As the core vehicle for preserving and transmitting traditional Chinese medicine（TCM） academic thought and clinical experience， the establishment of a robust quality evaluation system for TCM clinical case reports is a crucial component in the current standardization and modernization of TCM. Based on the practical experience of constructing the China Clinical Cases Library of Traditional Chinese Medicine by the China Association of Chinese Medicine， this study conducted a comprehensive analysis of critical challenges， including insufficient authenticity and unfocused evaluation criteria. It proposed a three-dimensional evaluation framework grounded in the structure-process-outcome logic， encompassing three dimensions of authenticity and standardization， characteristics and advantages， application and translational impact. This framework integrated 12 key evaluation indicators in a systematic manner. The model preserved the academic characteristics of TCM syndrome differentiation and treatment， while aligning with modern scientific research standards， achieving a balance between individualized TCM experience and standardized evaluation. Concurrently， this study provided theoretical foundations and methodological guidance for evaluating the quality of TCM clinical cases， contributing significantly to the inheritance of TCM knowledge， evidence-based practice， and the reform of talent evaluation mechanisms.

ObjectiveTo observe the clinical efficacy and safety of traditional Chinese medicine （TCM） triple therapy in the treatment of spontaneous bacterial peritonitis （SBP） with damp-heat stagnation and toxin-blood stasis syndrome in liver cirrhosis patients， and to explore its potential mechanisms of action. MethodsEighty-six patients were randomly divided into the experimental group and the control group， with 43 patients in each group. Both groups received standard western medicine treatment， while the experimental group additionally received TCM triple therapy， including oral Qingre Liangxue Jiedu Decoction （清热凉血解毒汤）， retention enema with Dachengi Decoction （大承气汤）， and abdominal application of Qingre Zhitong Lishui Fomulation （清热止痛利水方） with lotus leaf. Both groups were treated for 2 weeks. Before and after treatment， various indicators were measured， such as TCM syndrome scores， ascites volume measured by abdominal ultrasound， liver function indicators including total bilirubin （TBIL）， alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and albumin （ALB）， infection markers， including neutrophil percentage （NEUT%）， C-reactive protein （CRP）， and procalcitonin （PCT）， inflammatory factors， such as tumor necrosis factor-alpha （TNF-ɑ）， interleukin-6 （IL-6）， and interleukin-10 （IL-10）， intestinal mucosal barrier function markers， including endotoxin （ET）， diamine oxidase （DAO）， D-lactic acid （D-Lac）， occludin， and zonula occludens-1 （ZO-1）， and peritoneal polymorphonuclear （PMN） cell counts at 72 hours post-treatment. ResultsA total of 82 patients were included in the final analysis， with 41 patients in each group. The total effective rate for TCM syndrome in the experimental group was 92.68% （38/41）， which was significantly higher than the 80.49% （33/41） in the control group （P＜0.05）. Compared with pre-treatment values， both groups showed significant reductions in TCM syndrome scores， ascites volume， TBIL， ALT， AST， NEUT%， CRP， PCT， TNF-α， IL-6， ET， DAO， D-Lac， Occludin， and ZO-1， with an increase in IL-10 levels and a decrease in PMN count in ascites 72 hours post-treatment （P＜0.05）. Furthermore， the experimental group outperformed the control group in all the above indicators after treatment （P＜0.05）. The disappearance time of fever and abdominal pain was shorter in the experimental group than in the control group （P＜0.01）. There were no significant changes in routine urine and stool tests， renal function， electrolytes， or electrocardiogram in either group compared with pre-treatment values. ConclusionTCM triple therapy in addition to western medicine routine treatment could significantly improves clinical symptoms in patients with liver cirrhosis and SBP with damp-heat stagnation and toxin-blood stasis syndrome， alleviates liver inflammation， improves liver function， accelerates the resolution of ascites， and increases clinical efficacy. The potential mechanism may be related to the regulation of the inflammatory response and the promotion of intestinal mucosal barrier repair.

Objective To explore the effectiveness of an integrated laparoscopic simulation training course (best essential surgical technique training, BEST) in enhancing laparoscopic peritoneal suturing techniques in surgical residents.Methods As an integrated two-stage program, the BEST course applied basic laparoscopic training system with simple molds in phase Ⅰ training, and then adopted advanced laparoscopic training system, 3D Laparoscope and ex-vivo animal models in phase Ⅱ training. The laparoscopic suturing techniques were practiced in phase Ⅱ training. From August 2021 to July 2024, surgical residents in the second year of the national standardized training program were divided into pilot and control groups based on whether they had undergone the BEST course. Two cases of laparoscopic peritoneal suture were performed by the surgical residents under supervision in the department of gastrointestinal surgery. The operative time, quality of suture, and independent completion rate were compared between the two groups.Results A total of 33 surgical residents (19 in pilot group and 14 in control group) were included in this study, and a total of 66 cases of laparoscopic peritoneal suture were performed (38 in pilot group and 28 in control group). The operative time was significantly shorter in pilot group than that in control group (15.7 min vs. 17.5 min, P=0.025). The quality of suture was significantly better in pilot group compared to control group (P=0.023). In pilot group, all peritoneal sutures were performed by residents independently, whereas in control group, 3 cases (10.7%) were assisted by the supervisor, and the independent completion rate was different significantly (P=0.039).Conclusions The BEST course can help improve surgical residents′ laparoscopic peritoneal suturing techniques and could be promoted in the national standardized training program for surgical residents.

Objective:To investigate the difference in intracranial infection rate between long-tunneled external ventricular drainage(LTEVD)and conventional external ventricular drainage(EVD),as well as the risk factors for intracranial infection.Methods:A retro-spective analysis was performed for the clinical data of 45 patients who were admitted to Department of Neurology Center,The Third Affiliated Hospital of Chongqing Medical University,from January 2020 to December 2022 and underwent EVD,among whom 13 patients underwent LTEVD(LTEVD group)and 32 patients underwent conventional EVD(EVD group).Related data were recorded for both groups,including general information,postoperative catheter-related complications,and postoperative management,to investi-gate the effect on reducing the rate of intracranial infection.According to the presence or absence of intracranial infection after surgery,the patients were divided into the infection group with 10 patients and non-infection group with 35 patients,and related clini-cal data were analyzed to investigate the risk factors for intracranial infection.Results:The LTEVD group had a significantly lower secondary infection rate of catheterization days than the EVD group[2.40‰(1/417)vs.27.19‰(9/331),P=0.009].The duration of catheterization was 14-85 days[27.00(22.50,36.50)days]in the LTEVD group and 8-22 days[9.00(8.00,11.50)days]in the EVD group,suggesting that the LTEVD group had a significantly longer duration of catheterization than the EVD group(P=0.000).The multivariate logistic regression analysis showed that the times of cerebrospinal fluid sampling was an independent risk factor for post-operative intracranial infection in patients undergoing EVD,and the use of LTEVD was a protective factor against intracranial infection after EVD.Conclusion:Compared with conventional EVD,LTEVD can safely prolong the duration of catheterization and reduce the rate of postoperative intracranial infection in patients undergoing EVD.The use of LTEVD procedure and the reduction in the times of cerebrospinal fluid sampling can reduce the risk of postoperative in-tracranial infection.

Objective To explore the interaction between long non-coding RNA growth arrest-specific 5(GAS5)and miR-135b-5p/adenomatous polyposis coli(APC)axis,and to elucidate its biological function in the proliferation,metastasis and invasion of glioma.Methods The relationship of GAS5 expression level with survival rate of glioma patients was analyzed based on CGGA database.qRT-PCR was used to detect the expression level of GAS5 in glioma tissues and cell lines.Human glioma T98 and A172 cell lines were subjected for overexpression and knockdown of GAS5,miR-135b-5p and APC by transfection.Western blotting and qRT-PCR were applied to measure the expression of related genes in glioma tissues or cell lines.CCK-8,Transwell and wound healing assays was conducted to determine the effect of GAS5 on the viability and motility of glioma cells.Moreover,dual luciferase reporter analysis were utilized to elucidate the regulatory mechanisms of GAS5 and miR-135b-5p/APC.Results Analysis on CGGA database showed that the survival rate of glioma patients with low GAS5 expression was significantly decreased(P＜0.001).Western blotting and qRT-PCR indicated that GAS5 was down-regulated in the glioma cell lines and tissues than human normal astrocytes and para-cancer tissues(P＜0.01).The results of CCK-8,Transwell and wound healing assays revealed that overexpression of GAS5 significantly inhibited the viability,invasion and migration abilities in T98 cells when compared with the cells of the vector group(P＜0.01).Dual luciferase reporter analysis displayed that there were binding sites between GAS5 and miR-135b-5p,and the expression of the two was negatively correlated(P=0.019).Further studies suggested that the effect of GAS5 on biological function in glioma cells was through targeting miR-135b-5p,and overexpression of APC reversed the promoting effect of miR-135b-5p on glioma cells(P＜0.01).Western blotting displayed that enhancing miR-135b-5p expression inhibited the expression of APC(P＜0.001),and GAS5 upregulated the expression of APC by targeting miR-135b-5p(P＜0.01).Conclusion GAS5 inhibits the progression of glioma via miR-135b-5p/APC axis,which revealing the molecular regulatory mechanism of GAS5/miR-135b-5p/APC.

Objective To explore the effect of Sangxing Zhike Formula in rats with cough vari-ant asthma(CVA)and its possible mechanism based on the cyclic adenosine monophosphate(cAMP)/cystic fibrosis transmembrane conductance regulator(CFTR)pathway.Methods SD rats were randomly divided into control group,model group,dexamethasone group(0.5 mg/kg)and low-,medium-,high-dose Sangxing Zhike Formula groups(9.6,19.2 and 38.4 g/kg)using a ran-dom number table method,with 9 rats in each group.Except for the control group,CVA rat models were established in the other groups.Rats in each group were administered the drug by gavage once a day for 14 consecutive days.The general conditions of rats in each group were observed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β)and interleukin-18(IL-18)in rat serum.Hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining were used to observe the pathological changes in rat lung and bronchial tissues,and the acid-base balance of airway surface liquid(ASL)was measured.Western blot and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)were used to detect the expression levels of pro-tein kinase A(PKA),CFTR protein and their mRNA in lung tissues of rats.Results Compared with the control group,rats in the model group showed listlessness,dull fur,slow weight gain,a significantly expanded area of alveolar septal consolidation,and a large number of inflammatory cell infiltrations around the bronchi.Compared with the model group,rats in each intervention group had better general conditions and reduced inflammatory infiltrations in lung tissues and bronchial lu-mens.Compared with the control group,the serum levels of IL-1 β and IL-18 in the model group were increased,and the pH values of ASL in the model group,low-dose Sangxing Zhike Formula group,medium-dose Sangxing Zhike Formula group,and high-dose Sangxing Zhike Formula group were all decreased,with statistically significant differences(P＜0.05).Compared with the model group,the serum levels of IL-1β and IL-18 in each intervention group were decreased,and the pH values of ASL in the dexamethasone group,medium-dose Sangxing Zhike Formula group,and high-dose Sangxing Zhike Formula group were increased,with statistically significant differences(P＜0.05).Compared with the control group,the expressions of PKA protein and PKA mRNA in the model group,low-dose Sangxing Zhike Formula group,and medium-dose Sangxing Zhike Formula group were all decreased,and the expressions of CFTR protein and CFTR mRNA in the model group and each intervention group were all decreased,with statistically significant differences(P＜0.05).Compared with the model group,the expressions of PKA protein and PKA mRNA in the dexametha-sone group and high-dose Sangxing Zhike Formula group were increased,and the expressions of CFTR protein and CFTR mRNA in the high-dose Sangxing Zhike Formula group were increased,with statistically significant differences(P＜0.05).Conclusion Sangxing Zhike Formula can im-prove the general conditions of CVA rats,regulate the acid-base balance of ASL,reduce airway in-flammatory cell infiltration andairway remodeling,and decrease the levels of inflammatory factors IL-1β and IL-18.Its mechanism may be related to the cAMP/CFTR pathway.

Objective: To investigate the therapeutic effect of bone marrow mesenchymal stem cells ( BMSCs) mod- ified by bone morphogenetic protein ( BMP) -2 gene combined with nano-hydroxyapatite ( nHA) / polyamide 66 ( PA66) on femoral nonunion in rats． Methods : Rat BMSCs were isolated and divided into the stent + BMSCs group，stent + BMSCs + rhBMP-2 group，and stent + BMSCs + Ad-BMP-2 group; human recombinant BMP-2 ( rh- BMP-2) or adenovirus-infected BMP-2 ( Ad-BMP-2) vector was transfected into BMSCs and loaded onto nHA / PA66 stent materials．Flow cytometry was used to detect that BMSCs expressed specific surface markers．Cell pro- liferation was detected by MTT assay，related bioactive factors［platelet-derived growth factor ( PDGF) ，transfor- ming growth factor-β ( TGF-β) ，vascular endothelial growth factor ( VEGF) ，fibroblast growth factor ( FGF) ，os- teocalcin ( OCN) ，osteonectin ( ON) ］were detected by ELISA，alkaline phosphatase ( ALP) activity was detec- ted，and cell growth and adhesion were observed by scanning electron microscopy ( SEM) ．In the atrophic nonun- ion model of SD rats，the stent + BMSCs + rhBMP-2 or stent + BMSCs + Ad-BMP-2 complex was implanted into the defect area，which was divided into the rhBMP-2 group and the Ad-BMP-2 group，and the therapeutic effect was e- valuated by X-ray and MicroCT． Results: The surface of the nHA / PA66 stent was smooth and porous，and BMSCs adhered well．Flow cytometry showed high expression of CD29 and CD90 and low expression of CD45 and CD34 in BMSCs．MTT showed that the cells proliferated rapidly after 72 h．Compared with the stent + BMSCs group and the stent + BMSCs + rhBMP-2 group，the ALP activity，the expressions of PDGF，TGF-β , VEGF，FGF，OCN，and ON in the stent + BMSCs + Ad-BMP-2 group were significantly up-regulated ( P＜0. 05) ．12 weeks after surgery， the stent complex in the Ad-BMP-2 group of rats was completely wrapped by newly formed cortical bone，and the surface was smooth and intact．X-ray showed that the complex in the Ad-BMP-2 group was completely wrapped by newly formed cortical bone，and the recovery degree was better than that in the rhBMP-2 group．Micro-CT results showed that compared with the rhBMP-2 group，the bone volume fraction，trabecular thickness，trabecular num- ber，bone mineral density，and bone mineral content in the Ad-BMP-2 group all increased 12 weeks after surgery ( P＜0. 05) ，and the trabecular separation level decreased ( P＜0. 05) ． Conclusion Ad-BMP-2-transfected BM- SCs combined with nHA / PA66 stent material can express bioactivity more effectively，provide a continuous and good microenvironment for the proliferation and differentiation of BMSCs，which is beneficial to promoting local os- teogenic activity and bone defect repair．

ObjectiveTo study the effects of Huangjing Jiannao granules on learning and memory abilities and cerebral blood flow in the rat model of vascular cognitive impairment （VCI） and to explore the mechanism of Huangjing Jiannao granules in the treatment of VCI. MethodSeventy-two SPF-grade male SD rats were randomly selected， with 12 rats as the sham operation group. The remaining rats were subjected to bilateral carotid artery ligation （2-VO） for the modeling of VCI. According to the randomized block design， the successfully modeled rats were grouped as follows： model， donepezil hydrochloride （0.50 mg·kg^-1）， and low-， medium-， and high-dose （2.36， 4.72， 9.44 g·kg^-1， respectively） Huangjing Jiannao granules. After 6 weeks of treatment， Morris water maze test and new object recognition test were conducted to evaluate the learning and memory abilities of the rats. After continuous gavage for 8 weeks， the cerebral blood flow was recorded by a laser microcirculation blood flow imager， and the survival and injury of hippocampal neurons were observed by Nissl staining. The expression of neuronal nuclear antigen （NeuN） in the hippocampus was detected by immunohistochemistry （IHC）. The levels of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in the serum were determined by enzyme-linked immunosorbent assay. The protein levels of phosphatidylinositol 3-kinase （PI3K）， phosphorylated protein kinase B （p-Akt）， nuclear factor-κB p65 （NF-κB p65）， and nuclear factor-κB inhibitor α （IκBα） in the hippocampus were determined by Western blot. ResultCompared with the sham operation group， the model group showed weakened learning and memory abilities （P<0.01）， reduced blood flow in the whole brain， forebrain， and hindbrain （P<0.01）， damaged neurons and reduced survived neurons in the hippocampal CA1 region （P<0.01）， down-regulated expression of NeuN （P<0.01）， elevated levels of IL-1β and TNF-α in the serum （P<0.01）， up-regulated protein levels of PI3K， p-Akt， and NF-κB p65 in the hippocampal tissue， and down-regulated protein level of IκBα （P<0.01）. Compared with the model group， medium- and high-dose Huangjing Jiannao granules improved the learning and memory abilities （P<0.05，P<0.01）. High-dose Huangjing Jiannao granules increased the blood flow in the whole brain， forebrain， and hindbrain （P<0.05，P<0.01）， and medium-dose Huangjing Jiannao granules increased the blood flow in the whole brain （P<0.05）. All the doses of Huangjing Jiannao granules increased the number of survived neurons （P<0.05，P<0.01） and up-regulated the protein level of NeuN （P<0.05，P<0.01）. Medium and high-dose Huangjing Jiannao granules lowered the level of TNF-α （P<0.05，P<0.01）， down-regulated the protein levels of PI3K， p-Akt， and NF-κB p65 （P<0.05，P<0.01）， and up-regulated the protein level of IκBα （P<0.01）. ConclusionHuangjing Jiannao granules can improve the learning and memory abilities and promote the recovery of cerebral blood flow in the rat model of VCI induced by 2-VO by regulating the expression of proteins involved in the PI3K/Akt signaling pathway， inhibiting inflammation， and reducing hippocampal neuron injury.

[Objective] To explore the cellular composition and gene expression characteristics of adrenocortical adenoma (ACA) based on single-nuclear RNA sequencing. [Methods] The postoperative resection samples of 2 ACA cases treated at our hospital during Jul.and Aug.2022 were collected.The cellular composition was isolated and identified with single-nuclear RNA sequencing, the adrenal cortical subgroups were subdivided with specific cortical cell subgroup markers, and the characteristic gene expression profile was studied. [Results] The main components of ACA were adrenal cortical cells (78.62%), endothelial cells (1.91%), fibroblasts (3.09%), macrophages (14.34%), and T cells (2.05%). Subdivision of cortical cells revealed a group of undefined cells (1.08%), in addition to zona glomerulosa (ZG) cells (1.77%), zona fasciculata (ZF) cells (94.98%), and zona reticularis (ZR) cells (2.17%). These undefined cells were characterized by high expression of intercellular adhesion molecule 1, growth arrest and DNA damage inducing proteins β, tumor necrosis factor α inducing protein 3 and CD36 molecules.According to correlation analysis of the gene expression, these undefined cells were similar to the ZF cells.DEGs enrichment analysis indicated that the metabolic process enrichment index was the highest in biological processes.Reactome GO enrichment analysis revealed that the immune system cytokine signaling response was the most significant.KEGG signaling pathway analysis showed that Th17 cell differentiation pathway was the mostly enriched.DEGs were found to be most closely related to viral infection by DO enrichment analysis. [Conclusion] This study reveals for the first time the gene expression profile characteristics of cell subset in ACA, which can provide reference to explore the mechanism of ACA.

Bone Transplantation/adverse effects* ; Transplantation, Autologous/adverse effects*