To thoroughly implement the strategic deployment outlined in the Opinions of the Central Committee of the Communist Party of China and the State Council on Promoting the Inheritance and Innovative Development of Traditional Chinese Medicine regarding research on dominant diseases of traditional Chinese medicine and to uphold the development philosophy of equal emphasis on traditional Chinese medicine and western medicine，the China Association of Chinese Medicine has fully played a leading academic role by systematically organizing and conducting a series of academic youth salons on clinical dominant diseases of traditional Chinese medicine. On September 13，2024，the 36^th Youth Salon on Clinical Dominant Diseases was successfully held in Nanjing，focusing on the advantages of traditional Chinese medicine and the integrative traditional Chinese medicine and western medicine in the diagnosis and treatment of erectile dysfunction （ED）. The conference brought together leading experts from traditional Chinese medicine，western medicine，and interdisciplinary fields，facilitating in-depth multidisciplinary discussions that led to key consensus on optimizing traditional Chinese medicine treatment protocols for ED，researching and developing new drugs of traditional Chinese medicine，and advancing interdisciplinary development in traditional Chinese medicine. This salon systematically sorted out the clinical strengths and distinctive features of traditional Chinese medicine in the diagnosis and treatment of ED. Based on current research foundations and clinical needs，it identified key directions for future scientific layout and scientific research tackling： （1） Standardization of syndrome differentiation system of traditional Chinese medicine for ED. （2） Optimization and standardization of intervention methods of integrated traditional Chinese medicine and western medicine. （3） High-quality clinical research guided by evidence-based medicine. （4） In-depth analysis of the pharmacological mechanisms of traditional Chinese medicine in the treatment of ED. （5） Clinical translation and application promotion of new drugs of traditional Chinese medicine. （6） Interdisciplinary integration and innovation in traditional Chinese medicine. For each research direction，key focus areas，expected objectives，and clinical value were further refined，along with the establishment of a scientifically sound priority funding level evaluation system. Therefore，building on the series of salons on the ED-focused dominant diseases of traditional Chinese medicine，this paper provides standardized guidance for clinical practice of traditional Chinese medicine in ED management，effectively contributing to the high-quality development of traditional Chinese medicine. It serves as a valuable reference for national scientific and technological strategic layout， research and development decision-making in new drugs of traditional Chinese medicine，research topic planning，and clinical guideline formulation.

Hydroxyectoine, a vital compatible solute, is widely utilized in cosmetics, food, pharmaceutical industries, and biologics. However, the current microbial fermentation methods for hydroxyectoine production face challenges including insufficient precursor supply and low yields. Therefore, developing engineering microbial strains capable of efficiently synthesizing hydroxyectoine is of great significance. In this study, we first constructed a high-yield ectoine-producing strain ECT04 by multi-copy integration of the ectoine synthesis genes ectABC into the pseudogene loci of Escherichia coli MG1655(DE3), achieving an ectoine titer of 6.03 g/L. Subsequently, we employed plasmids with varying copy numbers to express ectD from Chromohalobacter salexigens to enable the conversion for hydroxyectoine production. We further investigated the effects of promoter, co-substrate ɑ-ketoglutarate, Fe²⁺ concentration, and dissolved oxygen on hydroxyectoine synthesis. Through fed-batch fermentation in a 7-L bioreactor, we significantly enhanced the hydroxyectoine production efficiency, attaining a final titer of 8.58 g/L and a productivity of 0.24 g/(L·h). This work successfully achieved the de novo synthesis of hydroxyectoine in E. coli, laying a foundation for the efficient bioproduction of this compound.
Escherichia coli/genetics* ; Fermentation ; Amino Acids, Diamino/biosynthesis* ; Bioreactors/microbiology* ; Metabolic Engineering/methods* ; Chromohalobacter/genetics* ; Plasmids/genetics*

Objective To explore the efficiency improvement in segmenting neural network with the application of Transformer + U-Net artificial intelligence (AI) and modeling with the application of Python scripts in three-dimensional (3D) printing brachytherapy. Methods A Transformer + U-Net AI neural network model was constructed, and Adam optimizer was used to ensure rapid gradient descent. Computed tomography or magnetic resonance imaging data of patients were standardized and processed as self-made data sets. The training set was used to train AI and the optimal result weight parameters were saved. The test set was used to evaluate the AI ability. Python programming language was used to write an automated script to obtain the output segmentation image and convert it to the STL file for import. The source applicator and needle could be automatically modeled. The time of automatic segmentation and modeling and the time of manual segmentation and modeling were entered by two people, and the difference was verified by paired t-test. Results Dice similarity coefficient (DSC), mean intersection over union (MIOU), and Hausdorff distance (HD95) were used for evaluation. DSC was 0.9341, MIOU was 0.8762, and mean HD95 was 2.516. The mean time of manual segmentation was 1187 s, and the mean time of AI segmentation was 145 s (P < 0.01). The mean time of manual modeling was 321 s, and the mean time of modeling with Python automated scripts was 18 s (P < 0.01). Conclusion The application of Transformer + U-Net automatic segmentation and Python automated scripts can effectively improve work efficiency in the 3D printing brachytherapy for cervical cancer.

Objective　To investigate the role of granulocyte-macrophage colony-stimulating factor (GM-CSF) deficiency in the pathogenesis of Exophiala oligosperma (E. oligosperma) infection, a dematiaceous fungus, aiming to provide new insights and evidence for the treatment of dematiaceous fungal infections. Methods　C57BL/6 wild-type (WT) mice and Csf2 gene knockout (KO) mice (C57BL/6 background) were selected. Using E. oligosperma isolated from patients with caspase recruitment domain-containing protein 9 (CARD9) gene deficiency, a murine subcutaneous infection model was established to simulate the human infection route. The natural progression of infection in the mice was observed for six weeks, with skin lesion tissues collected at appropriate time points for pathological analysis and monitoring immune responses. Results　Both WT and Csf2 KO mice exhibited spontaneous pathogen clearance and gradual recovery of foot tissue appearance during the progression of infection, with a 100% survival rate at the end of observation. Compared to WT mice, Csf2 KO mice showed reduced footpad swelling at 1 and 2 weeks post-infection (t=4.674, t=5.961, P<0.01). Fungal clearance in Csf2 KO mice was delayed, with fungal colonies still detectable in lesion tissues at week 4, and Periodic Acid-Schiff (PAS)-positive fungal spores observed in histopathological sections. There was no significant difference found in macrophage infiltration between WT and Csf2 KO mice during the early stages of infection (1-2 weeks) (P>0.05), while neutrophil infiltration was significantly reduced in Csf2 KO mice at week 2 (t=3.287, P<0.01). In addition, Csf2 KO mice exhibited lower levels of IL-6 and IL-1β in foot lesion homogenates at week 1 (t=4.686, t=4.102, P<0.05). Conclusions　This study demonstrated that GM-CSF deficiency delays pathogenic fungi clearance, prolongs the disease course and affects early inflammatory cytokine production as well as neutrophil infiltration during the early stages of fungal infections.

Objective To investigate the effect of human umbilical vein endothelial cells(HUVECs)on the proliferation and stem-ness of human dental pulp stem cells(hDPSCs)silencing with integrin α6(ITGA6).Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs,and its silencing efficiency was detected.Then HUVECs were added to the chambers to co-culture,and the experiments were divided into four groups(sh-NC,sh-ITGA6,sh-NC+HUVECs and sh-ITGA6+HUVECs).hDP-SCs in the sh-NC and sh-ITGA6 groups were transfected with sh-NC and sh-ITGA6 respectively.hDPSCs transfected with sh-NC and sh-ITGA6 were co-cultured with HUVECs in the sh-NC+HUVECs group and sh-ITGA6+HUVECs group respectively.The proliferation capacity of hDPSCs of each group was examined by CCK-8 and EdU on day 7.Immunofluorescence detected the expression of Stro-1,and Real-time PCR was used to detect the expression of Oct4 and Nanog.Results ①Fluorescence microscopy showed that the trans-fection efficiency was about 80％.Real-time PCR and Western blot results showed that sh-ITGA6 lentivirus effectively interfered with ITGA6 expression in hDPSCs.②CCK-8 results showed that on day 5 of co-culture,the proliferation ability of the sh-ITGA6+HUVECs group was superior to that of the sh-ITGA6 group(P＜0.05);on day 7,the proliferation ability of the sh-NC+HUVECs and sh-ITGA6+HUVECs group was superior to that of the sh-NC and sh-ITGA6 group(P＜0.05).EdU results showed that the DNA synthesis ability of hDPSCs in the co-culture group was significantly stronger than that in the single-culture group(P＜0.05).③Immunofluorescence stai-ning revealed that the expression of Stro-1 in the co-culture group was significantly stronger than that in the single-culture group.④Re-al-time PCR results showed that the expression of Oct4 in the co-culture group was higher than that in the single-culture group(P＜0.05);the expression of Nanog in hDPSCs with sh-ITGA6 was elevated by the addition of HUVECs co-culture(P＜0.05).Conclusion HUVECs significantly enhance the proliferation and stemness of hDPSCs silencing integrin α6.

Purpose To investigate the predictive value of the methylation regulator heterogeneous nuclear ribonu-cleoprotein C(HNRNPC)in the extracellular vesicles(EVs)derived from cervical cancer cells for tumor invasion and metastasis.Methods In vitro experiments were conducted using a human normal cervical epithelial cell line(Hcer-Epic),three cervical cancer cell lines(HeLa,SiHa and CaSki)and human umbilical vein endothelial cells(HU-VECs).HNRNPC protein expression was assessed by Western blot.CaSki cells were transfected with either sh-NC or sh-HNRNPC,and EVs were subsequently isolated from culture supernatant.The effects of EVs on the proliferation,migration and invasion of SiHa cells,as well as on angiogenesis in HUVECs,were investigated respectively.For tissue microarray analysis,normal cervical tissues(n=8),low-grade squamous intraepithelial lesions(LSIL,n=32)and high-grade squamous intraepithelial lesions(HSIL,n=37),and cervical carcinoma specimens(n=153)were ob-tained from our institution.Based on HNRNPC immunoscores,cervical cancer patients were divided into two groups:high(n=99)and low(n=54)HNRNPC expression group,and their clinicopathological features and prognosis were compared.Results Compared with HcerEpic cells,the expression of HNRNPC increased in SiHa,HeLa and CaSki cells.In SiHa cells treated with EVs from CaSki cell,the group receiving EVs from sh-HNRNPC-transfected CaSki cells showed significantly reduced proliferation,colony formation,migration,and invasion compared with the sh-NC-EVs group(P＜0.05).Similarly,in HUVECs treated with CaSki-EVs,compared with sh-NC-EVs group,the sh-HNRNPC-EVs group demonsteated significantly lower protein expression of PCNA and VEGFA and reduced angiogene-sis length(P＜0.05).HNRNPC expression gradually increased during the transformation of cervical epithelial cells(F=106.9,P＜0.001),and was significantlyelevated in HSIL and cervical cancer tissues compared with normal tis-sues(P＜0.001).Moreover,in cervical cancer,high HNRNPC expression was significantly related to poorer cellular differentiation,larger tumor size,parametrial and vaginal infiltration,advanced FIGO stage,and pelvic lymph node metastasis(P＜0.05).Kaplan-Meier analysis showed that that patients with high HNRNPC expression had significant-ly shorter overall survival post-surgery compared with those with low expression(P＜0.05).Conclusion EVs contai-ning HNRNPC contribute to cervical cancer progression by promoting tumor growth,invasion,and angiogenesis,there-by accelerating metastasis.In addition,high HNRNPC expression in cervical cancer tissue is related to poor prognosis of patients,indicating that EV-associated HNRNPC may serve as a potential therapeutic target.

Purpose To investigate the predictive value of the methylation regulator heterogeneous nuclear ribonu-cleoprotein C(HNRNPC)in the extracellular vesicles(EVs)derived from cervical cancer cells for tumor invasion and metastasis.Methods In vitro experiments were conducted using a human normal cervical epithelial cell line(Hcer-Epic),three cervical cancer cell lines(HeLa,SiHa and CaSki)and human umbilical vein endothelial cells(HU-VECs).HNRNPC protein expression was assessed by Western blot.CaSki cells were transfected with either sh-NC or sh-HNRNPC,and EVs were subsequently isolated from culture supernatant.The effects of EVs on the proliferation,migration and invasion of SiHa cells,as well as on angiogenesis in HUVECs,were investigated respectively.For tissue microarray analysis,normal cervical tissues(n=8),low-grade squamous intraepithelial lesions(LSIL,n=32)and high-grade squamous intraepithelial lesions(HSIL,n=37),and cervical carcinoma specimens(n=153)were ob-tained from our institution.Based on HNRNPC immunoscores,cervical cancer patients were divided into two groups:high(n=99)and low(n=54)HNRNPC expression group,and their clinicopathological features and prognosis were compared.Results Compared with HcerEpic cells,the expression of HNRNPC increased in SiHa,HeLa and CaSki cells.In SiHa cells treated with EVs from CaSki cell,the group receiving EVs from sh-HNRNPC-transfected CaSki cells showed significantly reduced proliferation,colony formation,migration,and invasion compared with the sh-NC-EVs group(P＜0.05).Similarly,in HUVECs treated with CaSki-EVs,compared with sh-NC-EVs group,the sh-HNRNPC-EVs group demonsteated significantly lower protein expression of PCNA and VEGFA and reduced angiogene-sis length(P＜0.05).HNRNPC expression gradually increased during the transformation of cervical epithelial cells(F=106.9,P＜0.001),and was significantlyelevated in HSIL and cervical cancer tissues compared with normal tis-sues(P＜0.001).Moreover,in cervical cancer,high HNRNPC expression was significantly related to poorer cellular differentiation,larger tumor size,parametrial and vaginal infiltration,advanced FIGO stage,and pelvic lymph node metastasis(P＜0.05).Kaplan-Meier analysis showed that that patients with high HNRNPC expression had significant-ly shorter overall survival post-surgery compared with those with low expression(P＜0.05).Conclusion EVs contai-ning HNRNPC contribute to cervical cancer progression by promoting tumor growth,invasion,and angiogenesis,there-by accelerating metastasis.In addition,high HNRNPC expression in cervical cancer tissue is related to poor prognosis of patients,indicating that EV-associated HNRNPC may serve as a potential therapeutic target.

Objective To assess the clinical efficacy and safety of rectal mucosal columnar suturing(Block technique)in combination with lauromacrogol injection for the treatment of female rectocele.Methods This retro-spective study analyzed 90 female patients with rectocele who were treated at Sir Run Run Hospital,Nanjing Medical University,from January 2022 to December 2023.Patients were categorized into three groups according to their surgical treatments:Block combined with lauromacrogol injection(BP group,n=30),Block alone(B group,n=30),and Procedure for Prolapse and Hemorrhoids(PPH,H group,n=30).The study compared general clinical data,perioperative indicators,Longo′s Obstructed Defecation Syndrome(Longo′s ODS)scores,the degree of recto-cele before and after surgery,anorectal manometry parameters,surgical efficacy,and perioperative complications among the three groups.Results The intraoperative blood loss in Group H was significantly higher compared to Groups B and BP(P<0.05).In terms of the 24-hour postoperative VAS score and hospital stay duration,Group BP demonstrated superior outcomes relative to Groups H and B(P<0.05).Preoperatively,there were no significant differences among the three groups regarding Longo′s ODS score,rectocele depth,resting anal pressure,or residual anal pressure(P>0.05).Postoperatively,Group BP exhibited significantly better Longo′s ODS scores and rectocele depth compared to Groups B and H(P<0.05).Although no significant differences were observed in postoperative resting and residual anal pressures among the three groups(P>0.05),the values in Group BP were closer to the normal range.The overall efficacy rate in Group BP was 93.3%,which was higher than the 73.3%in Group B and 66.7%in Group H(P<0.05).There was no significant difference in the complication rate across the three groups(P>0.05).Conclusions Block combined with lauromacrogol injection is a safe and effective treatment for female rectocele,demonstrating superior efficacy compared to both PPH and Block alone.This method not only effectively restores the physiological anatomy of the female rectum but also significantly improves clinical symptoms.

Objective To assess the clinical efficacy and safety of rectal mucosal columnar suturing(Block technique)in combination with lauromacrogol injection for the treatment of female rectocele.Methods This retro-spective study analyzed 90 female patients with rectocele who were treated at Sir Run Run Hospital,Nanjing Medical University,from January 2022 to December 2023.Patients were categorized into three groups according to their surgical treatments:Block combined with lauromacrogol injection(BP group,n=30),Block alone(B group,n=30),and Procedure for Prolapse and Hemorrhoids(PPH,H group,n=30).The study compared general clinical data,perioperative indicators,Longo′s Obstructed Defecation Syndrome(Longo′s ODS)scores,the degree of recto-cele before and after surgery,anorectal manometry parameters,surgical efficacy,and perioperative complications among the three groups.Results The intraoperative blood loss in Group H was significantly higher compared to Groups B and BP(P<0.05).In terms of the 24-hour postoperative VAS score and hospital stay duration,Group BP demonstrated superior outcomes relative to Groups H and B(P<0.05).Preoperatively,there were no significant differences among the three groups regarding Longo′s ODS score,rectocele depth,resting anal pressure,or residual anal pressure(P>0.05).Postoperatively,Group BP exhibited significantly better Longo′s ODS scores and rectocele depth compared to Groups B and H(P<0.05).Although no significant differences were observed in postoperative resting and residual anal pressures among the three groups(P>0.05),the values in Group BP were closer to the normal range.The overall efficacy rate in Group BP was 93.3%,which was higher than the 73.3%in Group B and 66.7%in Group H(P<0.05).There was no significant difference in the complication rate across the three groups(P>0.05).Conclusions Block combined with lauromacrogol injection is a safe and effective treatment for female rectocele,demonstrating superior efficacy compared to both PPH and Block alone.This method not only effectively restores the physiological anatomy of the female rectum but also significantly improves clinical symptoms.

Objective To investigate the effect of human umbilical vein endothelial cells(HUVECs)on the proliferation and stem-ness of human dental pulp stem cells(hDPSCs)silencing with integrin α6(ITGA6).Methods ITGA6 silencing lentivirus was used to interfere the ITGA6 expression of hDPSCs,and its silencing efficiency was detected.Then HUVECs were added to the chambers to co-culture,and the experiments were divided into four groups(sh-NC,sh-ITGA6,sh-NC+HUVECs and sh-ITGA6+HUVECs).hDP-SCs in the sh-NC and sh-ITGA6 groups were transfected with sh-NC and sh-ITGA6 respectively.hDPSCs transfected with sh-NC and sh-ITGA6 were co-cultured with HUVECs in the sh-NC+HUVECs group and sh-ITGA6+HUVECs group respectively.The proliferation capacity of hDPSCs of each group was examined by CCK-8 and EdU on day 7.Immunofluorescence detected the expression of Stro-1,and Real-time PCR was used to detect the expression of Oct4 and Nanog.Results ①Fluorescence microscopy showed that the trans-fection efficiency was about 80％.Real-time PCR and Western blot results showed that sh-ITGA6 lentivirus effectively interfered with ITGA6 expression in hDPSCs.②CCK-8 results showed that on day 5 of co-culture,the proliferation ability of the sh-ITGA6+HUVECs group was superior to that of the sh-ITGA6 group(P＜0.05);on day 7,the proliferation ability of the sh-NC+HUVECs and sh-ITGA6+HUVECs group was superior to that of the sh-NC and sh-ITGA6 group(P＜0.05).EdU results showed that the DNA synthesis ability of hDPSCs in the co-culture group was significantly stronger than that in the single-culture group(P＜0.05).③Immunofluorescence stai-ning revealed that the expression of Stro-1 in the co-culture group was significantly stronger than that in the single-culture group.④Re-al-time PCR results showed that the expression of Oct4 in the co-culture group was higher than that in the single-culture group(P＜0.05);the expression of Nanog in hDPSCs with sh-ITGA6 was elevated by the addition of HUVECs co-culture(P＜0.05).Conclusion HUVECs significantly enhance the proliferation and stemness of hDPSCs silencing integrin α6.