Objective:To discuss the expression of Rh family C glycoprotein(RHCG)in the esophageal squamous cell carcinoma(ESCC)tissue and its effect on the malignant biological behavior of ESCC cells,and to clarify the value of RHCG as a diagnostic and prognostic marker for the ESCC patients.Methods:A total of 143 ESCC tissue samples and 105 adjacent normal tissue samples were collected.Using immunohistochemical staining method,141 ESCC samples were divided into two groups:RHCG low expression group(immunohistochemistry score≤6)and RHCG high expression group(immunohistochemistry score>6).Immunohistochemical method was used to detect the RHCG protein expression in 143 ESCC tissues and 105 normal tissues,and the relationship between the clinicopathological characteristics of the ESCC patients was analyzed.Receiver operating characteristic(ROC)curve and Kaplan-Meier survival analysis were used to evaluate the value of RHCG in diagnosis and prognosis of the ESCC patients;univariate and multivariate COX regression analysis were used to determine the independent risk factors affecting the prognosis of the ESCC patients.Gene Expression Profiling Interactive Analysis(GEPIA2)database was used to analyze the expression of RHCG mRNA in various tumor tissues.The ESCC TE-1 cells were cultured and transfected in to 6-well cell culture plates with different Lipofectamine2000∶RHCG ratios;the cells in RHCG transfection group were transfected with weights of 2.0,2.5,and 3.0 μg for 24 and 48 h,respectively,and the cells in NC group transfected with empty vector as control.Western blotting method was used to detect the RHCG protein expression level in the TE-1 cells in various groups after transfection at different concentrations and verify the optimal transfection conditions;cell counting kit-8(CCK-8)assay was used to detect the proliferation activities of the TE-1 cells;plate clone formation assay was used to detect the colony formation numbers of the TE-1 cells;Transwell chamber assay was used to detect the numbers of migrating TE-1 cells.Results:Compared with adjacent normal tissue,the RHCG gene expression level in various cancer tissues including ESCC,glioblastoma multiforme,and head and neck squamous cell carcinoma was significantly decreased(P<0.05).RHCG protein was mainly located on the cell membrane of normal esophageal squamous epithelial cells;the RHCG protein expression intensity in ESCC tissues was lower than that in adjacent normal esophageal tissue(χ2=109.373,P<0.001),and the patients in RHCG low expression group had poorer differentiation than those in RHCG high expression group(P=0.041).The area under the curve(AUC)value of RHCG for diagnosing ESCC was 0.86,with sensitivity and specificity of 95.1%and 75.0%,respectively;the Kaplan-Meier survival analysis results showed that compared with high RHCG expression group,the patients in low RHCG expression group had shorter survival time and poorer prognosis[harard ratio(HR)=0.269,95%confidence interval(CI):0.113-0.639,P=0.020];the COX regression analysis results showed that low RHCG expression could serve as an independent risk factor affecting the prognosis of ESCC[HR=4.569,95%CI=1.315-15.877,P=0.017)].The Western blotting results verified that the optimal transfection condition was 3.0 μg RHCG plasmid for 48 h,at which time RHCG overexpression was optimal and RHCG protein expression level was highest.The CCK-8 assay results showed that compared with control group,the proliferation activity in RHCG overexpression group was decreased on the 4th day after cell seeding(P<0.001).In the TE-1 cells,the colony formation number of the TE-1 cells in RHCG over-expression group was lower than that in control group(t=17.70,P<0.001).The Transwell chamber assay results showed that compared with control group,the number of migrating cells in RHCG over-expression group was decreased(t=23.74,P<0.001).Conclusion:RHCG expression is decreased in ESCC tissues and associated with poor prognosis in ESCC patients;overexpression of RHCG can inhibit the proliferation and migration of the TE-1 cells,providing a theoretical basis for RHCG as a novel diagnostic and prognostic marker and therapeutic target for ESCC.

Background and purpose:BRCA1/2 plays an important role in maintaining the genome stability.Whether BRCA1/2 germline mutation could increase the tumor sensitivity to radiotherapy,thereby inducing secondary primary cancer after radiotherapy is unclear.This study aimed to investigate whether postoperative radiotherapy is a risk factor for the development of second primary cancer in triple-negative breast cancer(TNBC)patients with BRCA1/2 germline mutation.Methods:This research was based on a previously reported retrospective cohort,i.e.,the Fudan University Shanghai Cancer Center TNBC cohort.Between January 1,2007 and December 31,2014,a total of 292 female TNBC patients with BRCA1/2 mutation were enrolled.We performed logistic regression analysis in patients without BRCA1/2 germline mutation(n=261)and BRCA1/2 germline mutation patients(n=31),respectively,to assess the risk factors affecting the incidence of second primary cancer.We then performed interactive analysis on the above two analyses to evaluate the interactive effect between BRCA1/2 germline mutation and postoperative radiotherapy.P<0.05 indicates a statistically significant difference.The research was approved by Fudan University Shanghai Cancer Center TNBC Ethics Committee(050432-4-2108),and each patient provided written informed consent.Results:Logistic regression analysis in patients with BRCA1/2 germline mutations showed that postoperative radiotherapy significantly increased the risk of secondary primary disease compared to non-radiotherapy[odds ratio(OR)=2.475,95%confidence interval(CI):1.933-3.167,P<0.001].In patients without BRCA1/2 germline mutation,the effect of radiotherapy on the incidence of second primary tumor was not significant.There was a significant interaction between BRCA1/2 germline mutation and postoperative radiotherapy for the incidence of secondary primary cancer(OR=9.710,95%CI:0.320-295.250,P=0.193).Conclusion:Although statistical analysis results show that patients with BRCA1/2 germline mutations have an increased risk of developing a second primary tumor after postoperative radiotherapy compared to patients who have not received radiotherapy,there is no significant correlation between BRCA1/2 germline mutations and radiotherapy for the development of a second primary tumor.Therefore,patients with BRCA1/2 germline mutations who receive radiotherapy after surgery may not increase the risk of developing a second primary tumor.

Objective To explore the role and efficacy of VEGF and HGF gene adenovirus vector in promoting angiogenesis in ischemic tissue.Methods 84 Kunming mice were randomly divided into sham group,control group,VEGF group,HGF group and VEGF+HGF group,and the left lower limb ischemia model was established.The blood supply of ischemic tissue was observed by rheometer,and the expression levels of VEGF and HGF in each group were detected by Western Blot and ELISA.Immunohistochemical staining was used to detect angiogenesis(CD31,SMA)in ischemic tissues.Safety was assessed by side effects during treatment in mice.Results After the successful modeling,the blood flow velocity of the left lower limb in each group decreased significantly.On the 7th day after operation,the blood flow of the left lower limb in each group was significantly better than that on the 0th day after operation(P<0.05),and the blood flow of the left lower limb in Ad-VEGF-HGF group was significantly better than that in other groups(P<0.05).On the 28th day after operation,the blood flow of the left lower limb in Ad-VEGF-HGF group gradually stabilized,the blood flow in Ad-VEGF-HGF group was significantly better than that in other groups,and both VEGF group and HGF group were significantly better than the control group(P<0.05).On the 7th,14th,and 28th days following surgery,HGF and VEGF protein levels in the Ad-HGF,Ad-VEGF,and Ad-VEGF-HGF groups were substantially greater than those in the control group(P<0.05).The expression level in the Ad-VEGF-HGF group peaked on the 14th day(all P<0.001)and subsequently declined to preoperative levels on the 28th day after operation.Conclusion Ad-VEGF-HGF gene injection can effectively boost VEGF and HGF protein expression and rapidly reach the relative peak level,encour-aging angiogenesis after lower limb ischemia,increasing blood flow,and improving lower limb circulation.

Objective To explore the clinical characteristics of drug-induced liver injury(DILI),and provide reference for early clinical recognition,diagnosis and treatment of DILI.Methods Clinical datas of 65 patients with DILI hospitalized in the First Affiliated Hospital of Guangxi University of Chinese Medicine from January 2019 to December 2023 were retrospectively analyzed,and their general conditions,using drugs,distribution type of liver injury,traditional Chinese medicine treatment and outcome were statistically analyzed.Results A total of 65 patients were included,including 27 males and 38 females,with an average age of(46.40±19.19)years old.The majority of patients were 41 to 60 years old.Chinese herbal medicine and Chinese patent medicine accounted for the highest proportion of drugs inducing DILI(30.8％).Hepatocyte injury was the most common type of DILI,with 39 cases.The main clinical manifestations of DILI patients were fatigue,abdominal discomfort,aversion to oil,yellow eyes,yellow urine and so on.In the treatment of traditional Chinese medicine,liquorice root was the most frequently used,with a frequency of 60.7％.After active treatment,59 patients were cured and discharged from hospital.Conclusion Hepatocyte injury is the main type of DILI.Clinical monitoring of hepatotoxicity of traditional Chinese herbal medicine and Chinese patent medicine should be strengthened to reduce the damage to liver caused by adverse drug reactions.

Atrial fibrillation(AF),a prevalent clinical tachyarrhythmia,has seen an increase in incidence with advancing age over recent decades.AF leads to severe complications,including stroke and heart failure,and is a sig-nificant cause of mortality.The pathogenesis of AF involves critical factors such as atrial electrical and structural remodeling,oxidative stress,and inflammatory responses.Recent studies highlight the pivotal role of macrophages in the initiation and progression of AF.This study focuses on the contributions of macrophages to the mechanisms of AF and summarizes cur-rent research findings.

Objective To observe the clinical efficacy of autologous hair follicle stem cell transplantation for hair restoration in patients with alopecia. Methods Ten patients with alopecia, including five with androgenic alopecia and five with seborrheic alopecia, were selected from November 2022 to May 2023. They were treated with autologous hair follicle stem cell transplantation. Data before treatment were used as the control group, and data after treatment were used as the study group. Follow-up was conducted for 3 to 8 months. The FPF scoring system was designed to collect data and evaluate the treatment effect. Results Nine patients had an improvement score of ≥7 points, and one patient had an improvement score of 2 points, with a treatment excellence rate of 90% (9/10). Compared with the total score of (2.81±0.92) points in the control group, the total score of the study group was (9.00±2.21) points, and the difference was statistically significant (P < 0.001). Conclusion Autologous hair follicle stem cell transplantation for hair restoration is safe and effective, and has a broad range of indications.

Humans ; Phosphorylation ; Serine/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Signal Transduction ; Neovascularization, Pathologic

Radiocarpal fracture-dislocation (RFD) is a rare injury normally associated with the destruction of bones, joints and ligaments. The improper diagnosis and treatment of RFD will cause severe complications and affect the long-term function of wrist joints. The difficulties of clinical diagnosis and treatment lie in the accurate diagnosis, identification and reconstruction of the structure of specific injury. As the foreign and domestic literatures are mainly case analyses or systemic case reports rather than large-scale reports, there still lacks a systemic knowledge of the standard diagnosis and treatment of RFD clinically, thus leading to problems such as missed diagnosed or misdiagnosed, improper application of treatment methods and incomplete reconstruction. Therefore, the authors reviewed relevant literatures about the features, diagnosis and treatment of RFD, in order to provide references for the clinical diagnosis and treatment of RFD.

Objective:To provide a new morning check method for the output dose stability of the multileaf collimator (MLC) of the CyberKnife M6 (CK-M6) system.Methods:The CT images of a verification phantom with a size of 20 cm × 20 cm × 10 cm were transmitted into the Precision Treatment Plan ning System (ver. 1.1.1.1). The high-precision alignment between the accelerator output front and the fixed position of the phantom surface was achieved using the fiducial tracking method. A 10 cm × 10 cm radiation field was formed by the MLC and a DailyCheck plan with an output of 200 MU was designed. The repeatability, sensitivity, and accuracy of the DailyCheck plan were measured, and the CK-M6 system was continuously tested for one month using the artificial fixed method and the DailyCheck plan designed in this study. Results:The average and the standard deviation of 10 repeated measurements by the DailyCheck plan were 492.28 pC and 0.09, respectively, indicating good stability. There was a linear correlation between the measured values and the output dose, with a correlation coefficient of R2 > 0.999. Moreover, there was a position deviation of 2 mm between the phantom and the accelerator output front, and the result ant effect on the measured values was equivalent to a dose deviation caused by an output of 1.24 MU. The result from the continuous measurement of both the artificial fixed method and the DailyCheck plan fell within permissible limits, showing high consistency. Conclusions:The DailyCheck plan established through the fiducial tracking of a verification phantom can achieve the convenient, quick, and accurate daily detection of the output dose stability of the MLC of CK-M6. Therefore, this method can be widely applied in the clinical quality control of the CK-M6 system.

The CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated) system is an "adaptive immune system" found in the genomes of bacteria and archaea which is mediated by RNA and resists foreign nucleic acid invasion.Take advantage of specific recognition of target nucleic acid, CRISPR-Cas system can efficiently edit their target site or accurately regulate gene expression, and now have been developed into a powerful tool for gene editing.According to the different compositions of the effector complex, the system has been divided into two categories: class 1 (type I, type IV, and type III) and class 2 (type II, type V, and type VI).Class 2 system, like the CRISPR-Cas9, is widely used in basic research due to the earliest discovery and best research.However, class 1 has not been maturely developed and utilized though it makes up 90% of the entire CRISPR-Cas system.In this essay, the classification of subtype, the assembly of Cascade complex, the cleavage and degradation mechanism of Cas3, and the application in gene editing of class 1 type I CRISPR-Cas system will be discussed and summarized to provide new ideas and methods for further mechanism studying and application of this category.