Objective: To identify biomolecular markers closely related to the occurrence of kidney renal clear cell carcinoma (KIRC) and verify their expression levels in clinical samples. Methods: Stage Ⅰ KIRC mRNA sequencing data were obtained from The Cancer Genome Atlas (TCGA).Principal component analysis (PCA) was used for dimensionality reduction to screen differentially expressed genes (DEGs)，which then underwent GO and KEGG analyses.Weighted gene co-expression network analysis (WGCNA) was used to screen genes significantly related to KIRC，and a protein-protein interaction (PPI) network was constructed to screen hub genes.The diagnostic value of hub genes was evaluated with receiver operating characteristic (ROC) curve，and their prognostic value was analyzed using survival curve plots.The correlation between the mRNA expressions of hub genes and the pathological stages of KIRC was analyzed.Clinical samples of 20 patients with stage Ⅰ KIRC treated in our hospital were included，and the expressions of the hub genes in cancerous and adjacent tissues were detected with reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR)，Western blotting，and enzyme-linked immunosorbent assay (ELISA). Results: A total of 8223 DEGs were screened out，including 4092 up-regulated ones and 4131 down-regulated ones.GO analysis showed that DEGs were related to bioadhesion，plasma membrane composition，and transporter activity.KEGG analysis showed that DEGs were related to pathways such as cell adhesion molecules，cytokine-cytokine receptor interactions，and interactions between viral proteins and cytokines and cytokine receptors.WGCNA analysis obtained 171 genes that were significantly related to stage Ⅰ KIRC.The hub gene，lymphocyte cytosolic protein 2 (LCP2)，screened out by the PPI network，was significantly related to stage Ⅰ KIRC.The area under the ROC curve was 0.96.The expression level was negatively correlated with the overall survival rate of patients.The expression of LCP2 was related to the stage and lymph node metastasis.Clinical verification showed that the mRNA and protein relative expressions of LCP2 in KIRC tissues were significantly higher than those in adjacent tissues (P<0.000 1). Conclusion: LCP2 is significantly up-regulated in stage Ⅰ KIRC tissues and can be used as a potential biomarker for the early diagnosis and treatment of KIRC.

ObjectivesTo analyze the spatial and temporal clustering characteristics and related influencing factors of late diagnosis of HIV/AIDS in Lanzhou， to identify its high-risk areas and time trends in Lanzhou， and to provide a theoretical basis for developing targeted HIV/AIDS prevention and control strategies in Lanzhou. MethodsThe subjects of this study were adult HIV/AIDS cases reported in Lanzhou City between 2011 and 2018. Data used in the study were sourced from the Lanzhou Center for Disease Control and Prevention and the Lanzhou Statistical Yearbook. To analyze the spatial distribution characteristics and influencing factors of the relative risk （RR） of late HIV/AIDS diagnosis， Bayes spatial-temporal model was used. ResultsA total of 1984 new HIV/AIDS cases were reported in Lanzhou from 2011 to 2018， with an mean age of 37.51 years and predominantly male （91.8%）. The number of late diagnosis cases was 982， with an mean age of 39.67 years and a predominance of males （91.8%）. Late diagnosis was more common in older individuals and women with HIV/AIDS. Chengguan District （51.1%）， Anning District （50.3%） and Yuzhong County （51.9%） had an above-average proportion of late diagnosis of HIV/AIDS. The proportion of late diagnosis cases in Lanzhou showed a fluctuating upward trend from 2011 to 2018. The results of Bayes spatial-temporal model showed that the risk of late HIV/AIDS diagnosis in Lanzhou had fluctuated from 2011 to 2015， and then increased rapidly after 2015 ［RR （95% credibility interval， 95%CI） increased from 1.01 （0.84， 1.23） to 1.11 （0.77， 1.97）］； the trends of risk of late diagnosis in Honggu district and three counties were similar to the overall trend in Lanzhou city， while the risk of late diagnosis in Chengguan District and Qilihe District showed a decreasing trend. The regions with the RR for late diagnosis greater than 1 included Yongdeng County （RR=1.07， 95% CI： 0.55， 1.96）， Xigu District （RR=1.04， 95% CI： 0.67， 1.49）， Chengguan District （RR=2.41， 95% CI： 0.85， 6.16）， and Qilihe District （RR=2.03， 95% CI： 1.10， 3.27）. Besides， the heatmap analysis showed that Chengguan District and Qilihe District were the hot spots. The influencing factors analysis showed that the higher GDP per capita （RR=0.65， 95% CI： 0.35， 0.90） and the larger proportion of males with HIV/AIDS cases （RR=0.53， 95% CI： 0.19， 0.92） could lead to the lower the relative risk of late HIV/AIDS diagnosis. However， the higher the population density （RR=1.35， 95% CI： 1.01， 1.81） caused the higher the risk of late diagnosis. ConclusionOur study shows the risk of late diagnosis of HIV/AIDS in Lanzhou was on the rise， and there are significant regional differences. GDP per capita， the proportion of males in HIV/AIDS cases and population density are influencing factors in the late diagnosis of HIV/AIDS. Therefore， for regions with a high risk of late diagnosis or related risk factors， targeted HIV screening and prevention services should be given priority in order to reduce the proportion and risk of late diagnosis of HIV/AIDS.

Objective To compare and analyze the differences between customized models and commercial universal models in the segmentation of organs-at-risk in cervical cancer,and to investigate the feasibility of customized models.Methods A retrospective analysis was conducted on 270 cervical cancer patients.Senior clinicians manually delineated organs-at-risk,including the bladder,rectum,small intestine,pelvic bone marrow,femoral heads,and kidneys.The cases were randomly selected to develop customized models,with 202 cases allocated to the training set,38 cases to the test set,and 30 cases to the validation set.The universal and customized models were used for segmentation on the test set,and the automatic segmentation results obtained by the two models were compared with manual segmentation results to assess the performance of the customized model.Results Both customized model and universal model had comparable DSC values to manual segmentation,demonstrating satisfactory delineation outcomes(DSC values ranging from 0.7 to 0.9).However,in terms of deviation of centroid and 95％Hausdorff distance,the customized model surpassed the universal model.Conclusion Compared with the universal model,the customized model offers superior accuracy in delineating the structures of organs-at-risk in cervical cancer.As the customized model is optimized based on specific datasets,it provides precise support for clinical decision-making and holds promising applications in the treatment of cervical cancer.

OBJECTIVE: To carry out prenatal diagnosis and genetic analysis for a fetus with disorders of sex development (DSDs). METHODS: A fetus with DSDs who was identified at the Shenzhen People's Hospital in September 2021 was selected as the study subject. Combined molecular genetic techniques including quantitative fluorescence PCR (QF-PCR), multiplex ligation-dependent probe amplification (MLPA), chromosomal microarray analysis (CMA), quantitative real-time PCR (qPCR), as well as cytogenetic techniques such as karyotyping analysis and fluorescence in situ hybridization (FISH) were applied. Ultrasonography was used to observe the phenotype of sex development. RESULTS: Molecular genetic testing suggested that the fetus had mosaicism of Yq11.222qter deletion and X monosomy. Combined with the result of cytogenetic testing, its karyotype was determined as mos 45,X[34]/46,X,del(Y)(q11.222)[61]/47,X,del(Y)(q11.222),del(Y)(q11.222)[5]. Ultrasound examination suggested hypospadia, which was confirmed after elective abortion. Combined the results of genetic testing and phenotypic analysis, the fetus was ultimately diagnosed with DSDs. CONCLUSION This study has applied a variety of genetic techniques and ultrasonography to diagnose a fetus with DSDs with a complex karyotype.
Prenatal Diagnosis ; Mosaicism ; Chromosomes, Human, X ; Chromosomes, Human, Y ; Humans ; Male

OBJECTIVE: To investigate the role of the SIRT1/NF-κB pathway in mediating the effect of puerarin against lipopolysaccharide (LPS)-induced acute kidney injury (AKI). METHODS: Fifteen BALB/C mice were randomized into control group, LPS group and puerarin treatment group, and in the latter two groups, the mice were given an intraperitoneal injection of LPS (5 mg/kg), followed by daily injection of normal saline for 3 days or injection of puerarin (25 mg/kg) given 1 h later and then on a daily basis for 3 days. On day 5 after modeling, the kidney tissues were taken for histological observation and detection of cell apoptosis. The renal function indexes including urea nitrogen (BUN), serum creatinine (Scr) and kidney injury molecule 1 (KIM-1) and the levels of tumor necrosis factor (TNF-α) and interleukin 1β (IL-1β) were measured, and the expressions of SIRT1 and NF-κB-p65(acetyl K310) in the renal tissues were detected. RESULTS: Intraperitoneal injection of LPS caused obvious glomerular capillary dilatation, hyperemia, renal interstitial edema, and renal tubular epithelial cell swelling and deformation in the mice. The mouse models of LPS-induced AKI also showed significantly increased renal tubular injury score and renal cell apoptosis (P < 0.01) with increased serum levels of BUN, Scr, KIM-1, TNF-α and IL-1β (P < 0.01), enhanced renal expressions of TNF-α, IL-1β and NF-κB p65(acetyl K310) (P < 0.01) and lowered renal expression of SIRT1 (P < 0.05). Treatment with puerarin effectively alleviated LPS-induced renal interstitial edema and renal tubular epithelial cell shedding, lowered renal tubular injury score (P < 0.01) and renal cell apoptosis rate (P < 0.01), and decreased serum levels of BUN, Scr, KIM, TNF-α and IL-1β (P < 0.01). Puerarin treatment significantly reduced TNF-α, IL-1β and NF-κB p65 (acetyl K310) expression in the renal tissue (P < 0.05) and increased SIRT1 expression by 17% (P < 0.05) in the mouse models. CONCLUSION Puerarin can effectively alleviate LPS-induced AKI in mice possibly by modulating the SIRT1/NF-κB signaling pathway.
Animals ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; Lipopolysaccharides ; Sirtuin 1 ; Tumor Necrosis Factor-alpha ; Acute Kidney Injury ; Disease Models, Animal ; Edema

Objective:To investigate the value of hemorrhage after thrombolytic (HAT) score and neutrophil to lymphocyte ratio (NLR) in combination predicting symptomatic intracerebral hemorrhage (sICH) after intravenous thrombolysis in patients with acute ischemic stroke (AIS).Methods:Consective patients with AIS received intravenous thrombolysis with ateplase in Tianjin TEDA Hospital from January 2016 to December 2021 were retrospectively enrolled. sICH was defined as cerebral CT showing hemorrhage at any part of the brain after intravenous thrombolysis, and the National Institutes of Health Stroke Scale (NIHSS) score was increased by≥4 compared with the baseline, or there was a manifestation indicating clinical aggravation. Univariate analysis was used to compare the baseline data of sICH group and non-sICH group. A binary multivariate logistic regression model was used to determine the independent influencing factors of sICH. The receiver operating characteristic (ROC) curve was used to evaluate the value of HAT score and NLR in combination predicting sICH. Results:A total of 429 patients with AIS were enrolled. Univariate analysis showed that there were significant differences in atrial fibrillation, systolic blood pressure, NLR, HAT score and NIHSS score between the sICH group and the non-sICH group (all P<0.05). Multivariate analysis showed that NLR (odds ratio [ OR] 1.405, 95% confidence interval [ CI] 1.193-2.958), HAT score ( OR 1.512, 95% CI 1.207-3.169) and NIHSS score ( OR 1.221, 95% CI 1.082-2.634) had significant independent correlation with sICH after adjusting for atrial fibrillation and systolic blood pressure. The ROC curve showed that the areas under the curve of HAT score, NLR and their combination predicting sICH were 0.719 (95% CI 0.609-0.832), 0.723 (95% CI 0.618-0.835) and 0.854 (95% CI 0.765-0.931), respectively. The areas under the curve of the two methods in combination were significantly larger than those of the single method ( P=0.029 and 0.032, respectively), and their sensitivity and specificity were 74.1% and 83.5% respectively. Conclusion:Combined HAT score and NLR is of high value in predicting sICH after intravenous thrombolysis in patients with AIS, and has clinical application potential.

Objective To explore the efficacy of debridement combined with antibiotic-loaded artificial bone in the treatment of clavicular osteomyelitis.Methods The data of 45 patients with clavicle osteomyelitis admitted to Luoyang Orthopedic Hospital(Henan Orthopedic Hospital)in Henan Province from January 2012 to June 2022 were retrospectively analyzed.They were divided into treatment group(n = 24)treated with debridement combined with antibiotic-loaded artificial bone,and control group(n = 21)treated with debridement.We compared the operation time,the duration of drainage tube placement,wound healing time,white blood cells(WBC),C-reactive protein(CRP),and erythrocyte sedimentation rate(ESR)in the two groups.Preoperative visual analog scores(VAS)and VAS 1 month after operation,and preoperative Constant-Murley score(CMS)and CMS 12 months after operation were observed.Infections,recurrence and complications in the two groups were recorded in the follow-up.Results All the 45 patients completed the surgery successfully and were followed up for 13 to 35 months,with an average of(23.53±5.11)months.The operation time and the duration of drainage tube place-ment of the treatment group were longer than those of the control group(P<0.05),and there was no significant difference in the wound healing time(P>0.05);preoperative and 14-day postoperative WBC,CRP,and ESR,VAS 1 month after operation,and CMS 12 months after operation were all significantly improved in the two groups.The differences were all statistically significant(P<0.05),and the CMS of the treatment group was higher than that of the control group(P<0.05).During the follow-up,there were less cases of infection recurrence in the treatment group when compared with that in the control group(2/24,8.33%vs.5/21,23.81%).There were 2 cases of aseptic exudation in the treatment group.There were 1 case of bone defect in the treatment group and 5 in the control group,all of which had healed after the second-stage iliac implantation,and the rest did not have the complication of aseptic exudation,bone defects and pathologic fracture.Conclusion Debridement and antibiotic-loaded artificial bone can effectively control the infection and preserve the shape and function of the clavicle,with a low recurrence rate of postoperative infection,simple surgical operation,and no serious complications.It is wor-thy of clinical promotion.

Objective To investigate the repairing effect of ursolic acid extracted from Hippophae rhamnoi-des L.on sciatic nerve injury in rats.Methods After preparing a sciatic nerve damage model,thirty 2-month old Wistar rats were randomly divided into ursolic acid and control groups,with 15 rats in each group.Ursolic acid group was given Ursolic acid 150 mg/(kg·d)by gavage,while the control group was given the same dose of normal saline for 6 weeks.After 6 weeks,H&E staining was used to observe the pathological changes of rat sciatic nerves,and SFI and MNCV were calculated;The expression of NGF in sciatic nerve was detected by immunohistochemistry,and the expression of GAP-43 in L4-L6 spinal cord was detected by Western blotting.Results After 6 weeks,compared to the control group,the number of regenerated nerve fibers in the ursolic acid group increased,the arrangement was slightly regular,the capillary structure was abundant,and the proliferation of fibroblasts decreased;SFI and MNCV%were significantly increased(P<0.05),NGF protein positive expression was significantly increased(P<0.05),and the difference was statistically significant(P<0.05).GAP-43 protein expression in sciatic nerve tissue was significantly increased(P<0.05).Conclusions The intervention of ursolic acid extracted from Hippophae rhamnoides L.can improve the function of rats after sciatic nerve injury,promote the increase of regenerated nerve fibers and the repair of myelin sheath in rats with sciatic nerve injury,which may be related to increased NGF and GAP-43 protein expression in rats.

Pheretima,also called"earthworms",is a well-known animal-derived traditional Chinese medicine that is extensively used in over 50 Chinese patent medicines(CPMs)in Chinese Pharmacopoeia(2020 edi-tion).However,its zoological origin is unclear,both in the herbal market and CPMs.In this study,a strategy for integrating in-house annotated protein databases constructed from close evolutionary relationship-sourced RNA sequencing data from public archival resources and various sequencing al-gorithms(restricted search,open search,and de novo)was developed to characterize the phenotype of natural peptides of three major commercial species of Pheretima,including Pheretima aspergillum(PA),Pheretima vulgaris(PV),and Metaphire magna(MM).We identified 10,477 natural peptides in the PA,7,451 in PV,and 5,896 in MM samples.Five specific signature peptides were screened and then validated using synthetic peptides;these demonstrated robust specificity for the authentication of PA,PV,and MM.Finally,all marker peptides were successfully applied to identify the zoological origins of Brain Heart capsules and Xiaohuoluo pills,revealing the inconsistent Pheretima species used in these CPMs.In conclusion,our integrated strategy could be used for the in-depth characterization of natural peptides of other animal-derived traditional Chinese medicines,especially non-model species with poorly annotated protein databases.

Cordyceps sinensis(C.sinensis)is a widely used and highly valuable traditional Chinese medicine.Several dipeptides have been detected in C.sinensis,but current scientific knowledge of its chemical makeup remains limited.In this study,an improved approach that integrates offline two-dimensional liquid chromatography(2D LC)separation,precursor ion list,library screening,and diagnostic ion filtering was established to systematically screen and characterize dipeptides in C.sinensis.Offline 2D LC integrating hydrophilic interaction LC and reverse phase separations was established to eliminate interference and identify the target dipeptides.A library containing the potential 400 dipeptides was created,and a precursor ion list with all theoretical precursor ions was adopted to trigger the MS/MS scan with high sensitivity.To identify dipeptides,the type and connection sequence of amino acids were determined according to the product ions.Ile and Leu residues were differentiated for the first time according to the characteristic ion at m/z 69.07.Ultimately,170 dipeptides were identified or tentatively characterized from C.sinensis,and most are reported for the first time in this species herein.In addition,the identified dipeptides were also applied for discrimination among the three Cordyceps species,and 11 markers were identified.The obtained results provide a deeper understanding of the chemical basis of C.sinensis.