[Objective] To explore the effects of angongniuhuang pill on the synthesis of apolipoprotein E in brains of rats after traumatic brain injury.[Methods] Using reverse transcription-polymerase chain reaction(RT-PCR) and enzyme linked immunosorbent assay(ELISA),the apolipoprotein E’s mRNA of contused brain tissue and the concentration of apolipoprotein E in CSF at different phases after traumatic brain injury were detected of rats fed with angongniuhuang pills.[Results]High expression of apolipoprotein E mRNA and high concentration of apolipoprotein E were found in rats fed with angongniuhuang pills compared with controls.[Conclusion]Angongniuhuang pills may increase the expression of apolipoprotein E mRNA in brains after traumatic brain injury,thus promote the synthesizing and secreting of apolipoprotein E.

Objective:To analyze the influence of clinicopathological features on the prognosis of anorectal malignant melanoma (AMM), and to establish a more accurate prognosis prediction model.Methods:From January 1, 2006 to December 31, 2018, at Fudan University Shanghai Cancer Center, the data of 89 patients diagnosed with AMM and underwent operation were retrospectively analyzed. The Cox proportional hazard regression model was used to analyze factors affecting the prognosis. Backward elimination was used to select variables, Nomogram prognosis prediction model was established and verified internally, and the consistency index was calculated.Results:Among 89 AMM patients, 65 (73.0%) were female, 78(87.6%) were <70 years old, and the most common tumor location was the rectum (48.3%, n=43), followed by the anal canal (31.5%, n=28) and the anorectal canal (20.2%, n=18). Thirty-eight (42.7%) patients directly received abdominal-perineal resection (APR), 37 patients (41.6%) received interferon-based immunotherapy, and 11 patients (12.4%) received both systemic chemotherapy and immunotherapy. The 3-year disease-specific survival (DSS) rate was 41.6%(37/89), and the 5-year DSS rate was 31.5%(28/89). The results of multivariate Cox analysis showed that age ≥70 years old and T2 stage were risk factors of the prognosis of AMM patients (hazard ratio ( HR)=11.29, 4.83; 95% confidence interval ( CI) 2.89 to 44.13, 1.66 to 14.11; both P<0.01), while neurovascular invasion, immunotherapy treatment, systemic chemotherapy combined with immunotherapy, APR after extensive local resection, directly APR and APR combined with inguinal lymphadenectomy were protective factors of the prognosis ( HR=0.09, 0.23, 0.10, 0.13, 0.26, 0.02; 95% CI 0.02 to 0.34, 0.10 to 0.57, 0.02 to 0.49, 0.03 to 0.52, 0.08 to 0.90, 0.00 to 0.27; all P<0.05). The Nomogram model was further established with age, gender, tumor location, T stage, distant metastasis, medication chemotherapy and surgical treatment. The results of the Nomogram model internal verification indicated that the accuracy of the model in predicting 1-year, 3-year and 5-year DSS was good, and the consistency index was 0.749, which was significantly higher than the consistency index of traditional TNM stage (0.607). Conclusions:Most AMM patients are <70 years old, and the majority of them are female. The common location of AMM is rectum, and many patients receive immunotherapy. Age ≥70 years old and T2 stage are risk factors affecting the prognosis of AMM patients. Neurovascalar invasion, immunotherapy based comprehensive treatment, APR after extensive local resection, directly APR and APR combined with inguinal lymphadenectomy are protective factors of the prognosis. Nomogram prognosis prediction model established based on the clinicopathological features and treatment of AMM patients has higher accuracy and clinical reference value than the traditional TNM stage system.

Objective To acquire the physiological parameters such as pulse,blood oxygen saturation and ECG to realize personal health information monitoring and tailored medical service.Methods A personal health information acquisition terminal was developed,which was composed of an information acquisition module,personal computer and system software.The information acquisition module was developed with USB interface and unified circuit mode.Health information was transmitted to the personal computer at real time with USB interface,and then stored by system software and uploaded to the database.Results The terminal gained advantages in design,convenience and practicability,which realized realtime acquisition,data storage,remote auxiliary diagnosis,printing and etc of personal health information,so that personal information realtime monitoring was implemented.Concluslon The terminal can be used for health information recording,storage,acquisition and analysis,and extends medical service of hospital.

Objective To explore the mechanism of hBUB1 gene in the developing of spontaneous abortion embryos with numerical chromosomal abnormalitv.Methods Quantitative real-time RT-PCR and Western blot were used to determine the mRNA and protein level of hsMAD2 gene both in spontaneous abortion embryos with numerical chromosomal abnormality(experimental group)and with numerical chromosomal normality(control group).Recombinant shRNA plasmids targeting hBUB1 gene was constructed to inhibit the expression of endogenous hBUB1 genes in embryonic cells.Interference efficiency was demonstrated by fluorescent quantitative PCR and Western blot.The inhibitory rate of cell proliferation was measured by MTT assay and cells-cycle was assessed by flow cytometry.Resuns Western blot analysis showed that protein level of hBUB1 in the experimental group was decreased signifieandv(the rate of positivity and strong positivity were 8%and 93.5%,respectively,P＜0.05)compared with the control group.The expression of hBUB1 gene in embryonic cells was significantly and specially inhibited by shRNA plasmids (the mRNA level before and after treatment witll RNAi were 0.196±0.067 and 0.042±0.006,respectively,P＜0.05).The inhibitory rate of cell proliferation was increased to 62%from 4%at 48 h after transfection.The rate of G2/M phase cells was decreased after transfection with efficient shRNA(control group:40.2%and 41.3%,test group:21.3%).Conclusions Down-regulation of hBUB1 gene leads to the inhibition of cell proliferation and the arrest of cell-cycle.It also probably plays an important role in the development of spontaneous abortion embryos with numerical chromosomal abnormality.The clinical relevance warrant further investigation.

Objective To investigate the effect of fluvastatin (FLV) on the expression of β1 integrin in puromycin aminonucleoside (PAN)-treated podocytes and its mechanism.Methods Cultured human podocytes were divided into PAN,different concentrations of fluvastatin (1 × 10-8 to 1 × 10-5 mol/L),SOD,H2O21 groups respectively.Expressions of β1 integrin and reactive oxygen species (ROS) in podocytes were detected by Western blotting and DCFHDA (2' 7'-Dichlorofluoresecein 3' 6'-diacetate) respectively.The viability of podocyte was determined by MTT colorimetry.Results PAN and H2O2 significantly decreased the expression of β1 integrin and increased the synthesis of ROS in podocytes (P＜0.05respectively).Lower concentration fluvastatin or SOD treatment up-regulated β1 integrin and downregulated ROS of podocytes induced by PAN (P＜0.05 respectively).MTT revealed that lower podocyte viability was found in higher concentration fluvastatin,PAN and H2O2 groups.Lower concentration fluvastatin and SOD could protect podocytes against PAN.Conclusion Fluvastatin attenuates the injury of podocyte induced by PAN and increases the expression of β1 integrin,whose mechanism may be associated with the inhibition of the ROS activity.

Objective To explore the effect and mechanism of fluvastatin on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS).Methods Cultured HPMCs were randomly divided into control,HGPDS,HGPDS plus GSK650394 10-5 mol/L (the competitive inhibitor of SGK1),different concentrations of fluvastatin,fluvastatin 10-6 mol/L and GSK650394 10-5 mol/L alone.The morphology change of HPMC was observed by light microscopy.The cellular viability was detected by MTT colorimetry.The mRNA and protein expressions of serum and glucocorticoid-inducible kinase 1 (SGK1) and FN were detected by RT-PCR,Western blotting or ELISA.Results After incubation with HGPDS,the cell morphology changed from typical cobblestone-like appearance to fibroblast-like appearance,and the cell viability was inhibited significantly (P＜0.05).Fluvastatin 10-6mol/L and GSK650394 could improved the cell morphology and the cell viability injured by HGPDS (P＜0.05).Compared with the normal control group,the mRNA and protein expressions of SGK1 and FN increased significantly in HPMC treated with HGPDS(P＜0.05).GSK650394 significantly decreased the high expression of SGK1 and FN (P＜0.05),also the fluvastatin had same effects as GSK650394 in dose-dependent manner (P＜0.05).Conclusions High-glucose peritoneal dialysate can increase FN expression in human peritoneal mesothelial cells,which can be attenuated by fluvastatin.The protective role of fluvastatin in HPMC may be partially achieved through the signal pathway of SGK1.

Objective To study the diagnostic value of combined detection of serum CEA,CA199,CA125,AFP and fecal occult blood(FOB) in Dukes B stage colon carcinoma.Methods The above indicators in 56 patients with colon carcinoma (colon carcinoma group),35 patients with colonic polyp benign disease and 41 healthy individuals undergoing physical examination were measured.Then the sensitivity,and specificity of single index detection and their combined detection were analyzed.Results The levels and positive rates of CEA,CA199,CA125 and FOB in the colon cancer group were significantly higher than those in the colonic polyp group and healthy control group (P＜0.05),but the AFP level and positive rate had no statistical difference compared with the colonic polyp group.Serum CEA,CA125,CA199 and FOB were selected into the Logistic regression equation.The corresponding areas under the curve (AUC) were 0.745,0.886,0.792 and 0.864 respectively;the positive detection rates were 48.2 ％,35.7 ％,39.3 ％ and 78.6 ％ respectively.The combined detection could increased the specificity,sensitivity and AUC to 87.5 ％,92.1％ and 0.957,which were more superior to that in the single index detection.Conclusion The combined detection of serum CEA,CA199,CA 125 and FOB can provide the powerful basis for early diagnosing colonic cancer.

Objective To explore the effect and associated mechanism of fluvastatin combined with losartan on the expression of vascular endothelial growth factor (VEGF) in human podocytes induced by angiotensin (Ang)Ⅱ.Methods The differentiated human podocytes were cultured with various concentrations of Ang Ⅱ (10-9 to 10-5 mol/L) in vitro,followed by treatment of fluvastatin (10-7,10-6 and 10-5 mol/L),losartan (10-7,10-6 and 10-5 mol/L),extracellular signal-regulated kinase (ERK) inhibitor PD98059,and combination of fluvastatin and lolsartan.Expressions of VEGF and phosphorylation (p)ERK1/2 protein in podocytes were detected by Western blotting.RT-PCR was used to examine VEGF mRNA expression (P＜0.01).Results Ang Ⅱ up-regulated the expressions of VEGF and p-ERK1/2 in time-and dose-dependent manner.Above elevated expressions of VEGF and p-ERK1/2 induced by Ang Ⅱ could be down-regulated by fluvastatin,losartan and PD98059 respectively (P＜0.05).More obvious reduction of above expressions was found in combination of fluvastatin and losartan as compared to single agent (P ＜0.05).Conclusions Either fluvastatin or losartan can down-regulate the over-expression of VEGF and p-ERK1/2 induced by Ang Ⅱ in human podocytes,and their combination has a cooperative effect.The ERK signaling pathway may be one of the mechanisms of their renal protective effects.

Objective Detecting of spermatozoal total RNAs by laboratory on chip gel electrophoresis so that it could provide better total RNAs for the sequent experiments, and spur the development of spermatozoal molecular biology. Methods Sperms of healthy adults were collected and then total RNAs were extracted by RNeasy mini kit(QIAGEN), detection and quality control were performed by loboratory on chip gel electrophoresis system. Meantime, the control RNAs were extracted from lymphocytes. Results It was found that there were a plenty of genes expressed in healthy sperms. Electrophoretic graphs showed that the total RNAs of spermatozoal had 2 bands which went ahead a little comparing to the normal somatic cells. The former peak appeared keenness, and the latter was broad and showed like a reversed U. The ratio of them was largely more than 2, no extra peaks were found in electrophoretic graph. Conclusion A simple,intuitionistic method to detect and control the quality of the healthy adults' spermatozoal total RNAs had been successfully constructed by using laboratory on chip gel electrophorosis.

Objective:To investigate the clinical efficacy of cyclophosphamide combined with leflunomide in the treatment of lupus nephritis.Methods:The clinical data of 90 patients with lupus nephritis who received treatment in The Second People's Hospital of Liaocheng from January 2019 to June 2020 were retrospectively analyzed. These patients were divided into two groups according to different treatment methods. The single drug group ( n = 45) was treated with cyclophosphamide alone, and the combined drug group ( n = 45) was treated with cyclophosphamide combined with leflunomide. All patients were treated for 6 months. Total response rate, inflammatory factor level, immune function, renal function, and adverse reactions were compared between the two groups. Results:Total response rate in the combined drug group was 95.56% (43/45), which was significantly higher than 82.22% (37/45) in the single drug group ( χ2 = 4.05, P < 0.05). After treatment, interleukin-6, C-reactive protein, and rheumatoid factor in the combined drug group were (45.21 ± 5.07) ng/L, (3.13 ± 1.01) mg/L, (43.37 ± 18.20) IU/mL, respectively, which were significantly lower than (60.20 ± 6.13) ng/L, (6.23 ± 1.31) mg/L, (73.19 ± 19.17) IU/mL in the single drug group ( t = 12.64, 12.57, 7.56, all P < 0.001). Immunoglobulin A and immunoglobulin G levels in the combined drug group were significantly lower than those in the single drug group ( t = 13.05, 13.40, both P < 0.001), but immunoglobulin M level in the combined drug group was significantly higher than that in the single drug group ( t = 13.51, P < 0.001). Serum creatinine and 24-hour urine protein levels in the combined drug group were (78.23 ± 19.13) μmol/L and (1.15 ± 0.33) g/24 hours, respectively, which were significantly lower than (92.19 ± 20.19) μmol/L and (3.15 ± 0.81) g/24 hours in the single drug group ( t = 3.36, 15.33, both P < 0.001). The incidence of adverse reactions in the combined drug group was 6.67% (3/45), which was significantly lower than 22.22% (10/45) in the single drug group ( χ2 = 4.40, P < 0.05). Conclusion:Cyclophosphamide combined with leflunomide is effective against lupus nephritis. The combined therapy can regulate the inflammatory reaction, improve the immune function, promote the recovery of renal function, and be safe.