ObjectiveTo investigate the possible mechanism of Shenfuhuang Formula （参附黄配方） in prevention and treatment of epsis-associated encephalopathy from the perspective of brain genomics. MethodsC57BL/6 mice were randomly divided into sham surgery group， sepsis group， and Shenfuhuang group， with 20 mice in each group. The sepsis group and Shenfuhuang group were induced to develop sepsis by cecal ligation and puncture （CLP） procedure. At 4 hours after modelling， Shenfuhuang group were gavaged with 2.5 g/（kg·d） of Shenfuhuang Formula， 0.5 ml each time， at 12 hours intervals， for a total of 4 times after modelling. Sepsis group and sham surgery group were given 0.5 ml of purified water orally. At 48 hours after modeling， the transcriptome sequencing was used to explore the differential gene expression in the effects of Shenfuhuang Formula on the brain regions of septic mice， and real-time PCR and ELISA were later used to further validate the differential gene and proteins expression. ResultsA total of 4605 genes were differentially expressed in Shenfuhuang group compared with sepsis group， of which 2353 genes were up-regulated and 2252 genes were down-regulated. According to the results of previous publications， six key genes were screened， including serine/threonine-protein kinase （Nek1）， myelin-associated glycoprotein （Mag）， endothelial cell-specific tyrosine kinase receptor （Tek）， a disintegrin and metalloproteinase with thrombospondin motifs 20 （Adamts20）， lymphocyte antigen 86 （Ly86）， and E3 ubiquitin-protein ligase （Traip）. Further genetic and protein validation revealed that， compared to the sham surgery group， the mRNA levels and corresponding protein levels of Nek1， Mag， Tek， Adamts20， Ly86， and Traip in the brain tissue of septic mice significantly reduced （P＜0.05）. In comparison to the sepsis group， Shenfuhuang group showed significantly increased mRNA levels and corresponding protein levels of Nek1， Mag， Tek， Adamts20， Ly86， and Traip （P＜0.05）. ConclusionThe potential therapeutic targets of Shenfuhuang Formula for treating sepsis-associated encephalopathy may be related to the Nek1， Mag， Tek， Adamts20， Ly86， and Traip genes and their encoded proteins.

ObjectiveTo explore the material basis of the "interaction of blood stasis and toxin" mechanism in cerebral ischemia-reperfusion injury， as well as the protective role of Huoxue Huayu Jiedu prescription （HXHYJDF） against ferroptosis. MethodsSixty SPF-grade male SD rats were randomly divided into six groups： sham group， model group， deferoxamine （DFO） group （100 mg·kg^-1）， low-dose HXHYJDF group （4.52 g·kg^-1）， medium-dose HXHYJDF group （9.04 g·kg^-1）， and high-dose HXHYJDF group （18.07 g·kg^-1）， with ten rats in each group. Except for the sham group， the other groups were used to replicate the model of focal cerebral ischemia-reperfusion in the middle cerebral artery of rats by the reforming Longa method. Neurological function was assessed at 1^st， 3^rd， 5^th， and 7^th days post-reperfusion using the modified neurological severity scores （m-NSS）. Brain tissue pathology and the morphology of mitochondria were observed using hematoxylin-eosin （HE） staining and transmission electron microscopy. The contents of malondialdehyde （MDA）， glutathione （GSH）， divalent iron ions （Fe²⁺）， and reactive oxygen species （ROS） in the ischemic cerebral tissue were detected using enzyme-linked immunosorbent assay （ELISA）. Immunohistochemistry and Western blot （WB） were used to detect the expression of iron death marker proteins glutathione peroxidase 4 （GPX4）， ferroportin-1 （FPN1）， transferrin receptor protein 1 （TfR1）， and ferritin mitochondrial （FtMt） in brain tissue. ResultsCompared with the sham group， the mNSS score of the model group was significantly increased （P<0.01）. HE staining showed that the number of neurons in the cortex of brain tissue was seriously reduced， and the intercellular space was widened. The nucleus was fragmented， and the cytoplasm was vacuolated. The results of transmission electron microscopy showed that the mitochondria in the cytoplasm contracted and rounded， and the mitochondrial cristae decreased. The matrix was lost and vacuolated， and the density of the mitochondrial bilayer membrane increased. The results of ELISA showed that the content of GSH decreased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS increased significantly （P<0.01）. The results of immunohistochemistry and WB showed that the expression of GPX4 and FPN1 proteins was significantly decreased （P<0.01）， and the expression of FtMt and TfR1 proteins was significantly increased （P<0.01）. Compared with those of the model group， the m-NSS scores of the high-dose and medium-dose HXHYJDF groups began to decrease on the 3^rd and 5^th days， respectively （P<0.05， P<0.01）. The results of HE and transmission electron microscopy showed that the intervention of HXHYJDF improved the pathological changes of neurons and mitochondria. The results of ELISA showed that the content of GSH in the medium-dose and high-dose HXHYJDF groups increased significantly （P<0.01）， and the contents of MDA， Fe²⁺， and ROS decreased significantly （P<0.05， P<0.01）. The content of GSH in the low-dose HXHYJDF group increased significantly （P<0.01）， and the contents of MDA and ROS decreased significantly （P<0.01）. The results of immunohistochemistry showed that the expression of GPX4 and FPN1 in the high-dose HXHYJDF group increased significantly （P<0.01）， and the expression of FtMt and TfR1 decreased significantly （P<0.01）. The expression of GPX4 and FPN1 in the medium-dose HXHYJDF group increased significantly （P<0.05）， and the expression of TfR1 decreased significantly （P<0.01）. WB results showed that the expression levels of FPN1 and GPX4 proteins in the high-dose， medium-dose， and low-dose HXHYJDF groups were significantly up-regulated （P<0.01）， and the expression levels of FtMt and TfR1 proteins were significantly down-regulated （P<0.01）. ConclusionHXHYJDF can significantly improve neurological dysfunction symptoms in rats with cerebral ischemia-reperfusion injury， improve the pathological morphology of the infarcted brain tissue， and protect the brain tissue of rats with cerebral ischemia-reperfusion injury to a certain extent. Neuronal ferroptosis is involved in cerebral ischemia-reperfusion injury， with increased levels of MDA， Fe²⁺， ROS， and TfR1 and decreased levels of FtMt， FPN1， GPX4， and GSH potentially constituting the material basis of the interaction of blood stasis and toxin mechanism in cerebral ischemia-reperfusion injury. HXHYJDF may exert brain-protective effects by regulating iron metabolism-related proteins， promoting the discharge of free iron， reducing brain iron deposition， alleviating oxidative stress， and inhibiting ferroptosis.

Objective·To compare the prognostic effects of radical resection of esophageal cancer,concurrent chemoradiotherapy and simple follow-up observation on the prognosis of patients with T1b invasion of superficial esophageal squamous cell carcinoma after endoscopic submucosal dissection(ESD).Methods·From May 2016 to May 2021,the clinical data of 67 patients with esophageal squamous cell carcinoma who were pathologically confirmed as pT1b after ESD and treated in Shanghai Chest Hospital were retrospectively analyzed.According to the additional treatment after ESD,the patients were divided into additional surgery group(S group),chemoradio-therapy group(CRT group)and observation group(O group).χ2 test was used to compare the clinical baseline data and pathological information of the three groups of patients.The Kaplan-Meier survival curve and log-rank test were used to compare the disease free survival(DFS)and recurrence free survival(RFS)of the three groups of patients,and the Cox proportional hazards regression model was used on DFS and RFS by univariate and multivariate analysis.Results·Among all 67 patients,there were 23 cases in the S group,19 cases in the CRT group,and 25 cases in the O group.There was no significant difference in age(P=0.080),gender(P=0.078),tumor length(P=0.485),tumor location(P=0.655),lesion circumferential ratio(P= 0.310),histological grading(P=0.084),depth of tumor invasion(P=0.066)and lymphovascular invasion(P=0.279)among the three groups.During(42.6±16.7)months of follow-up,tumor recurrence was observed in 10 cases(14.9%),including 6 patients(60%)with local recurrence,2 patients(20%)with regional lymph recurrence and 2 patients(20%)with distant metastasis.The median recurrence time of group S,group CRT,and group O was 40.1,36.6,and 22.1 months,and the 3-year DFSs were 100%,89.5%,and 74.5%(P-trend=0.040).Multivariate Cox analysis showed that additional esophagectomy was the key to improving independent protective factors of RFS(HR=0.097,95%CI 0.010?0.956,P=0.046).Conclusion·For patients with superficial esophageal squamous cell carcinoma confirmed as pT1b after ESD,additional surgery can significantly reduce the possibility of long-term recurrence.

Objective:To assess the impact of reducing the mesh density on the overall volume and trueness of single-crown restora-tions.Methods:15 single-crown restorations were included,and their mesh density in 3D software was simplified by 75％,50％,25％,10％and 5％respectively.The overal volume,file size and mesh edge length of the restorations with different reduction levels were compared.Deviation analysis was conducted by curving clipping the area of the shoulder,tissue surface(except for the shoulder)and occlusal surface of the restorations and compared with those of the corresponding regions in the non-simplified restorations.Trueness was assessed using the root mean square(RMS)values for the volume and trueness of different areas.Results:Reducing the mesh den-sity did not significantly affect the overal volume of the single-crown restorations(P＞0.05),but it had a significant impact on the file size(P＜0.001),mesh edge length(P＜0.001)and the trueness of the shoulder(P＜0.001),tissue surface(except for the shoulder)(P＜0.001)and occlusal surface(P＜0.001)areas.Conclusion:Reducing the mesh density had little impact on the overall volume of the restorations but significantly affected their trueness.

Objective:To improve the teaching quality of Medical Microbiology by optimizing the teaching method,adjusting the teaching content and reforming the assessment model.Methods:The students of grade 2020 and 2021 of the same major were divided into the control group and the reform group.The control group received the traditional teaching method.The reform group received the"online + offline"blended teaching method,which integrates online learning resources and ideological and political education into the theoretical content of the curriculum.And the whole process assessment system was applied to the teaching method.The teaching quality was evaluated by the whole process examination results and questionnaire survey.Results:Compared with the control group,the score in the reform group was significantly improved(P＜0.01).Results of the questionnaire survey showed that students'satisfaction with the mixed teaching method reached 97.5% .The integration of hot issues of microbiology and curriculum ideological and political education significantly improved students'learning interest,and more students wanted to engage in the work of microbiology related fields in the future.Conclusion:The practice results show that optimizing the teaching method,adjusting the teaching content and reforming the assessment mode can stimulate the students'learning interest,improve the students'independent learning ability and improve the teaching quality.

Objective:To study the correlation of uridine diphosphate glucuronosyl transferase 1A1(UGT1A1) gene polymorphism and neonatal hyperbilirubinemia among Dai ethnic group in Yunnan province.Methods:From January 2020 to December 2022, Dai neonates with unexplained neonatal hyperbilirubinemia admitted to three hospitals in Yunnan Province were prospectively enrolled into the hyperbilirubinemia group. Neonates hospitalized during the same period without hyperbilirubinemia were selected as the control group. The serum total bilirubin levels were measured and UGT1A1 gene sequencing was performed in all neonates. The differences of genotype frequency and allele frequency of UGT1A1 gene in the two groups were analyzed. Logistic regression analysis was used to analyze the effects of each mutation on hyperbilirubinemia among Dai neonates.Results:A total of 92 neonates were in the hyperbilirubinemia group and 86 in the control group. No significant differences existed between the two groups on following items: gender, age at admission, gestational age (GA), birth weight (BW), feeding pattern, white blood cell count (WBC) and hemoglobin level ( P>0.05).Three mutation loci were detected in the hyperbilirubinemia group (c.211G>A, c.1091C>T and c.1456T>G), with frequencies 45.7%, 3.3% and 2.2%, respectively. Two mutation loci were detected in the control group (c.211G>A and c.1091C>T), with frequencies 17.4% and 1.2%. Correlation analysis showed that c.211G>A frequency (45.7%) and A allele frequency (23.9%) in the hyperbilirubinemia group were significantly higher than the control group ( P<0.05). No significant differences existed in the frequencies of c.1091C>T and c.1456T>G between the two groups ( P>0.05). Logistic regression analysis showed that c.211G>A was risk factor of unexplained neonatal hyperbilirubinemia in Dai neonates ( OR=3.976, 95% CI 1.991-7.941). Conclusions:The most common mutation of UGT1A1 gene in Chinese Dai neonates with unexplained neonatal hyperbilirubinemia in Yunnan Province is c.211G>A, which increases the risk of hyperbilirubinemia.

Objective Explore changes in polysaccharides in Jiangxiangru before and after ginger juice preparation,and evaluate polysaccharide anti-inflammatory and antioxidant activities before and after processing.Methods The contents of Jiangxiangru polysaccharide(JXRPs)and Ginger juice processed of Jiangxiangru polysaccharide(JZJXRPs)before and after processing were determined by phenol-sulfuric acid method.We used the swelling model in rats and endotoxin(LPS)to establish the RAW264.7 mouse macrophage inflammation model.The optimal administration concentration was determined using a cell proliferation(MTT)assay.Enzyme-linked immunosorbent assay(ELISA)were used to measure Interleukin-6(IL-6),Interleukin-12(IL-12),Nitric oxide(NO),Interleukin-4(IL-4),and Interleukin-10(IL-10).Bleeding time of mice by tail cutting was observed to evaluate the hemostatic effect.The ability to remove 1,1-diphenyl-2-picryl-hydrazyl radical(DPPH)and 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)(ABTS)was used to evaluate in vitro antioxidant activity.Results The contents of JXRPs and JZJXRPs were 13％and 22％,respectively.In swollen rats at 4 h after injection,compared with the model group,200 mg/kg JXRPs and 100 mg/kg JZJXRPs significantly reduced rat swelling(P＜0.05).In vitro anti-inflammation assessment showed that the polysaccharides before and after processing significantly inhibited secretion of IL-6,IL-12,and NO(P＜0.01)and promoted secretion of IL-4 and IL-10(P＜0.01),and also that processing post-effects were stronger.The hemostatic experiment show that,compared with the control group,JXRPs increased hemostasis,but without a significant difference,and no significant difference was found using JZJXRPs,although high doses showed a trend to increase hemostasis.In vitro antioxidant activity showed that JXRPs and JZJXRPs had different scavenging abilities for DPPH and ABTS with IC50 values of JXRPs of 0.2215 and 0.2110 mg/ml,respectively,and IC50 values of JZJXRPs of 0.1651 and 0.1884 mg/mL,respectively.Conclusions After Jiangxiangru is produced in ginger juice,it promotes dissolution of polysaccharides and increases polysaccharide content.Anti-inflammatory,hemostasis,and antioxidant capacities are stronger in JZJXRPS than JXRPS,which lays the foundation for follow-up research and clinical applications of JXRPS.

Objective:To systematically evaluate the efficacy and safety of ruxolitinib in the treatment of Chinese patients with refractory graft-versus-host disease(SR-GVHD) by using meta-analysis.Methods:China National Knowledge Infrastructure(CNKI), WanFang database, VIP database, China Biology Medicine disc, PubMed, Ebscore Medline, EMBASE, Web of Science Core Collection and Cochrane Library were searched by using "ruxolitinib" "Graft vs Host Disease" "graft versus host disease" "Graft-Versus-Host Disease" as key words. The retrieve time was from the establishment of the database to December 7th, 2021. The related literatures of ruxolitinib for Chinese patients with SR-GVHD were screened according to inclusion and exclusion criteria, and the characteristics of the literatures were extracted. Outcomes indexes included overall objective response rate (ORR), proportion of hormone reduction and discontinuation, survival indexes and incidence of adverse events. Meta-analysis of ORR, remission rate of affected organs, 12-month overall survival rate, overall mortality and mortality due to graft-versus-host disease (GVHD) were performed by using Stata 16.0 software or R3.6.3 software. The statistics analysis was performed on the hormone usage and adverse events.Results:A total of 19 literatures involving 775 Chinese patients with SR-GVHD treated by ruxolitinib were included. Meta analysis showed that the ORR of ruxolitinib for treatment of acute GVHD (aGVHD) was 84% (285/339), with moderate heterogeneity among studies ( I2 = 62.04%, P<0.01), and the complete remission (CR) rate, partial remission (PR) rate and non-remission (NR) rate was 56% (190/339), 28% (95/339), and 14%(47/339), respectively. The ORR of ruxolitinib for treatment of chronic GVHD (cGVHD) was 77% (332/431), with moderate heterogeneity among studies ( I2 = 50.17%, P = 0.02), and the CR rate, PR rate and NR rate was 36% (155/431), 41% (177/431) and 21% (91/431), respectively. As for hormone usage, 46.51% of aGVHD patients had steroid reduction and 34.88% patients had steroid discontinuation. Similarly, 28.2% of patients with cGVHD had steroid reduction and 36.9% had steroid discontinuation. The overall 12-month survival rate of patients with SR-GVHD after the treatment of ruxolitinib was 71% (95% CI: 63%-79%, I2 = 72.70%, P<0.01). Adverse events occurred for 760 times in total, among which 287 times (36.0%) of viral infection and 269 times (34.7%) of hemocytopenia were the most common adverse events. Conclusions:Ruxolitinib is effective in the treatment of Chinese patients with SR-GVHD, and it helps reduce hormone-dependency and prolong the survival time.

Objective To compare paired boxed gene 1(PAX1)methylation and p16/Ki-67 double staining alone and in combination in thinprep cytologic test(TCT)for distinguishing atypical squamous cells of undetermined significance(ASC-US)and low-grade squamous intraepithelial lesion(LSIL)population.Methods A total of 247 patients with TCT results of ASC-US and LSIL admitted in our hospital from January 2021 to December 2022 were enrolled in this study.Detection efficacy of PAX1 methylation and p16/Ki-67 double staining alone and in combination was evaluated with colposcopic pathologic results as the gold standard and the sensitivity,specificity,accuracy and area under the curve(AUC)as evaluation indexes.Results The positive rates of PAX1 methylation and p16/Ki-67 double staining were increased with the severity of pathological findings.Combined detection of PAX1 methylation and p16/Ki-67 assay had a sensitivity of 91.25％,specificity of 97.72％and accuracy of 95.51％in the women with TCT of ASC-US,and these values were statistically better than those of PAX1 methylation and p16/Ki-67 double staining alone(P＜0.01).Conclusion The combination of PAX1 methylation and p16/Ki-67 double-staining assay can further improve the diagnostic efficacy in patients with ASC-US on TCT results.

Objective To investigate the feasibility and clinical experience of kidney transplantation from donors with Marfan syndrome (MFS). Methods Clinical data of 2 recipients undergoing kidney transplantation from the same MFS patient were retrospectively analyzed and literature review of 2 cases was conducted. Characteristics and clinical diagnosis and treatment of kidney transplantation from MFS patients were summarized. Results The Remuzzi scores of the left and right donor kidneys of the MFS patient during time-zero biopsy were 1 and 2. No significant difference was observed in the renal arteriole wall compared with other donors of brain death and cardiac death. Two recipients who received kidney transplantation from the MFS patient suffered from postoperative delayed graft function. After short-term hemodialysis, the graft function of the recipients received the left and right kidney began to gradually recover at postoperative 10 d and 20 d. After discharge, serum creatinine level of the recipient received the left kidney was ranged from 80 to 90 μmol/L, whereas that of the recipient received the right kidney kept declining, and the lowest serum creatinine level was 232 μmol/L before the submission date (at postoperative 43 d). Through literature review, two cases successfully undergoing kidney transplantation from the same MFS donor were reported. Both two recipients experienced delayed graft function, and then renal function was restored to normal. Until the publication date, 1 recipient has survived for 6 years, and the other recipient died of de novo cerebrovascular disease at postoperative 2 years. Conclusions MFS patients may serve as an acceptable source of kidney donors. However, the willingness and general conditions of the recipients should be carefully evaluated before kidney transplantation. Intraoperatively, potential risk of tear of renal arterial media should be properly treated. Extensive attention should be paid to the incidence of postoperative complications.