Daochisan is a pediatric famous classical formula in traditional Chinese medicine（TCM） specializing in clearing the heart， disinhibiting water and enriching Yin， which has been inherited and developed by medical experts of successive generations. The formula can be traced back to Xiaoer Yaozheng Zhijue written by QIAN Yi from Song dynasty， and most of the later-developed Daochisan（Daochitang） is based on this formula with additions and subtractions. So as to promote the literature excavation of this formula， this paper conducts a thorough textual research and analysis of the composition， processing， preparation methods， clinical applications of Daochisan based on a systematic collation of relevant ancient and modern literature in the method of bibliometrics. A total of 127 pieces of data involving 86 antient TCM books were collected. Statistical analysis showed that the drug origin of Daochisan is basically clear， we suggest that Rehmanniae Radix is the root tuber of Rehmannia glutinosa of Scrophulariaceae， Akebiae Caulis is the dry stem of Akebia quinata of Lardizabalaceae， Glycyrrhizae Radix et Rhizoma is the dry root and stem of Glycyrrhiza uralensis of legume， bamboo leaf is the dry leaves of Phyllostachys nigra var. henonis of gramineous plants. Four herbs are recommended to use its raw products. The key information of the dosage and decocting method is supposed to be "4.13 g each of raw Rehmanniae Radix， raw Glycyrrhizae Radix et Rhizoma， Akebiae Caulis， adding 300 mL of water， and then adding 4.13 g of bamboo leaf， decocting together to 150 mL， taking warmly after meal". Daochisan has the effect of clearing the heart and enriching Yin， disinhibiting water and relieving stranguria， and is widely used in treating symptoms such as heart vexed hotness， hydrodipsia and reddish complexion. Modern studies have showed that Daochisan are frequently used in treating oral ulcer， herpetic stomatitis， urinary tract infection， herpes zoster and other diseases. The above research results can provide scientific reference for the future development and research of Daochisan.

BACKGROUND:Autoimmune regulator gene(Aire)and Wnt signaling pathway play an important role in the maintenance and differentiation of mouse embryonic stem cell pluripotency.However,whether the Wnt signal and Aire are involved in the differentiation of embryonic stem cells to thymic epithelial progenitor cells remains poorly understood. OBJECTIVE:To investigate the relationship of the Wnt signaling pathway and Aire with the differentiation of embryonic stem cells. METHODS:A two-step differentiation method was used to induce mouse embryonic stem cells to differentiate into endoderm and then into thymic epithelial progenitor cells.Mouse embryonic stem cells were infected with Aire shRNA lentivirus,and monoclonal stable strains were screened by puromycin.Mouse embryonic stem cells were collected on days 0,3 and 10 of the directed induction of differentiation after the induced differentiation by the two-step differentiation method.Cellular immunofluorescence,flow cytometry,western blot assay,and real-time qPCR were used to detect the expression changes of related genes and proteins. RESULTS AND CONCLUSION:(1)Immunofluorescence staining showed positive expression of SSEA1 and OCT4 on day 0 of targeted induction of differentiation.(2)Immunofluorescence staining showed double-positive expression of SOX17 and FOXA2 on day 3 of targeted induction of differentiation.(3)Flow cytometry results showed positive expression of EPCAM1,K5 and K8 on day 10 of targeted induction of differentiation.(4)Compared with undifferentiated mouse embryonic stem cells,the expressions of Wnt7a,β-catenin,and Gsk-3β proteins were elevated,and the expression level of Aire protein was decreased in induced differentiated thymic epithelial progenitor cells.(5)Compared with undifferentiated mouse embryonic stem cells,the expressions of Wnt7a,β-catenin,Gsk-3β and Aire mRNA were elevated in thymic epithelial progenitor cells.(6)Compared with normal cultured mouse embryonic stem cells and their ultimately differentiated thymic epithelial progenitor cells,the expression levels of Wnt7a,β-catenin and Gsk-3β proteins were reduced in mouse embryonic stem cells with knockdown of Aire genes and their final differentiated thymic epithelial progenitor cells.In conclusion,the Wnt signaling pathway and Aire are jointly involved in the process of targeted induction of differentiation of mouse embryonic stem cells into mouse thymic epithelial progenitor cells.

ObjectiveBased on non-targeted metabolomics， to analyze the regulation of endogenous differential metabolites in serum of type 2 diabetes mellitus（T2DM） rats by Fuling Yunhua granules， and to clarify the metabolic pathways through which this granules exerted its effect on improving T2DM. MethodSeventy SD rats， half male and half female， were randomly divided into the control group， model group， and high， medium， low dose groups of Fuling Yunhua granules（20.70， 10.35， 5.18 g·kg^-1 in raw drug amount） and the positive drug group（pioglitazone hydrochloride tablets， 8.1 mg·kg^-1）. Except for the control group， other groups were fed with high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin（STZ） to establish a T2DM rat model. After successful modeling， the treatment groups were administered the corresponding drugs by gavage， and the control group and model group were treated with an equal volume of saline by gavage， once/d， for 28 d. Fasting blood glucose（FBG） and glycosylated hemoglobin A1c（GHbA1c） levels were measured in all groups of rats during the administration period， and hematoxylin-eosin（HE） staining was used to observe the pathomorphological changes in the pancreatic tissues of rats at the end of the administration period. The endogenous metabolite levels in rat serum were detected by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-LTQ-Orbitrap MS）， and the data were processed using principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Differential metabolites were identified by the Human Metabolome Database（HMDB） and the Kyoto Encyclopedia of Genes and Genomes（KEGG）， and screened for differential metabolites with variable importance in the projection（VIP） value>1， P<0.05， and fold change（FC）<0.6 or FC>1. And the metabolic pathway enrichment analysis of the screened differential metabolites was performed by MetaboAnalyst 5.0， then the screened differential metabolites were diagnosed and evaluated by the receiver operating characteristic（ROC） curves. ResultCompared with the control group， the FBG level of rats in the model group increased significantly（P<0.01）， the GHbA1c content tended to increase， but the difference was not statistically significant， and the pancreatic tissue of rats was obviously damaged， the number of pancreatic islets decreased， and the pancreatic β-cells were obviously reduced， atrophied and enlarged. Compared with the model group， the FBG levels of rats in the high dose group of Fuling Yunhua granules and the positive drug group were significantly reduced after 2 weeks of administration（P<0.05， P<0.01）， the GHbA1c content of rats in the high dose group of Fuling Yunhua granules was significantly reduced（P<0.05）， and the pancreatic tissue lesions of rats in the different dose groups of Fuling Yunhua granules were reduced. The results of non-targeted metabolomics showed that 46 differential metabolites were significantly changed in the model group compared with the blank group. Pathway enrichment analysis found that T2DM mainly affected biological processes including biosynthesis of primary bile acid， D-amino acid metabolism， steroid hormone biosynthesis， and glycerophospholipid metabolism in rats. Compared with the model group， the levels of 8 differential metabolites in the high dose group of Fuling Yunhua granules were significantly adjusted， and the pathway enrichment analysis found that D-amino acid metabolism， retinol metabolism， glycine， serine and threonine metabolism， tryptophan metabolism and other metabolic pathways were mainly involved. ROC curves further analysis revealed that the four characteristic differential markers of 11-cis-retinol， D-piperidinic acid， D-serine， and p-cresol sulfate had high diagnostic value for the treatment of T2DM with Fuling Yunhua granules. ConclusionFuling Yunhua granules can improve the symptoms of T2DM rats by regulating the amino acid metabolic and retinol metabolic pathways through the modulation of endogenous differential metabolites.

Objective:To explore the advantages of minimally invasive internal fixation assisted by an orthopedic robot in the treatment of tibial plateau fractures of Schatzker types Ⅱ and Ⅲ that can be indirectly reduced.Methods:A retrospective study was conducted of the 18 patients who had been treated for tibial plateau fractures of Schatzker types Ⅱ and Ⅲ at Department of Orthopaedics, The Fourth Hospital of Wuhan from December 2019 to December 2021. They were 12 males and 6 females with an age of (45.2±9.6) years. All fractures were closed. Of them, 6 were complicated with an avulsion fracture at the insertion point of the anterior cruciate ligament, 1 with tear of the medial collateral ligament, and 8 with tear of the lateral meniscus. All patients were treated with minimally invasive internal fixation using the "fence" screw technique after indirect reduction assisted by an orthopedic surgical robot. Those combined with avulsion fracture of the anterior cruciate ligament and meniscus tear underwent one-stage arthroscopic surgery, while those combined with tear of the medial collateral ligament underwent one-stage open repair. The fracture reduction was evaluated according to the Rasmussen radiological scoring system, and the knee joint function evaluated using the American Hospital for Special Surgery (HSS) scoring system.Results:All the 18 patients were fully followed up for (10.6±1.9) months. The X-ray films immediately after surgery showed good fracture reduction. The fractures healed after (11.3±1.2) weeks. At 6 months after surgery, the Rasmussen knee score was (16.8±1.0) points, giving 5 excellent and 13 good cases; the HSS score was (93.2±3.0) points, giving 17 excellent and 1 good cases. By the last follow-up, no serious complications occurred, such as common peroneal nerve injury, popliteal vascular injury, postoperative infection, or internal fixation failure.Conclusion:Since minimally invasive internal fixation assisted by an orthopedic robot can lead to fine clinical efficacy for tibial plateau fractures of Schatzker types Ⅱ and Ⅲ, this technique can be widely applied in clinical practice.

AIM:To explore the expression and distribution of synaptic and extrasynaptic glutamate receptors under the action of amyloid β-protein 42 oligomers(Aβ42Os)in Alzheimer disease.METHODS:In vitro,primary neu-rons from neonatal mice were treated with different concentrations of Aβ42Os.In vivo,Aβ42Os were stereotaxically injected into the lateral ventricle of C57BL/6 mice.Western blot was used to detect the protein levels of metabotropic gultamate re-ceptor 5(mGluR5)and N-methyl-D-aspartate receptor(NMDAR)subunits(NR2A,NR2B and NR1)in mouse primary neurons and mice.Immunofluorescence staining was performed to observe the distribution of mGluR5 and NMDAR.Addi-tionally,Western blot was employed to examine the expression of mGluR5 downstream phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/AKT)signaling pathway-related proteins,calcium signaling pathway-related proteins and synapse-related proteins in mouse hippocampal tissues.RESULTS:(1)High concentrations of Aβ42Os increased mGluR5 expression in mouse primary neurons,but reduced the expression of NR2A,NR2B and NR1(P<0.01).Treatment of pri-mary neurons with high concentration of Aβ42Os resulted in aggregation of mGluR5 on the membrane surface,and re-duced the expression of NR2A,NR2B and NR1 on the membrane surface.(2)High concentration of Aβ42Os increased the expression of synaptic mGluR5 in mouse hippocampal tissues(P<0.01),but reduced the expression of synaptic NR2A(P<0.05)and NR2B(P<0.01)and increased the expression of extrasynaptic NR2B(P<0.01).High concentration of Aβ42Os inhibited the expression of PI3K and the phosphorylation of AKT and extracellular signal-regulated kinase(ERK),and overactivated calcium/calmodulin-dependent protein kinase IIα(CaMKIIα).High concentration of Aβ42 decreased the expression of postsynaptic density protein 95,spinophilin,synaptophysin and microtubule-associated protein 2(P<0.01).CONCLUSION:(1)High concentrations of Aβ42Os increase mGluR5 expression and decrease NR2A,NR2B and NR1 expression in synapse,along with an increase in extrasynaptic NR2B.(2)High concentrations of Aβ42Os inhibit the PI3K/AKT and ERK signaling pathways,overactivated the CaMKIIα pathway,and destroyed the synaptic structures.

Purpose To explore the differential expression of GPR110,an adherent G protein-coupled receptor,and its role in the differential diagnosis of acinar and solid adenocarcinoma of the lung.Methods The expression level of GPR110 was de-termined by immunohistochemistry(IHC),qRT-PCR and ELISA,and ROC and area under the curve(AUC)were ana-lyzed to distinguish the acinar predominant and solid predomi-nant of lung adenocarcinoma,so as to evaluate the role of differ-ential GPR110 expression in the differential diagnosis of these two histopathological subtypes with different prognosis.Results The expression of GPR110 in lung adenocarcinoma tumor tis-sue was significantly higher than that in adjacent tissue,and its expression in solid predominant lung adenocarcinoma was signifi-cantly higher than that in acinar predominant.The average con-centrations of GPR110 protein in 100 pairs of acinar predominant lung adenocarcinoma tumor tissues and its adjacent tissues were 430.53 and 313.26 ng/L by ELISA.The average concentrations of GPR110 protein in 53 pairs of solid predominant lung adeno-carcinoma tumor tissues and its adjacent tissues were 716.56 and 368.46 ng/L,and the differences were statistically signifi-cant(P＜0.001).At the same time,the ROC curve showed that the GPR110 protein had a sensitivity of 77.36％,a speci-ficity of 83.00％,an optimal Cut-off value of 582.27 ng/L,and an AUC of 0.865(0.802-0.927).Conclusion GPR110 has potential application value in the differential diagnosis of acinar type and solid type of adenocarcinoma of the lung,and it is ex-pected to become a new biomarker for differential diagnosis

Objective:To explore the economic applicability and safetyof the novel uterovaginal pubic comb suspension(UPCS)surgery with Mersilene tape in the treatment of pelvic organ prolapse(POP).Methods:A ret-rospective analysis was conducted on the clinical data of patients who underwent UPCS surgery due to POP from January 1st,2021 to February 28,2023.They were divided to the UPCS surgery with Mersilene tape group(group A)and suspension surgery with Y-shaped mesh group(group B)respectively.The POP-Q indication points,sus-pension surgery duration,intraoperative bleeding volume,material expense,postoperative catheter retention time,anal exhaust time and hospitalization duration were recorded for both groupbefore and after surgery.Evaluate the severity of POP related symptoms in patients before and after surgery using the pelvic Floor Distress Invento-ry-short Form 20(PFDI-20)and Pelvic Organ Prolapsed/Urinary Incontinence Sexual Questionnaire-12(PISQ-12),and follow up and observe the patients and analyze the complications.Results:A total of 17 POP patients were included in the study.There were 12 patients in group A while 5 patients in group B.The suspension material expense of group A was considerably lower than that of group B(P＜0.05).There was no significant difference between the two groups in preoperative PFDI-20 score,preoperative PISQ-12 score,UPCS surgery duration,intr-aoperative bleeding volume,postoperative urinary catheter retention time,postoperative anal exhaust time and hospitalization duration.All patients showed stable vital signs during the surgery and no severe complications were reported.Compared with the preoperative status,the positions of the Aa,Ba,and C indicatorpoint in group A and group B were all increased significantly(P＜0.05).The PFDl-20 and PISQ-12 scores of the two groups at the last follow-up after surgery showed significant improvement compared to those before surgery(P＜0.05).No signifi-cant difference was found in the PFDI-20 and PISQ-12 scores between the two groups after surgery(P＞0.05).There were no significant differences in the postoperative complications between the two groups(P＞0.05).Con-clusions:Compared with suspension surgery with Y-shaped mesh,UPCS surgery with Mersilene tape is safe and effective in the treatment of POP.The UPCS surgery with Mersilene tape showed better cost-effectiveness in the treatment of POP,and the surgical steps are relatively simple.Therefore,UPCS surgery with Mersilene tape was worthy of promotion in clinical practice.

ObjectiveTo monitor the microbes in urban drinking water in Jiangxi from 2016 to 2020， to analyze the change in microbial qualification rate， and to provide a scientific basis for government decision-making. MethodsAccording to the Standard Examination Method for Drinking Water （GB/T 5750‒2006） and the Standards for Drinking Water Quality （GB 5749‒2006）， the water samples were collected， tested and evaluated for hygienic safety. The chi-square test was used to compare the qualification rates among different water periods， water source types， water supply modes， water samples， treatment processes， and disinfection methods. ResultsA total of 10 584 water samples were collected and examined from 2016 to 2020，with a qualification rate of 97.72%. The qualified rate of the microbiological index increased gradually over the years. There was no statistically significant difference in the microbiological qualification rate of water samples monitored in different water periods （χ²=0.718，P=0.398）， and the qualification rates were 97.85% and 97.60% in dry and abundant water periods respectively. There was a statistically significant difference in the qualification rates of water samples monitored in different water source types （χ²=79.560，P=0.398）， with groundwater having a higher qualification rate of 98.83% than surface water （97.70%）. The microbiological pass rate of water samples differed among different water supply methods （χ²=201.836，P<0.001）. The microbiological pass rates of municipal centralized water supply， self-built centralized water supply and secondary water supply were 97.51%， 88.94% and 96.56% respectively. The microbiological pass rates of water samples differed among different treatment processes. There were differences in the qualification rates of the water samples between different treatment processes（χ²=855.034，P<0.001）， with 98.22%， 67.19%， 95.60% and 53.13% of the conventionally treated， precipitated filtered， disinfected only and untreated water samples， respectively. The differences in the microbiological qualification rates of the monitored water samples between different disinfection methods were statistically significant （χ²=904.993，P<0.001）. Chlorination disinfection， chlorine dioxide disinfection， other disinfection methods and non-disinfected water samples had the pass rates of 98.35%， 98.19%， 84.38% and 60.16%， respectively. ConclusionThe qualified rate of the microbiological index of drinking water in Jiangxi increases year by year， but there are still some drinking water without appropriate disinfectants and water quality treatment processes. There is still a need to strengthen the purification， disinfection and monitoring of drinking water.

Objective:To evaluate the clinical efficacy of multi-disciplinary single center's CCCG-HB-2016 regimen in the treatment of hepatoblastoma (HB) in children.Methods:Clinical data of 36 HB patients treated with CCCG-HB-2016 program from Aug 2016 to March 2020 were analyzed.Results:These 36 patients included 20 boys and 16 girls. The serum AFP was all higher than 2 792 ng/ml,there was a correlation between AFP and tumor risk stratification ( H=14.973, P<0.05). Twenty eight cases (77.78%) were epithelial type and 8 cases (22.22%) were mixed epithelial mesenchymal type.All children were treated by tumor resection combined with chemotherapy, and there was a correlation between tumor risk stratification and surgical resection of liver lobe ( H=8.847, P<0.05). The probability of bone marrow suppression in the low-risk group was 58.33% (35/60),that in the intermediate-risk group was 73.49% (61/83) and in the high-risk group was 80.23% (69/86).All 36 cases were followed up to March 31, 2020,with an average follow-up of 21.9 months and the median survival was 22.5 months.The overall survival rate (OS) and event-free survival rate (EFS) were 97.2% and 83.3% respectively. Conclusions:The multidisciplinary CCCG-HB-2016 regimen was with a high success rate and along with a high incidence of bone marrow suppression.

Objective: To explore the new gene mutation and clinical manifestations of Caroli syndrome in 2 blood brother cases. Methods: The clinical data of the 2 brothers with clinical diagnosis of Caroli syndrome were analyzed, and the related genes were detected by second-generation sequencing technology. Results: The PKHD1 gene of both brothers had the mutation of c. 5059A>T(exon32). c. 4358(exon 32)delC shift mutation, neither of which has been reported in OMIM database. Conclusion After analysis, c. 4358(exon32) delC and c. 5059A>T(exon32) may be the new pathogenic mutation of this disease, with the same mutant genotype but completely different phenotype.