Objective:Antimicrobial peptides ( AMPs ) are peptides generated in the biological defense system against exogenous pathogens ,which is all important constituent of the non-specific immune system.The immune system can react and sometimes play an important role in tumor control both in animal models and in humans.In this study,CecropinXJ were used in the BALB/c Eca109 cells-bearing mice model.To investigate whether CecropinXJ exert anti-tumor efficiency through regulating immune response when treated with tumor-bearing mice.Methods:The expression of cytokines and DC surface molecules were detected by immunohisto-chemistry and Flow cytometry.Results:The experiment of mice showed that CecropinXJ could significantly inhibit the proliferation of transplanted tumors in dose-dependent manner.There were significant difference between control group and 10mg/kg CecropinXJ group on the volume and weight of tumor ( P<0.05 ).CecropinXJ inhibited Eca 109 cells infiltrating growth in tumor tissue without affecting viscera,and the expressions of VEGF and bFGF proteins were higher than those in the control group .The expression of cytokines and DC surface molecules,the results that,compared with the control group,CecropinXJ could not significantly promote the expression of CD4,IL-4 and IFN-γ,while not involving the expression of surface molecules on DCs in spleen (P>0.05).In CecropinXJ treatment group,TNF-αlevel in serum was significantly higher than the control group ( P<0.01 ).Conclusion: CecropinXJ could significantly inhibit tumor growth in Eca109 cells-bearing mice,and might not be affected anti-tumor efficiency through regulating immune response.

Objective:To observe the clinical efficacy of Dushen Decoction and Zhenwu Decoction in the treatment of chronic congestive heart failure (CCHF) of Yang deficiency and blood stasis type. Methods:From March 2017 to December 2019, 120 patients with CCHF of Yang-deficiency and blood stasis type admitted to Shanghai Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine were selected, and they were divided into study group and control group according to the randomized number table method, with 60 in each group. The control group was given western medicine of conventional treatment, and the study group was combined with Dushen Decoction and Zhenwu Decoction on the basis of the control group. Both groups were treated for 1 month. Before and after treatment, TCM syndrome scores were performed, serum TNF-α, IL-17 and CRP were detected by ELISA, left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVEDD) and left ventricular end systolic diameter (LVESD) were detected by color doppler echocardiography the adverse reactions during the treatment were recorded and the clinical efficacy was evaluated. Results:The total effective rate was 91.7% (55/60) in the study group and 71.7% (43/60) in the control group, with significant difference between the two groups ( χ2=8.015, P=0.005). After treatment, the scores of palpitation, dyspnea, chilly limbs, dull tongue and edema in the study group were significantly lower than those in the control group ( t=13.953, 13.915, 30.945, 32.339, 20.403, P<0.001). After treatment, LVEF [(56.28 ± 4.34)% vs. (42.47 ± 4.56)%, t=16.993] in the study group was significantly higher than that of the control group ( P<0.01); LVEDD [(44.32 ± 6.23) mm vs. (53.81 ± 5.19) mm, t=9.066] and LVESD [(31.28 ± 4.62) mm vs. (37.51 ± 4.73) mm, t=7.299] in the study group were significantly lower than those in the control group ( P<0.01). After treatment, the serum levels of TNF-α, IL-17 and CRP in the study group were significantly lower than those in the control group ( t=12.644, 15.975 and 14.379, P<0.001). The incidence of adverse reactions was 6.7% (4/60) in the control group and 8.3% (5/60) in the study group, and there was no significant difference between the two groups ( χ2=0.120, P=0.729). Conclusion:Dushen Decoction and Zhenwu Decoction in the treatment of CCHF can improve the clinical symptoms, improve the cardiac function, reduce inflammatory factors, improve the treatment efficiency with good safety.

Objective:Primary gastric choriocarcinoma (PGC) is a rare tumor. This study aimed to analyze cases reported in Chi-nese and two cases in our hospital. This study was also conducted to investigate the clinical characteristics and treatment of PGC. Meth-ods:The Chinese Medical Current Contents and China Biology Medicine database was retrieved with the following terms:primary gas-tric choriocarcinoma, stomach choriocarcinoma, and ectopic choriocarcinoma. The clinical data in published literature were retrospec-tively analyzed. Two cases from our hospital were also retrospectively analyzed. Results:A total of 16 patients with PGC (14 case stud-ies and 2 cases from our hospital) were included in this study. The average age of the patients was 57.69 years (ranging from 42 years to 79 years). Among the 16 patients, 4 were females and 12 were males. The main clinical manifestation was abdominal discomfort (12/16). Laboratory and imaging tests were non-specific, but HCG-βin all of the patients increased (11/11). Organ or lymph node metasta-sis was found in 13 patients, including 6 cases in which two or more organs showed metastasis. These organs were primarily the liver, the stomach lymph nodes, the lungs, the brain, and the abdominal cavity. Pathological examination results showed that 12 of the 16 cas-es displayed ulcers with necrosis and hemorrhage. The four remaining cases manifested lesions. Lesions may occur in the stomach less-er curvature (5 cases), stomach sinus (6 cases), cardiac (2 cases), and gastric body (3 cases). The histological components of PGC co-ex-isted with other factors, such as adenocarcinoma (7 cases), adenocarcinoma and squamous (2 cases), adenocarcinoma and a signet ring cell (1 case), or pure choriocarcinoma (6 cases). Surgery combined with chemotherapy was the main treatment. A total of 14 patients un-derwent surgical operations. Among these patients seven underwent chemotherapy after surgery and one underwent chemotherapy. Among the 10 patients who were followed up, 8 died within six months after diagnosis. Conclusion:PGC is a rare stomach cancer with poor prognosis and primarily affects older men. No characteristic clinical manifestations were observed, but HCG-βas a specific serum tumor biomarker increased in all of the affected patients. Histological components co-existed with other stomach cancers. Surgical exci-sion combined with chemotherapy was the preferred treatment in early stages, and systemic chemotherapy was the recommended treat-ment in advanced stages.

Objective To compare Montreal cognitive assessment-basic ( MoCA-B ) and mini-mental state examination (MMSE) in screening cognitive dysfunction of acute stroke patients. Methods The cognitive function of patients (n=83) with acute stroke onset within 10 days (including new cerebral infarction and cerebral hemorrhage) were assessed using MMSE and MoCA-B. The classification of patients with cognitive impairment was compared between the two scales. The consistency of cognitive impairment and affected domains assessed by MMSE or MoCA with experts were evaluated. Results ①There were 32 cases (38.6%) with abnormal MMSE score and 40 cases (51.8%) with abnormal MoCA-B score. ②The the diagnostic consistency of MoCA-B with experts was 89.16%. The false positive of MMSE was 2.41%and the false negative (rate of missed diagnosis) was 16.87%.False positives of MoCA-B were 4.82%and false negatives (rate of missed diagnosis) were 6.02%.③Among the 51 patients with normal MMSE, 15 had abnormal MoCA-B (29.4%). There were significant differences between these two score system in executive function, verbal fluency, directivity, abstraction, delayed recall, visual perception, naming and other cognitive domains (P＜0.05). Conclusion MoCA-B scale may be more sensitive and better than MMSE scale in screening for cognitive impairment in patients with acute stroke.

Objective To assess the association of synovial cyclic citrullinated peptide(CCP)expression with T helper 17(Th17) cells/Regulatory T cells (Treg) imbalance,the histological and clinical features of synovitis in rheumatoid arthritis (RA).Methods CCP expression in synovial specimens from 39 patients with RA and 35 controls was detected by immunohistochemistry assay(IH) using 6×His tagged anti-CCP single chain fragment V (ScFv) antibodies,which were generated by pHEN2 phagemid recombinant antibodies display system.The frequencies of Th17/Treg cells in the peripheral blood mononuclear cells (PBMCs) were determined by flow cytometry (FCM).Th17/Treg cells associated cytokines were analyzed by enzyme-linked immunosorbent assay (ELISA).The histological scores and clinical features of synovitis were included in the study.Chi-square test and ANOVA were used for statistical analysis.Results ① The prevalences of synovial C CP expression were significantly different between RA group and the control(76.9％ and 11.4％ respectively,X2=31.9,P＜0.01).② The frequencies of Th17 cells,Th17/Treg ratio,Th17 cells associated cytokines as IL-6,IL-17a,IL-23,TNF-α,and the Treg cells associated cytokines TGF-31,the serum and synovial fluid anti-CCP antibodies in the RA patients with synovial CCP positive expression were significantly higher in RA patients with CCP positive than those with CCP negative.Disease activity score DAS28 index and histological features quantified variations of the synovial biopsy specimens (synoviocyte hyperplasia,focal aggregates of lymphocytes,and diffuse infiltrat(e)s of lymphocytes) in RA were higher in synovial CCP positive expression patients than in the negative.Conclusion Synovial CCP expression is strongly associated with the Th17/Treg imbalance and synovitis,which may play a crucial role in the pathogenesis of RA.

Objective:To investigate the short-term clinical efficacy of laparo-gastroscopic esophagectomy (LGE).Methods:The retrospective and descriptive study was conducted. The clini-copathological data of 11 patients with esophageal cancer who underwent LGE in the Zhongshan Hospital of Fudan University from June 2020 to October 2021 were collected. There were 8 males and 3 females, aged (68±4)years. Sorted by operation time, the sentinel lymph nodes navigation (SLN) was performed since the sixth patient in the cohort, and abdominal surgery and neck surgery were performed simultaneously to complete LGE. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect death of patients during postoperative 30 days. Patients were followed up during postoperative 30 days. Measurement data with normal distribution were represented as Mean± SD, and count data were described as absolute numbers. Results:(1) Surgical situations. Of the 11 patients, 5 cases received SLN with satisfactory visualization, 6 cases did not receive SLN, 1 case terminated the operation as sentinel lymph nodes biopsy showing positive results and the rest of 10 cases completed LGE successfully without conversion to thoracotomy. The operation time and tumor diameter of the 10 patients completing LGE was (204±27)minutes and (2.5±1.0)cm, respec-tively. (2) Postoperative situations. Of the 10 patients completing LGE, 2 cases had pulmonary complications after surgery and recovered well with symptomatic treatment, and none of patient had anastomotic leakage or other serious complication. Results of postoperative histopathological examination showed squamous cell carcinoma in the 10 patients completing LGE. Nine patients were classified as T1b?3N0M0 stage and 1 patient was classified as T1bN1M0 stage. Ten patients completing LGE had R 0 resection and the number of lymph nodes dissected was 14±4. There were 3 cases with nerve bundle invasion, 2 cases with vascular invasion and 5 cases without nerve bundle and vascular invasion. The postoperative treatment time at intensive care unit and duration of hospital stay of the 10 patients completing LGE were (4.0±2.4)days and (7.2±1.5)days. (3) Follow-up. The 10 patients completing LGE were followed up and none of them died during the postoperative 30 days. Conclusions:LGE is safe and feasible. Combined with SLN can guarantee the oncology effect of surgery.

Objective To investigate the regulating effects of Krüppel-like factor 6 (KLF6) on the apoptosis of human lens epithelial cells (HLECs) by activating transcription factor 4 (ATF4) pathway and explore the bio-molecular mechanism of KLF6/ATF4-induced HLECs apoptosis.Methods HLECs (HLE-B3) were cultured using high glucose DMEM medium.The eukaryotic expression plasmid pEGFP-C2-ATF4 was transfected into the cells by liposome 2000 in the ATF4-transfected group,and pEGFP-C2 was transfected in the empty plasmid group.Then the cells were exposed to 20 mJ/cm2 ultraviolet ray B (UVB) for 200 seconds,The morphological changes of the cells were observed by hematoxylin & eosin staining and Hoechst33258 fluorescein staining.Cultured cells were transfected using pEGFP-C2-KLF6 and pEGFP-C2 plasmid and pSilencer-KLF6 (siKLF6) and pSilencer plasmid,respectively,and the expression of ATF4 protein in the cells was detected by Western blot assay.Culture cells were divided into four groups.pEGFP-C2 and pSilencer plasmids were co-transfected into the cells in the empty plasmid group;pEGFP-C2-KLF6 and pSilencer empty plasmid were co-transfected into the cells of the KLF6 + pSilencer group;pEGFP-C2 empty plasmid and pSilencer-ATF4 were co-transfected in the cells of the siATF4 + pEGFP-C2 group;pEGFP-C2-KLF6 and pSilencer-ATF4 plasmids were co-transfected in the cells of the KLF6 + siATF4 group,and then the cells were exposed to UVB.The apoptosis of the cells were detected by ELISA assay.Results Cultured cells grew well in the normal control group with the uniform morphology and regular arrangement.The karyopyknosis,karyorrhexis and enlargement of intercellular space were found in the cells exposed to UVB.In the ATF4 transfected group,the number of cells was decreased.The relative expression level of the ATF4 protein in the cells was 0.99±0.06 and 0.13±0.02 in the UVB+ATF4 transfected group and UVB+pEGFP-C2 plasmid group,respectively,with a significant difference between them (t =23.13,P＜0.01).The relative expression levels of KLF6 and ATF4 proteins in the KLF6 transfected group were higher than those in the empty plasmid group,and the relative expression levels of KLF6 and ATF4 proteins in the siKLF6 group were significantly lower than those in the empty plasmid group (all at P＜0.01).ELISA assay showed that the apoptotic rate in the ATF4 transfected group was 1.37± 0.11,which was significantly higher than 0.31 ±0.11 in the normal control group (t =8.034,P =0.001);the apoptotic rate of the cells was increased in the KLF6+pSilencer group and decreased in the siATF4+pEGFP-C2 group in comparison with the empty plasmid group (P＜0.01,P=0.02).In addition,the apoptotic rate in the KLF6+ siATF4 group was remarkably lower than that in the KLF6 + pSilencer group (P＜ 0.01).Conclusions KLF6 promotes the apoptosis of HLECs induced by UVB radiation.Silence of ATF4 gene reduces the apoptotic rate of the cells.ATF4 is probably a target factor in the regulating oathwav of KLF6 to apoptosis.

Objective To explore the role of forkhead box F2 (FoxF2) in the extracellular matrix of trabecular meshwork.Methods The cultured human trabecular meshwork cells (HTMCs) were divided into Scramble control group and FoxF2 small hairpin RNA (shRNA) group,then FoxF2 shRNA,the FoxF2 restructuring interference carrier was built,HTMCs were infected with FoxF2 shRNA lentivirus.Western blot assay was used to detect the expression of FoxF2 protein and extracellular matrix.Furthermore,Transwell counting experiment was used to analyze the migration ability of HTMCs.Results The cultured HTMCs grew well and showed a long spindle shape.The growth status of HTMCs was well,and their morphological characteristics were consistent with the HTMCs in vivo.The relative expression level of FoxF2 protein in the FoxF2 shRNA group was lower than that in the Scramble control group,with a significant difference between them (0.72 ± 0.02 vs.1.27 ± 0.05;t =16.68,P ＜ 0.01).The relative expression level of fibronectin (FN),collagen type Ⅰ (COL Ⅰ) and α-smooth muscle actin (α-SMA) were 0.43±0.03,0.53 ±0.08 and O.86±0.15 in the FoxF2 shRNA group,and 0.87±0.04,1.66±0.06 and 1.73 ±0.13 in the Scramble control group,respectively,the relative expression levels of FN,COL Ⅰ and α-SMA in the FoxF2 shRNA group were significantly lower than those in the Scramble control group (t =15.08,18.81,7.50,all at P＜0.01).The migration number of HTMCs in the FoxF2 shRNA group was significantly lower than that in the Scramble control group (117.30±11.41 vs.251.00±10.37;t =8.72,P＜0.01).Conclusions The FoxF2 shRNA lentivirus are successfully constructed,which can decrease the expression of FoxF2 in HTMCs.Low expression of FoxF2 can reduce the expression level of extracellular matrix protein in HTMCs and inhibit the migration ability of HTMCs.

Objective To observe the inhibitory effect of lentiviral vector miR-191 (LV-191) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR).Methods Eighty healthy 7-day-old C57BL/6J mice were randomly divided into 5 groups including normal group, non-intervention group, normal saline (NS) group, LV-191 group and LV-green fluorescent protein (GFP) group, 16 mice in each group. The OIR model was established in the non-intervention group, NS group, LV-191 group and LV-GFP group. NS group, LV-191 group and LV-GFP group were given an intravitreal injection of 1 μl of NS, LV-191 and LV-GFP at the age of 12 days. No injection was performed in the non-intervention group. In normal group,newborn mouse were maintained in room air form P0 to P17, and no treatment was performed. Mice in all five groups were euthanized at P17. Retinal neovasculation (RNV) was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR (RT-PCR) to detect miR -191 and P21 expression of retinal tissue.Results In the LV-191 group, the non-perfusion area were both significantly smaller than those in non-intervention group, NS group and LV-GFP group (F=127.20,P<0.001). The number of pre-retinal neovascular cell nuclei in retinas from LV-191 group were obviously lower than those in the retinas from non-intervention group, NS group and LV-GFP group (F=31.71,P<0.05). RT-PCR showed that the LV-191 and P21 level of LV-191 group increased significantly than other groups (F=10.95, 15.60;P<0.05).Conclusion Intravitreal injection of LV-191 inhibits RNV in mice model of OIR possibly through up-regulating p21.

Objective To observe the effect ofpolypyramidine tract binding protein-associated splicing factor (PSF) towards advanced glycation end products (AGEs) induced the apoptosis of Müller cells in vitro.Methods Experimental study.Müller cells were cultured and divided into groups according to the project design,plasmid enhanced green fluorescent protein-PSF were transfected into the cells to achieve the overexpression of PSF Müller cells in vitro,then cells were exposed to AGEs and the Morphological changes were observed by HE staining and Hoechst 33258 staining while the survival rate of cells were detected by MTT assay.The effects of PSF on AGEs-induced Müller apoptosis was measured by Cell Death Detection ELISA kit.Meanwhile,2',7'-diehlorofluorescin diaeetate staining was performed to monitor the protective effects of PSF on AGEs-induced Müller cells ROS.Results The morphology of cells in normal group was full and the cytoplasm staining was uniform.In N+AGEs group and Vec+AGEs group,cell volume decreased,cytoplasm was dense and concentrated,and eosinophilic staining was enhanced.The cell morphology of PSF+AGEs group was still full,with uniform cytoplasm staining and uniform nucleus staining.The viability of N+AGEs group,Vec+AGEs group and PSF+AGEs group were 0.42±0.11,0.35±0.12 and 0.68±0.12.The apoptosis values were 1.08 ± 0.16,0.96± 0.20 and 0.44± 0.08.The intracellular ROS levels were 28 833.67± 3 550.06,28 356.67±4 854.81,186 163.00±382.54.Compared with N+AGEs group and Vec+AGEs group,the cell viability of PSF+AGEs group was significantly improved (F=20.65,P=0.000),ce11 apoptosis value (F=43.43,P=0.000) and intracellular ROS level (F=1 8.86,P=0.000).Conclusion PSF overexpression play a protective role in AGEs-induced apoptosis by inhibiting the production of ROS in Müiller cells.