Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder affecting a substantial proportion of women of reproductive age. It is frequently associated with ovulatory dysfunction, infertility, and an increased risk of chronic metabolic diseases. A hallmark pathological feature of PCOS is the arrest of follicular development, closely linked to impaired intercellular communication between the oocyte and surrounding granulosa cells. Transzonal projections (TZPs) are specialized cytoplasmic extensions derived from granulosa cells that penetrate the zona pellucida to establish direct contact with the oocyte. These structures serve as essential conduits for the transfer of metabolites, signaling molecules (e.g., cAMP, cGMP), and regulatory factors (e.g., microRNAs, growth differentiation factors), thereby maintaining meiotic arrest, facilitating metabolic cooperation, and supporting gene expression regulation in the oocyte. The proper formation and maintenance of TZPs depend on the cytoskeletal integrity of granulosa cells and the regulated expression of key connexins, particularly CX37 and CX43. Recent studies have revealed that in PCOS, TZPs exhibit significant structural and functional abnormalities. Contributing factors—such as hyperandrogenism, insulin resistance, oxidative stress, chronic inflammation, and dysregulation of critical signaling pathways (including PI3K/Akt, Wnt/β‑catenin, and MAPK/ERK)—collectively impair TZP integrity and reduce their formation. This disruption in granulosa-oocyte communication compromises oocyte quality and contributes to follicular arrest and anovulation. This review provides a comprehensive overview of TZP biology, including their formation mechanisms, molecular composition, and stage-specific dynamics during folliculogenesis. We highlight the pathological alterations in TZPs observed in PCOS and elucidate how endocrine and metabolic disturbances—particularly androgen excess and hyperinsulinemia—downregulate CX43 expression and impair gap junction function, thereby exacerbating ovarian microenvironmental dysfunction. Furthermore, we explore emerging therapeutic strategies aimed at preserving or restoring TZP integrity. Anti-androgen therapies (e.g., spironolactone, flutamide), insulin sensitizers (e.g., metformin), and GLP-1 receptor agonists (e.g., liraglutide) have shown potential in modulating connexin expression and enhancing granulosa-oocyte communication. In addition, agents such as melatonin, AMPK activators, and GDF9/BMP15 analogs may promote TZP formation and improve oocyte competence. Advanced technologies, including ovarian organoid models and CRISPR-based gene editing, offer promising platforms for studying TZP regulation and developing targeted interventions. In summary, TZPs are indispensable for maintaining follicular homeostasis, and their disruption plays a pivotal role in the pathogenesis of PCOS-related folliculogenesis failure. Targeting TZP integrity represents a promising therapeutic avenue in PCOS management and warrants further mechanistic and translational investigation.

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

Objective:To construct the sphingosine kinase 1(SPHK1)overexpression lentiviral vector,and to establish the SKOV3 lentiviral stable transfection cell line.Methods:According to the SPHK1 data information provided by the National Center for Biotechnology Information(NCBI)database,the primers were designed and synthesized,the target gene was amplified,and connected to the GV492 plasmid treated with Bam HⅠ and AgeⅠ restriction enzymes to construct the SPHK1 overexpression lentiviral vector;the positive clones were selected for PCR and sequencing identification;the lentiviral plasmid and the lentiviral packaging auxiliary plasmid were co-transfected into the HEK-293T cells for packaging and titer determination;according to the measured optimal multiplicity of infection(MOI)of 10,the corresponding lentiviral amounts in various groups were transfected into the SKOV3 cells,and the SKOV3 cells were divided into blank group(without treatment),GV492 control group(GV492 control lentivirus infected SKOV3 cells),and GV492-SPHK1 overexpression group(GV492-SPHK1 overexpression lentivirus infected SKOV3 cells,ov-SPHK1 group);the optimal concentration of 2 mg·L-1 puromycin was used to screen the stably transfected SKOV3 cell line;after 48 h,the medium was changed and replaced with 1 mg·L-1 puromycin for screening for 14 d;the morphology and fluorescence expression of the cells were observed under fluorescence microscope;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of SPHK1 mRNA in the SKOV3 cells in various groups;Western blotting method was used to detect the expression level of SPHK1 protein in the SKOV3 cells in various groups.Results:The PCR sequencing results showed that the gene sequence of the SPHK1 overexpression lentiviral vector was completely consistent with the target sequence,and the SPHK1 overexpression lentiviral vector was successfully constructed;the titer determination results showed that the lentiviral titers in GV492 control group and ov-SPHK1 group were 5×1011 and 8×1011 TU·L?1,respectively;the SKOV3 cells in GV492 control group and ov-SPHK1 group were in good state and showed strong fluorescence expression,suggesting that the SKOV3 stable transfection cell line overexpressing SPHK1 was successfully established;the RT-qPCR results showed that compared with blank group and GV492 control group,the expression level of SPHK1 mRNA in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01);the Western blotting results showed that compared with blank group and GV492 control group,the expression level of SPHK1 protein in the SKOV3 cells in ov-SPHK1 group was significantly increased(P<0.01).Conclusion:The SPHK1 overexpression lentiviral vector is successfully constructed,and the SKOV3 stable transfection cell line is established.

Objective To establish TLC identification method,HPLC fingerprint,chemical pattern recognition,and multi-components quantitative analysis methods of Spatholobus suberectus,and identify the quality of 16 batches of Spatholobus su-berectus from different origins.Methods The TLC method was used to identify Spatholobus suberectus qualitatively.HPLC fin-gerprint of Spatholobus suberectus was established.The quality of Spatholobus suberectus was evaluated by chemical pattern recogni-tion technologies,and the contents of Gallocatechin,protocatechuic acid,catechin,epigallocatechin and fermononetin were deter-mined by HPLC.Results TLC method of Spatholobus suberectus was established.Ten common peaks were identified in 16 bat-ches of Spatholobus suberectus,and 6 components were identified by reference substances.The similarity of fingerprint was 0.769-0.990,indicating good similarity.The samples were divided into 3 groups by cluster analysis.Results of the principal component a-nalysis showed that the top 3 samples in the list of comprehensive scores were S16、S1 1、S14.Marker compounds that cause the quality difference of Spatholobus suberectus were screened out through the orthogonal partial least squares discriminant analysis,which were fermononetin,peaks 7 and epigallocatechin.Gallocatechin,protocatechuic acid,catechin,epigallocatechin,and ferm-ononetin had a good linearity in the concentration range(r＞0.999).The content determination analysis showed that there were sig-nificant differences in the contents of the 5 index components among Spatholobus suberectus from different origins.Conclusion The established TLC and HPLC fingerprints of Spatholobus suberectus were stable and reliable,and the HPLC method for multiple active components method provides scientific support for improving the quality control method of Spatholobus suberectus.

This article summarizesd Professor Li Zhigang's experience in treating limb dysfunction after acute disseminated encephalomyelitis(ADEM)by acupuncture.Prof.Li respected the classical theory of Chinese medicine,which is to seek the root cause of the disease,clarified the disease mechanism of"deficiency of Yangming leads to longitudinal tendons and unfavorable belt veins",and followed the principle of"treating flaccidity syndrome only by taking advantage of Yangming",and combined with the morbidity characteristics of ADEM that occurs in children,emphasized the important influence of regulating the root of innate and acquired in the treatment of the disease.The importance of regulating the essence of congenital and acquired constitution in the treatment of this disease was emphasized,taking into account the importance of regulating the mind and spirit.The main acupoints of Yangming Stomach Meridian,Large Intestine Meridian,Spleen Meridian,and Governor Vessel,together with the five main points of the lower limbs and the main points of water-inducing,all together play the role of tonifying the middle energizer,benefiting the essence and filling in the marrow,and strengthening the muscles and bones.

Objective To establish TLC identification method,HPLC fingerprint,chemical pattern recognition,and multi-components quantitative analysis methods of Spatholobus suberectus,and identify the quality of 16 batches of Spatholobus su-berectus from different origins.Methods The TLC method was used to identify Spatholobus suberectus qualitatively.HPLC fin-gerprint of Spatholobus suberectus was established.The quality of Spatholobus suberectus was evaluated by chemical pattern recogni-tion technologies,and the contents of Gallocatechin,protocatechuic acid,catechin,epigallocatechin and fermononetin were deter-mined by HPLC.Results TLC method of Spatholobus suberectus was established.Ten common peaks were identified in 16 bat-ches of Spatholobus suberectus,and 6 components were identified by reference substances.The similarity of fingerprint was 0.769-0.990,indicating good similarity.The samples were divided into 3 groups by cluster analysis.Results of the principal component a-nalysis showed that the top 3 samples in the list of comprehensive scores were S16、S1 1、S14.Marker compounds that cause the quality difference of Spatholobus suberectus were screened out through the orthogonal partial least squares discriminant analysis,which were fermononetin,peaks 7 and epigallocatechin.Gallocatechin,protocatechuic acid,catechin,epigallocatechin,and ferm-ononetin had a good linearity in the concentration range(r＞0.999).The content determination analysis showed that there were sig-nificant differences in the contents of the 5 index components among Spatholobus suberectus from different origins.Conclusion The established TLC and HPLC fingerprints of Spatholobus suberectus were stable and reliable,and the HPLC method for multiple active components method provides scientific support for improving the quality control method of Spatholobus suberectus.

AIM:To investigate the therapeutic effects and potential mechanism of pachymic acid(PA)on li-popolysaccharide(LPS)-induced acute kidney injury(AKI)in mice.METHODS:(1)Genes related to AKI were screened using the DAVID database.Core genes were identified by intersecting related genes and analyzed using Cyto-scape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed through the DAVID database for the cross-targets.Molecular docking and activity assays were conducted on the primary core targets.(2)A total of 100 C57BL/6J mice were randomly divided into five groups:normal control(NC),model(LPS),solvent control(LPS+DMSO),and treatment groups(LPS+PA-10 and LPS+PA-20),with 20 mice in each group.The LPS-AKI model was established by intraperitoneal injection of 18 mg/kg LPS.The treatment groups received 10 mg/kg and 20 mg/kg PA,respectively,and the solvent control group was administered an equivalent dose of DMSO.Mice were euthanized 24 h after injection.Serum was collected for biochemical analysis,and Western blot was used to detect neutro-phil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),caspase-3,cleaved caspase-3,interleu-kin-1β(IL-1β),and monocyte chemoattractant protein-1(MCP-1)protein expression.RT-qPCR was employed to detect inflammatory factor mRNA levels.Molecular docking was used to simulate the optimal binding site of PA to caspase-3.En-zyme activity assays were performed to assess caspase protein activity,and renal lesions were observed via hematoxylin and eosin(HE)staining.Apoptosis was detected by TUNEL staining.RESULTS:(1)Thirty-one potential targets of PA against AKI were identified through network pharmacology.GO and KEGG enrichment analyses indicated that these tar-gets were primarily involved in immune response,inflammatory processes,apoptosis and survival,angiogenesis and hemo-dynamics,oxidative stress,and endoplasmic reticulum stress.Key targets included CASP3(caspase-3),PTGS2,BCL2,CCL2,and CYP219.(2)PA treatment improved renal function and reduced tubular epithelial injury.It significantly de-creased NGAL,KIM-1,and cleaved caspase-3 protein levels,as well as inflammatory factors TNF-α,IL-1β,and MCP-1 mRNA and protein expression.PA also reduced apoptosis of renal tubular epithelial cells.Enzyme activity assays and mo-lecular docking revealed that PA exerted its anti-apoptotic effect by directly binding to caspase-3,thereby inhibiting its ac-tivation by caspase-8.CONCLUSION:PA demonstrated a therapeutic effect in LPS-AKI,potentially through the inhibi-tion of inflammatory factor synthesis and release,as well as the inhibition of caspase-3 activation by caspase-8,reducing apoptosis in renal tubular epithelial cells.

This consensus was developed by the Orthodontic Society of the Chinese Stomatological Association to provide a systematic, scientific, and practical guideline for informed consent in orthodontic care. Orthodontic treatment is typically lengthy, highly individualized, and involves multiple factors such as growth and development, occlusal function, and facial esthetics. Rapid technological advances and diverse risk profiles make the traditional reliance on orthodontist experience or institutional templates insufficient to ensure patients′ full understanding and autonomous decision-making. To address this, the expert panel conducted extensive reviews of domestic and international guidelines, analyzed representative dispute cases, and performed multicenter patient-clinician surveys. Using a multi-round Delphi method, the group established a standardized informed consent framework covering the initial consultation, treatment, and retention phases. The consensus emphasizes that informed consent is not only a fundamental legal and ethical requirement but also a key step in building trust, improving patient compliance, and enhancing treatment satisfaction. Orthodontists should clearly and comprehensively explain treatment plans, potential risks, uncertainties, and associated costs, while respecting the autonomy of patients or guardians, and maintain continuous communication and dynamic evaluation throughout the treatment process. The release of this consensus provides unified and authoritative guidance for clinical orthodontics, helping to standardize informed consent, enhance its transparency, safeguard patient rights, reduce medical risks, and promote high-quality, sustainable development of orthodontic practice.

Objective:To analyze the clinical features and risk factors for renal injury in children with antineutrophil cytoplasmic antibody (ANCA)-positive IgA vasculitis (IgAV).Methods:A case-control study was conducted. Seventy-two ANCA-positive IgAV children hospitalized at the Children′s Hospital of Chongqing Medical University from January 2017 to October 2022 were enrolled as the ANCA-positive group. Propensity score matching (1∶4) using the nearest neighbor was performed with age and gender as covariate, and 288 cases ANCA-negative IgAV children were included as the ANCA-negative group. Patients with renal injury were named ANCA-positive IgAV nephritis (IgAVN) group and ANCA-negative IgAVN group, respectively. The ANCA-positive IgAVN group was further divided into myeloperoxidase (MPO) group and proteinase 3 (PR3) group based on the type of ANCA. Clinical data including manifestations, laboratory tests, renal injury, and prognosis were collected. Comparisons between groups were performed using independent sample t-tests, Mann-Whitney U tests, χ2 tests, or Fisher′s exact tests. Kaplan-Meier curves were used to assess differences in the time to renal injury onset, and multivariate logistic regression was performed to identify independent risk factors for renal injury. Results:Among the 72 ANCA-positive IgAV children (41 males, 31 females, age of 7.7 (5.3, 11.2) years), no significant difference in age or gender was observed compared to the ANCA-negative group (both P>0.05). The ANCA-positive group had higher IgM levels, a higher incidence of recurrent rash, and shorter thrombin time (all P<0.05). Among children with renal injury, the ANCA-positive group showed significant differences in the incidence of hematuria, clinical classification, and grade A prognosis compared to the ANCA-negative group (all P<0.05), but no difference was found in the time to renal involvement onest or renal pathology (all P>0.05). The MPO group had higher rates of microscopic hematuria, gross hematuria, acute renal insufficiency, glomerular sclerosis, and grade B prognosis compared to the ANCA-negative IgAVN group (all P<0.05), with a later onset of renal involvement ( P<0.05). Elevated serum creatinine ( OR=1.08, 95% CI 1.03-1.14) and shortened thrombin time ( OR=0.71, 95% CI 0.55-0.92) were independent risk factors for renal injury in ANCA-positive IgAV children (all P<0.05). Conclusions:Children with ANCA-positive IgAV are more likely to experience recurrent rash. MPO-ANCA-positive IgAVN children have higher risks of hematuria, acute kidney injury and glomerular sclerosis, with later-onset but poorer renal prognosis compared to ANCA-negative IgAVN children. Higher serum creatinine levels and shorter thrombin time may be associated with renal injury in children with ANCA-positive IgAV.

Objective:To explore the therapeutic effect of Honghe Fujie lotion combined with Lactobacillus vaginal capsule on senile vaginitis and its influence on vaginal microenvironment and inflammatory factors in vaginal lavage fluid.Methods:From February 2020 to March 2024 122 patients with senile vaginitis in.first department of gynecology of Northwest Women's and Children's Hospital were retrospectively selected as the study subjects. Among them, 60 patients who adopted Lactobacillus vaginal capsule were included in control group, and 62 patients who received Lactobacillus vaginal capsule combined with Honghe Fujie lotion were enrolled as combined group. The clinical efficacy of the two groups of patients was observed, and the changes of vaginal microenvironment and vaginal lavage fluid inflammatory factors before and after treatment and occurrence of adverse reactions during treatment were compared. The t-test was used for comparing between groups of metric data that conform to normal distribution, the χ2 test was used for comparing between groups of count data, and the rank sum test was used for comparing between groups of rank data. Results:The combined group (95.16%, 59/62) had higher total effective rate after the end of treatment than the control group (85.33%, 50/60) ( χ2=4.48, P=0.034). The levels of tumor necrosis factor- α ((106.22±4.83) ng/L, (109.34±5.52) ng/L), interleukin-6 ((4.19±1.14) ng/L, (4.98±1.42) ng/L) and Toll-like receptor-2 ((5.45±1.02) ng/L, (6.27±1.63) ng/L), vaginal pH value (PH≤4.5: 48 cases, 36 cases) and flora diversity (level 1,2,3,4:51,5,4,2 cases and 32,17,7,4 cases) in combined group and control group after the end of treatment were lower compared with before treatment ((145.27±6.01, 146.19±6.52) ng/L, (13.54±2.21,13.47±2.35) ng/L, (10.21±2.14, 10.34±2.11), (PH≤4.5: 0,0 case, flora diversity level 1,2,3,4:9,26,15,12 cases and 8,24,16,12 cases), the differences were statistically significant (statistical values were t=39.88, t=33.41, t=29.61, t=23.59, t=15.81, t=11.82, χ2=78.32, χ2=51.43, Z=48.77, and Z=21.07, respectively; all P<0.001), and the above-mentioned indicators were lower in combined group , the differences were statistically significant (statistical value s were t=3.33, t=3.39, t=3.34, χ2=4.31, and χ2=10.27, respectively; P value s were 0.001, 0.001, 0.001, 0.038, and 0.001, respectively). There was no statistically significant difference in the vaginal microbiota density between the two groups of patients after treatment compared to before treatment ( χ2=1.01, P=0.314). There was no statistically significant difference in the total incidence of adverse reactions between the two groups of patients during treatment ( χ2=0.18, P=0.675). Conclusion:The combination of Lactobacillus vaginal capsule and Honghe Fujie lotion has an exact efficacy in treating senile vaginitis. It can help to relieve the body's inflammatory response and promote the recovery of normal vaginal microenvironment of patients, and it has good safety. It is worthy of clinical promotion and application.