the prognosis of patients with pancreatic body and tail carcinoma ( P<0.05). Median survival was 25 months (95% CI 21.416-28.584) for patients with TNM stage Ⅲ+ Ⅳ pancreatic body and tail carcinoma treated with surgical comprehensive therapy, 11 months (95% CI 7.246-14.754) for patients treated with chemotherapy combined with immunotherapy, and 6 months (95% CI 3.819-8.181) for patients treated with symptomatic treatment; the median survival time of patients with surgical comprehensive therapy was significantly longer than that of patients with chemotherapy combined with immunotherapy and symptomatic treatment, and the difference was statistically significant ( P<0.05). Conclusions:TNM stage and surgical comprehensive therapy were the prognostic factors affecting the prognosis of patients with pancreatic body and tail carcinoma, and surgical comprehensive therapy may be the best choice for long-term survival of patients.

Objective:To investigate the protective effect of ulinastatin on sepsis-acute kidney injury (SA-AKI) by NF-κB signaling pathway.Methods:Total of 60 mice were randomly(random number) divided into sham group, cecal ligation puncture group (CLP group) and ulinastatin treatment group (CLP+UTI group). Ulinastatin treatment group was intraperitoneally injected with ulinastatin 50 000 U/kg once a day. 24 hours after operation, five mice were sacrificed, the kidney tissues were collected to observe renal histopathology by HE staining. The macrophage infiltration was observed by immunohistochemistry. The remaining mice in each group were used to calculate the survival rate of 7-day after operation. HK-2 cells were stimulated by LPS to obtain the SA-AKI model, and the cells were divided into control group, LPS group and LPS + UTI group. CCK-8 assay was used to detect cell viability, EdU assay was used to detect cell proliferation, and JC-1 assay was used to detect mitochondrial damage. The phosphorylation degree of NF-κB was detected by western blot. Inflammatory factors concentrations of cellular supernatant were detected by ELISA assay.Results:Compared with the sham group, the kidney tissue of mice in CLP group showed that kidney pathological obvious changed, the infiltration of macrophages increased, and the survival rate of mice decreased. CLP+ UTI group reduced the pathological changes and the infiltration of macrophages, improved the survival rate of mice. Compared with control group, LPS group obviously inhibited the cells activity and proliferation of HK-2 cells, damaged the mitochondrial membrane potential of HK-2 cells. Compared with LPS group, LPS+ UTI group attenuated the phosphorylation of NF-κB, decreased the secretion of inflammatory factors, rescued the activity and proliferation of HK-2 cells, and reduced the damage of HK-2 mitochondrial membrane potential.Conclusions:Ulinastatin can reduce mitochondrial damage, inhibit the secretion of inflammatory factors and improve the function of renal tubular epithelial cells through regulating NF-κB signaling pathway.

Objective In this study,the diversity of the TCR β chain CDR3 in peripheral blood of patients with different typical Traditional Chinese Medicine(TCM)syndromes of liver cirrhosis was analyzed by immune repertoire sequencing,the material basis and regularity of the syndromes of liver cirrhosis was discussed.Methods 20 patients with cirrhosis,including liver and gallbladder damp heat(LGHD),liver depression and spleen deficiency(LDSD),and liver and kidney yin deficiency(LKYD)were enrolled into case group,and 10 healthy patients were used as the healthy control group.DNA was extracted from peripheral blood samples,and multiplex PCR amplification of TCR β chain CDR3 was performed,followed by high-throughput sequencing of the products to analyze the diversity of the TCR β chain CDR3.Results The nt sequence numbers unique to CDR3 and aa sequence numbers unique to CDR3 of LDSD between liver cirrhosis syndromes were less than LKYD(P<0.05).Clonality,Pielous,Shannon.Index and DE50 of LGHD and LKYD had significant differences(P<0.05)between two groups,as well as the frequency of multiple fragments in V and J regions and V-J gene recombination of LGHD vs.LDSD,LGHD vs.LKYD,and LKYD vs.LDSD,respectively(P<0.05).LGHD vs.LDSD,TRBV21-1,TRBV12-4,TRBV11-1 subtype and 7 pairs of V-J recombination have statistical differences(P<0.05).LGHD vs.LKYD,TRBV10-2,TRBV7-6,TRBV5-8 subtypes and 30 pairs of V-J recombination were statistically different(P<0.05).LDSD vs.LKYD,there were statistical differences between TRBJ1-5 subtype and 18 pairs of V-J recombination(P<0.05).Conclusions The present study was conducted to find that the diversity of TCR CDR3 in liver cirrhosis syndrome is significantly different and conforms to the regularity of syndrome from excess to deficiency by explore the immunological characteristics of different TCM syndromes of liver cirrhosis,and to provide new support for the objective basis of"combination of disease and TCM syndrome"and"diagnosis and treatment".We explored the different expression patterns and specificity of adaptive immune gene rearrangement in patients with different TCM syndromes to identify different expression patterns and specific markers of adaptive immune gene rearrangements of typical TCM syndromes in liver cirrhosis.

Hepatolithiasis is a common biliary tract disease in China and other Asian countries.It has insidious onset,recurrent disease,and may lead to biliary cirrhosis or even cancer in the late stage without inter-vention.At present,surgery is considered to be the best choice for treatment.In the 21st century of minimally inva-sive surgery,traditional procedures such as partial hepatectomy,choledochotomy,cholangioenterostomy and liver transplantation can be performed with the assistance of laparoscopy and da Vinci robots,and emerging endoscopic techniques such as endoscopic retrograde cholangiopancreatography(ERCP)and percutaneous transhepatic cho-ledochoscopic lithotripsy(PTCSL)have also been developed,bringing better treatment for patients with hepatoli-thiasis.As treatment options are becoming more varied,there are also some clinical problems that need to be addressed.In this article,we would like to briefly review the current surgical treatment modalities in order to provide a theoretical basis for optimizing the treatment modalities.

OBJECTIVETo study the severe acute respiratory syndrome (SARS)-associated coronavirus genotype and its characteristics.

METHODSA SARS-associated coronavirus isolate named ZJ01 was obtained from throat swab samples taken from a patient in Hangzhou, Zhejing province. The complete genome sequence of ZJ01 consisted of 29,715 bp (GenBank accession: AY297028, version: gi: 30910859). Seventeen SARS-associated coronavirus genome sequences in GenBank were compared to analyze the common sequence variations and the probability of co-occurrence of multiple polymorphisms or mutations. Phylogenetic analysis of those sequences was done.

RESULTSBy bioinformatics processing and analysis, the 5 loci nucleotides at ZJ01 genome were found being T, T, G, T and T, respectively. Compared with other SARS-associated coronavirus genomes in the GenBank database, an A/G mutation was detected besides the other 4 mutation loci (C:G:C:C/T:T:T:T) involved in this genetic signature. Therefore a new definition was put forward according to the 5 mutation loci. SARS-associated coronavirus strains would be grouped into two genotypes (C:G:A:C:C/T:T:G:T:T), and abbreviated as SARS coronavirus C genotype and T genotype. On the basis of this new definition, the ZJ01 isolate belongs to SARS-associated coronavirus T genotype, first discovered and reported in mainland China. Phylogenetic analysis of the spike protein gene fragments of these SARS-associated coronavirus strains showed that the GZ01 isolate was phylogenetically distinct from other isolates, and compared with groups F1 and F2 of the T genotype, the isolates of BJ01 and CUHK-W1 were more closely related to the GZ01 isolate. It was interesting to find that two (A/G and C/T) of the five mutation loci occurred in the spike protein gene, which caused changes of Asp to Gly and Thr to Ile in the protein, respectively.

CONCLUSIONAttention should be paid to whether these genotype and mutation patterns are related to the virus's biological activities,epidemic characteristics and host clinical symptoms.

Objective:To explore the efficacy and safety of high-titer plasma in the treatment of pediatric patients with severe adenovirus pneumonia.Methods:The clinical data of 92 pediatric patients with severe adenovirus pneumonia admitted to pediatric intensive care unit (PICU) in Guangzhou Women and Children′s Medical Center from January 2016 to October 2019 were retrospectively collected. According to the treatment with or without high-titer plasma, the patients were divided into plasma treatment group ( n=41) and non-plasma treatment group ( n=51). The 51 patients with chest radiograph showing more than half the lungs involved were divided into plasma treatment group ( n=29) and non-plasma treatment group ( n=22). According to fever duration before plasma treatment, patients were divided into early group (≤5 days, n=5), middle group (>5-10 days, n=14), and late group (>10 days, n=22). Baseline data, therapeutic effects, and prognosis of patients in each group were analyzed with t test, non-parametric rank sum test, one-way ANOVA and chi-square test. Results:Ninety-two patients were included. There were no significant differences in age, gender, body weight, fever duration, sequential organ failure assessment, and Murray lung injury score between plasma treatment group and non-plasma treatment group before admission (all P>0.05). The proportion of patients whose temperature drop to normal within 5 days was higher in plasma treatment group than that in non-plasma treatment group (88% (36/41) vs. 69% (35/51), χ 2=4.745, P=0.029). However, there were no significant differences between the two groups in the proportions of invasive ventilator weaning within 14 days (63% (26/41) vs. 76% (39/51), χ 2=1.868, P=0.172), transfer out from PICU within 14 days (49% (20/41) vs. 69% (35/51), χ 2=3.724, P=0.054), discharge within 28 days (51% (21/41) vs. 61%(31/51), χ 2=0.846, P=0.358) and survived patients (85% (35/41) vs. 76%(39/51), χ 2=1.143, P=0.285). Among patients with severe chest radiograph, the proportions of patients whose temperature drop to normal within 5 days and survived patients were higher in plasma treatment group than those in non-plasma treatment group (86% (25/29) vs. 59% (13/22), χ 2=4.843, P=0.028; 83% (24/29) vs. 55%(12/22), χ 2=4.796, P=0.029, respectively). However, there were no significant differences between the two groups in the proportions of invasive ventilator weaning within 14 days (52% (15/29) vs. 59% (13/22), χ 2=0.274, P=0.601), transfer out from PICU within 14 days (34% (10/29) vs. 45% (10/22), χ 2=0.632, P=0.427), and discharge within 28 days (45% (13/29) vs. 45% (10/22), χ 2=0.002, P=0.964). Among early, middle and late group, the proportions of invasive ventilator weaning within 14 days were 2/5, 13/14 and 50% (11/22), respectively, with statistically significant difference (χ 2=8.119, P=0.017). There were no significant differences in the proportions of patients whose temperature drop to normal within 5 days (4/5, 14/14, 82% (18/22), χ 2=2.965, P=0.227), transfer out from PICU within 14 days (2/5, 10/14, 36%(8/22), χ 2=4.386, P=0.112), discharge within 28 days (2/5, 8/14, 50% (11/22), χ 2=0.462, P=0.794) and survived patients (4/5, 13/14, 82% (18/22), χ 2=0.966, P=0.617) in the three groups. Only one case with high-titer plasma therapy had rash in the course of infusing plasma and no other adverse reactions were observed. Conclusions:High-titer plasma can shorten the fever time and improve the proportion of survival patients in pediatric severe adenovirus pneumonia. The clinical effect of high-titer plasma is better in 5-10 days of fever course. High-titer plasma is an effective and safe treatment.