This article introduced three proved cases of gynecological disease: amenorrhea, climacteric syndrome and breast nodules, which were treated by HE Sai-ping doctor. He insisted that the gynecological diseases could be treated based on spleen and liver, of which regulating qi-blood was the therapeutic core.

Objective To compare the effect of conservative and operative treatment for cervical spinal cord injury without fracture and dislocation (CSCIWFD)and to detect mechanism of injury as well as its relationship to outcome.Methods A retrospective review was conducted on 688 patients with CSCIWFD treated from August 1994 to March 2013.There were 155 patients managed conservatively (conservation group) and 533 surgically (operation group).Neurological function improvement was compared between two groups to detect the correlation of patents' age and treatment methods with outcome.Results The patients were followed up for mean 17.9 months (range,3-36 months).Neurological function was estimated using Japanese Orthopedic Association (JOA) score:(1) the recovery rate of patients aged over 40 years in operation group was better than that in conservation group (P ＜0.05) ; (2) the recovery rate in patients aged under 39 years was unsatisfactory in both groups,with insignificant difference between the two groups (P ＞ 0.05).Conclusions Different age of patients with CSCIWFD has different injury mechanism,injury severity and outcome.Surgery provides better results than conservative treatment for patients aged over 40 years,but both results are poor for patients aged under 39 vears.

Objective To investigate whether resting state-fMRI (RS-fMRI) based on local consistency (ReHo), am?plitude low-frequency fluctuate (ALFF) and fALFF can add meaningful information on preoperative localization of epilepto?genic zone in patients with malformations of cortical development (MCD). Methods Ten epilepsy patients with MCD were studied with RS-fMRI using a 3.0 T scanner. The resting state data were preprocessed and analyzed using SPM8 and REST to generate the activation map. Abnormal ReHo, ALFF and fALFF related blood oxygen level dependent (BOLD) signal changes were compared to video EEG (VEEG),PET,MRI findings and the final result of a comprehensive evaluation-de?fined epileptogenic zone. For operated patients, postoperative resection and histology were compared to BOLD responses. Re?sults The results of spike localization of RS-fMRI were consistent with VEEG, PET, MRI findings and final comprehensive evaluation-defined epileptogenic zone in 6, 8, 6, 7 of 10 investigations. Six operated patients (including two negative results of MRI examination) revealed local abnormal changes but not visible on structural MRI, which was confirmed cortical malfor?mations by pathology after operation (2 heterotopia and 4 cortical dysplasia). Conclusion RS-fMRI may help to delineate the epileptic focus in epilepsy patients with MCD.

OBJECTIVETo investigate the suitable transfection condition of human epidermal cells (hECs) with human epidermal growth factor (EGF) gene by adenovirus vector (Ad-hEGF) and its effects on the biological characteristics of hECs.

METHODShECs were isolated from deprecated human fresh prepuce tissue of circumcision by enzyme digestion method and then sub-cultured. hECs of the third passage were used in the following experiments. (1) Cells were divided into non-transfection group and 5, 20, 50, 100, 150, and 200 fold transfection groups according to the random number table (the same grouping method below), with 3 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in the latter six groups were transfected with Ad-hEGF gene in multiplicities of infection (MOI) of 5, 20, 50, 100, 150, and 200 respectively. The morphology of the cells was observed with inverted phase contrast microscope, and expression of green fluorescent protein of the cells was observed with inverted fluorescence microscope at transfection hour (TH) 24, 48, and 72. (2) Another three batches of cells were collected, grouped, and treated as above, respectively. Then the transfection rate of Ad-hEGF gene was detected by flow cytometer (n=3), the mass concentration of EGF in culture supernatant of cells was detected by enzyme-linked immunosorbent assay (n=6), and the proliferation activity of cells was detected by cell counting kit 8 (CCK8) and microplate reader (n=6) at TH 24, 48, and 72, respectively. (3) Cells were collected and divided into non-transfection group and transfection group, with 6 wells in each group. Cells in non-transfection group were cultured with culture supernatant of cells without transfection, while cells in transfection group were cultured with culture supernatant of cells which were transfected with Ad-hEGF gene in the optimum MOI (50). CCK8 and microplate reader were used to measure the biological activity of EGF secreted by cells on culture day 1, 3, and 5. (4) Cells were collected and divided into non-transfection group and transfection group, with 12 wells in each group. Cells in non-transfection group were not transfected with Ad-hEGF gene, while cells in transfection group were transfected with Ad-hEGF gene in the optimum MOI (50). The expression levels of cytokeratin 14 (CK14) and CK19 of cells were measured by immunofluorescence staining at TH 24. (5) Cells were collected, grouped, and treated as in (4), with 6 wells in each group. At post scratch hour (PSH) 0 (immediately after scratch), 12, 24, and 48, the migration distance of cells was observed and measured with inverted phase contrast microscope. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, and LSD test.

RESULTS(1) At TH 24 and 48, morphology of cells in each transfection group and non-transfection group were similar. Compared with that in non-transfection group, the cell debris increased significantly in 200 fold transfection group at TH 72. At TH 24, 48, and 72, the expression of green fluorescent protein was not seen in cells of non-transfection group, whereas it increased in cells of transfection group over transfection time. (2) The transfection rate of Ad-hEGF gene of cells in each transfection group increased gradually over transfection time. At TH 72, the transfection rates of Ad-hEGF gene of cells in 50-200 fold transfection groups were all above 90%, while the transfection rates of Ad-hEGF gene of cells in non-transfection group, 5, and 20 fold transfection groups were (0.51±0.20)%, (62.44±6.23)%, and (75.00±5.43)% respectively, which were obviously lower than the rate in 50 fold transfection group [(93.12±2.55)%, with P values below 0.01]. The mass concentration of EGF in culture supernatant of cells in each transfection group increased gradually over transfection time. At TH 72, the mass concentration of EGF in culture supernatant of cells in 50 fold transfection group was obviously higher than that in each of the other groups (with P values below 0.01). The proliferation activity of cells in each group at TH 24 and 48 was similar (with P values above 0.05). At TH 72, the proliferation activity of cells in 200 fold transfection group was obviously lower than that in other groups (with P values below 0.05). (3) On culture day 1, the biological activity of EGF secreted by cells in two groups was similar (P>0.05). On culture day 3 and 5, the biological activity of EGF secreted by cells in transfection group were obviously higher than that in non-transfection group (with P values below 0.01). (4) At TH 24, the expression levels of CK14 and CK19 of cells in transfection group were higher than those in non-transfection group. (5) The width of scratch in two groups was nearly the same at PSH 0. At PSH 12-48, the migration distance of cells in transfection group was obviously longer than that in non-transfection group (with P values below 0.01).

CONCLUSIONSThe suitable range of MOI of hECs transfected with Ad-hEGF gene is 50-150, and 50 is the optimum. hECs transfected with Ad-hEGF gene with MOI 50 can effectively express the EGF gene and keep its good abilities of proliferation, differentiation, and migration, as well.

Adenoviridae ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; EGF Family of Proteins ; genetics ; metabolism ; Epidermis ; cytology ; Genetic Vectors ; Humans ; Keratins ; metabolism ; Male ; Transfection

OBJECTIVETo study whether As(2)O(3) has an apoptotic effect on human solid tumor cells, and the possible cellular and molecular mechanisms of this treatment using human esophageal squamous carcinoma cells (EC8712) as a model.

METHODSDNA microarray, biochemical and cytological analyses were used.

RESULTSThe growth and survival of EC8712 cells were markedly inhibited by As(2)O(3) treatment at a concentration of 1, 2 and 4 micromol/L. EC8712 cells were obviously arrested at G2/M phase with As(2)O(3) treatment and apoptosis induced at micromolar As(2)O(3) concentrations, as shown by morphology, histogram related nuclear DNA contents, and DNA gel electrophoresis. As(2)O(3) activated caspase-3, which might be involved in the process of As(2)O(3), induced apoptosis in EC8712 cells.

CONCLUSIONSAs(2)O(3) changes the expression of many genes at transcription level. The regulation of expression of many genes might be involved in the process of As(2)O(3) inducing apoptosis. These results suggest that As(2)O(3) can be clinically useful for solid tumor treatment.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Carcinoma, Squamous Cell ; drug therapy ; genetics ; pathology ; Cell Adhesion ; drug effects ; Cell Division ; drug effects ; DNA Fragmentation ; drug effects ; Dose-Response Relationship, Drug ; Esophageal Neoplasms ; drug therapy ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Microscopy, Electron ; Oxides ; pharmacology ; Tumor Cells, Cultured ; drug effects ; ultrastructure

Objective:To investigate the epidermal growth factor receptor (EGFR) mutation rate, mutation characteristics and distribution characteristics of different mutation types in patients with non-small cell lung cancer (NSCLC) in Fuyang of Yunnan province, to provide the clinical individualized targeted therapy of NSCLC in this region.Methods:A total of 328 NSCLC patients whose native place were Fuyuan and who underwent EGFR test in Fuyuan County People's Hospital in Yunnan Province from January 2018 to August 2020 were selected, and their clinical data such as gender, age, ethnicity, pathological type and the results of EGFR test were collected for statistical analysis.Results:The EGFR mutation rate of NSCLC patients was 40.55% (133/328). The EGFR mutation rate of female patients was higher than that of males ( P < 0.01). The EGFR mutation rate showed a downward trend with age, the EGFR mutation rate of patients ≤ 60 years old was higher than that of patients > 60 years old ( P = 0.014). The EGFR mutation rate in ethnic minority was not statistically different from Han nationality ( P = 0.789). The EGFR mutation rate of patients without smoking history was higher than that of patients with smoking history ( P<0.01). Patients with adenocarcinoma had a higher EGFR mutation rate than squamous cell carcinoma ( P = 0.002). The EGFR mutation rate in patients with stage Ⅰ-Ⅱwere higher than that in patents with stage Ⅲ-Ⅳ ( P = 0.013). The EGFR mutation rate in tissue samples were higher than that in peripheral blood samples ( P = 0.009). In 328 patients the EGFR single-point mutation rate was 24.70% (81/328), and the compound mutation rate was 15.85% (52/328); the common mutation rate was 17.07% (56/328), and the rare mutation rate was 23.48% (77/328). The top 5 mutation types were L858R (10.06%), G719X+S768I (7.32%), 19-Del (7.01%), G719X+L861Q (6.40%), and G719X (4.21%). In 133 patients with EGFR mutation, the proportion of patients with rare mutation [57.89% (77/133)] was higher than the proportion of patients with common mutation [42.11% (56/133)]. Conclusion:The EGFR mutation rates of female, adenocarcinoma, non-smoking and young NSCLC patients in Fuyuan area are high, and the rare mutation rate is high.

Objective To investigate the safety and effectiveness of perventricular device closure (PDC) for congenital ventricular septal defects (VSD) by a meta-analysis. Methods PubMed, The Cochrane Library, EMbase and China Biology Medicine (CBM) databases were searched for studies on PDC of VSD up to October 2018 by computer. We included studies only with more than 80% patients diagnosed with perimembranous VSD. A meta-analysis was performed to obtain pooled estimates of success rate and incidences of complications with 95% confidence interval (CI). Subgroup analysis stratified by percentage of perimembranous VSD was performed. Risk difference (RD) was used in the comparison of perioperative data with follow-up data. Results Fifteen studies (2 randomized controlled trials, 3 retrospective cohorts, and other 10 retrospective single-arm studies) involving 4 164 patients (3 848 patients with perimembranous VSD) were included in this meta-analysis. The pooled success rate was 95.4%. Incidences of residual shunt, aortic insufficiency, tricuspid insufficiency, and third-degree atrioventricular block were 5%, 0.1%, 3%, and less than 0.001% respectively. Improvements of residual shunt and aortic insufficiency were confirmed in follow-up patients. Conclusion PDC of congenital perimembranous VSDs is safe and yields good results. Because some limitations can not be overcomed, multicenter randomized controlled trials are needed to confirm our results.

Hepatocellular carcinoma(HCC)is one of the most common malignancies with high morbidity and mortality rates worldwide.With the advances in molecular biology and tumor immunology,molecular-targeted agents represented by tyrosine kinase inhibitors(such as sorafenib and lenvatinib)and immunotherapy represented by PD-1/PD-L1 monoclonal antibodies have brought hope for patients with advanced HCC.The combination of immunotherapy and anti-angiogenic therapy can further improve the treatment outcome of patients.In addition,the optimization and integration of stereotactic body radiotherapy,local treatment,and systemic treatment may maximize the benefits of patients.In the future,through a deep understanding of the heterogeneity of HCC,the development of precision molecular subtyping and individualized treatment,and the establishment of a multidisciplinary collaborative diagnosis and treatment system,systemic therapy is expected to achieve long-term management of advanced HCC.This article reviews the current status and advances in systemic therapy for advanced HCC.