AIM:To put forward three-level design(Box-Behnken design)for fitting response surfaces to optimize the extraction technology of total flavonoids from Hypericum ascyron L.METHODS:Four factors,including temperature,reflux time,concentration of ethanol,ratio of solvent to raw material were used to examine the yield of total flavonoids.Prediction was carried out through comparing the observed and predicted values.RESULTS:The results suggested that ethanol concentration and extraction temperature were two statistically significant factors.The optimum conditions of extraction process consisted of the ratio(mL:g)of solvent to material(13.3:1),ethanol concentration(53.2%),extraction temperature(78.7℃)and extraction time(2.3h).Regression coefficient of binomial fitting complex model was as high as 0.984 6.Bias between observed and predicted values was-4.01%.CONCLUSION:Box-Behnken design is success in optimizing the extaction in close agreement with the predicted values of the mathematic model.

Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma. Methods: Balb/c mice were immunized i. p . with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes ( VH and VL) of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA. , the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13K07 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3. 5 ? 106 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified , which paves a way for study of prostate carcinoma.

Objective:To obtain phage-displayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display,so as to lay a foundation for diagnosis and treatment of cancer.Methods:The membrane proteins were extracted from the specimens of esophageal carcinoma,stomach carcinoma,brain cancer,lung cancer,and spinal cord tumor.The recombinant phage-antibody system was used to construct a single-chain Fv fragment(ScFv)cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein.The specific primers of VH and VL were used to amplify the cDNA of VH and VL,respectively,which were then assembled into ScFv gene with a specially constructed linker DNA.The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1.The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma(HeLa cells),sepecific phage-ScFvs were selected by phage displaying and panning in vivo.After four rounds,24 phage-ScFvs,which were identified by PCR,were analyzed immunohistochemically.The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced.Results:Tomors-bearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice.It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice.A ScFv library of 1.6?106 was obtained and a tumor vessel specific phage-ScFv named ScFvH1(VH-linker-VL)was selected from the library.Conclusion:A tumor targeting ScFv library has been successfully constructed and a tumor vessel-specifrc antibody has been identified from the library,which provides a new way for the early diagnosis and therapy of cancer.

AIM:To put forward three-level design(Box-Behnken design) for fitting response surfaces to optimize the extraction technology of total flavonoids from Hypericum ascyron L.METHODS:Four factors,including temperature,reflux time,concentration of ethanol,ratio of solvent to raw material were used to examine the yield of total flavonoids.Prediction was carried out through comparing the observed and predicted values.RESULTS:The results suggested that ethanol concentration and extraction temperature were two statistically significant factors.The optimum conditions of extraction process consisted of the ratio(mL:g)of solvent to material(13.3:1),ethanol concentration(53.2%),extraction temperature(78.7℃)and extraction time(2.3h).Regression coefficient of bino-mial fitting complex model was as high as 0.984 6.Bias between observed and predicted values was -4.01%.CONCLUSION:Box-Behnken design is success in optimizing the extaction in close agreement with the predicted values of the mathematic model.

ObjectiveTo systematically review the clinical of artificial liver support system (ALSS) in the treatment of drug-induced liver failure. MethodsPubMed, Embase, The Cochrane Library, CMB, CNKI, and VIP databases were searched for related randomized controlled trials or randomized controlled trials on ALSS in the treatment of drug-induced liver failure published up to October 2019, and a statistical analysis was performed. Odds ratio (OR) was the effect size for categorical data, and the difference between groups was the effect size for continuous data. The weighted mean difference (WMD) method was used for the pooled analysis of effect size, and 95% confidence interval (CI) was calculated for each effect size. I2 and P values were used to evaluate the heterogeneity of the articles included in the analysis; a fixed effect model was used when I2＜50% and P＞0.1, otherwise a random effects model was used. ResultsA total of 16 articles with 945 patients were included, with 520 patients in the ALSS+routine medical treatment (RMT) group and 425 in the RMT group. The meta-analysis showed that compared with the RMT group, the ALSS+RMT group had a significantly lower mortality rate of drug-induced liver failure (OR=0.27, 95%CI: 0.20-0.36, P＜0.001), significant improvements in albumin (Alb) (MD=1.21, 95%CI: 0.18-2.25, P=0.02) and prothrombin activity (PTA) (MD=11.84, 95%CI: 6.34-17.35, P＜0.001), and a significant reduction in total bilirubin (TBil) (MD=-104.97, 95%CI: -163.63 to -46.30, P＜0.001). Further analysis of Alb, TBil, and PTA after the withdrawal of ALSS showed that ALSS significantly improved Alb (MD=1.74, 95%CI: 1.20-2.27, P＜0.001) and PTA (MD=4.45, 95%CI: 2.80-6.10, P＜0.001) and significantly reduced TBil (MD=-128.41, 95%CI: -217.22 to -39.59, P=0.005). ConclusionCompared with RMT alone, RMT combined with ALSS can significantly improve the main biochemical indicators of patients with drug-induced liver failure and reduce their mortality rate.

OBJECTIVE:To optimize the formulation of nanostructured lipid carriers loaded with lornoxicam (LN-NLC). METHODS:Emulsification-solvent evaporation method was used to prepare the LN-NLC. Using drug-lipid ratio,dosage of soy lec-ithin,liquid-lipid ratio (proportion of liquid lipid to total lipid) and dosage of emulsifier as factors,the overall normalized value was calculated by particle size,Zeta potential and entrapment efficiency as indexes was used as comprehensive index. Central com-posite design-response surface method was used to optimize the formulation and investigate the appearance and stability of prepared LN-NLC. RESULTS:The optimal formulation were as follows as drug-lipid ratio of 1:50,dosage of soy lecithin of 162.5 mg,liq-uid-lipid ratio of 25% and emulsifier dosage of 958.2 mg. The particle size of prepared LN-NLC was(96.9±3.3)nm,Zeta poten-tial was(-16.1±0.3)mV,entrapment efficiency was(60.1±0.9)%(n=3),which showed relative error of 2.47%,-4.55%,-0.17%with predicted value,respectively. The prepared LN-NLC was spherical. It had no obvious changes in particle size and Ze-ta potential in sealed storage for 30 d in 4 ℃,and the entrapment efficiency only declined 1.2%. CONCLUSIONS:The LN-NLC formulation is successfully optimized,and the LN-NLC has good stability.

Portal hypertension (PH) is a common clinical syndrome, and the main risk factor for death is esophageal variceal bleeding. Therefore, the early and accurate diagnosis of PH can reduce the incidence rate of serious complications and mortality rate. Hepatic venous pressure gradient, as a main method for the diagnosis of PH, has certain limitations in clinical application, and thus it is necessary to explore new noninvasive diagnostic techniques. Up to now, ultrasound elastography has become one of the important methods for the noninvasive assessment of PH. This article elaborates on its current application and prospects in the assessment of PH.

Objective To observe the effect of Rhy-SLN on the proliferation of rat vascular smooth muscle cells (VSMC) induced by TGF-β1, and explore the mechanism. Methods The primary culture of rat thoracic aortic vascular smooth muscle cells was studied by tissue block culture method. The cells were divided into the control group, TGF-β1 group, TGF-β1+ the high, medium and low dosage groups of Rhy-SLN. In addition to the control group, the cells of the other groups were involved in the intervention of TGF-β1 of 20 g/L, and the high, medium and low dosage groups of Rhy-SLN cells were involved in the intervention of 25, 50, 100 mg/L of the hook teng solid lipid nanoparticles. After 24 hours of culture, MTT assay was used to detect cell proliferation inhibition rate in each group, and the cell cycle was detected by flow cytometry. The expression of c-myc and c-Fos protein in each group was detected by Western blot method. Results Compared with the TGF-β1 group, the absorbance value (0.457 ± 0.046 vs. 0.975 ± 0.049) of TGF-β1+ rhy-sln high dose group significantly decreased (P<0.01); the number of S phase cells (15.87% ± 2.47%, 15.23% ± 1.69%, 17.02% ± 2.87% vs.38.58% ± 2.68%)of TGF-β1+rhy-sln in each dose group significantly decreased(P<0.01);The c-myc(48.65 ±3.65,50.69 ± 4.16,55.29 ± 3.67 vs.68.21 ± 3.25)and c-Fos(38.78 ± 4.25,43.56 ± 3.69,46.58 ± 3.57 vs.66.54 ± 4.09) of the TGF-β1+ rhy-sln each dose group significantly decreased (P<0.01). Conclusions The Rhy-SLN can inhibit the proliferation of VSMC in rats induced by TGF-β1.Its mechanism is related to the conversion of G0/G1 phase to the S phase and the expression of the reduction of c-myc and c-fos protein.

BACKGROUND: Mild cognitive impairment (MCI) in elderly people is becoming increasingly a new hot spot for the clinical study of Alzheimer disease at present, and it is still unknown how many cases of MCI will convert into Alzheimer disease. OBJECTIVE: To analyze the incidence rate of mild cognitive impairment and the conversion rates into dementia and Alzheimer disease among elderly people.DESIGN: A prospective cohort study based on diagnosis. SETTING: Second Department of Cadre's Ward, Bethune International Peace Hospital of Chinese PLA.PARTICIPANTS: The subjects were 216 MCI patients and 2 302 subjects with normal cognition selected in the baseline investigation to the retired cadres aged 60 years and above, in 26 military cadres' sanatoriums of Shijiazhuang city between August and September 2001.METHODS: The MCI diagnosis was according to the MCI diagnostic standard in Diagnostic and Statistical Manual of Mental Disorders - Fourth Edition (DSM-Ⅳ), and the diagnosis of dementia was according to the standard in DSM-Ⅳ revised by American Psychiatric Association. Alzheimer disease was diagnosed according to the standards of National Institute of Neurological and Communicative Disorders and Stroke/the Alzheimer Disease and Related Disorders Association (NINCDS-ADRDA). All the 216 MCIpatients and 2 302 subjects with normal cognition were enrolled in the 3-year cohort study. MAIN OUTCOME MEASURES: The incidences of MCI among subjects with normal cognition; the annual conversion rates into Alzheimer disease in MCI patients and subjects with normal cognition; the relative risks (RR) and 95% confidence interval (95% CI) of MCI and normal cognition developing into Alzheimer disease. RESULTS: The MCI group and normal cognition group were followed up once every year for 3 years. Of the 216 MCI patients, 7 died, and 209 were followed up actually, and the follow-up rate was 96.8%. Of the 2 302 subjects with normal cognition, 36 died, and 2 266 were followed up actually,and the follow-up rate was 98.4%. ① In the elderly subjects with normal cognition, the annual incidence of MCI was 4.8%, and those of dementia and Alzheimer disease were 1.3% and 0.8 respectively. ② The annual in cidence rates of dementia and Alzheimer disease in MCI patients were 8.1% and 5.6% respectively, which were insignificantly different between males and females (P ＞ 0.05). ③ The incidence rates of dementia and Alzheimer disease showed a descending trend with the elevation of educational level in both the MCI patients (trend x2 =5.57, P=0.02; trend x2 =4.92, P=0.03) and the subjects with normal cognition (trend x2 =23.1, P＜ 0.001; trend x2 =18.0, P ＜ 0.001). ④ The annual incidence rates of dementia and Alzheimer disease showed an ascending trend with aging in both the MCI patients (trend x2 =14.6, P ＜ 0.01; trend x2 =13.9, P ＜ 0.01) and the subjects with normal cognition (trend x2 =32.3, P ＜ 0.01; trend x2=23.8, P ＜ 0.01). ⑤ The relative risks of the conversion into dementia and Alz heimer disease in MCI patients were 6.4 and 7.4 times as many as those in the subjects with normal cognition.CONCLUSION: The risks of conversion into dementia and Alzheimer disease in elderly MCI patients are far higher than those in elderly people with normal cognition. It should be reinforced to monitor elderly MCI patients, who are the of high risk group of dementia.

Objective To explore the feasibility of the extended inferior meatal maxillary antrostomy through inferior turbinate reversing approach.Methods Eight patients underwent extended inferior meatal maxillary antrostomy through inferior turbinate reversing approach for maxillary sinus lesions.The indications,surgical management and outcomes of the surgery were presented.Results There were 2 cases of maxillary dentigerous cyst with oroantral fistula,2 cases of antrochoanal polyp,2 cases of maxillary sinus inverted papillomas,1 case of odontogenic maxillary sinusitis with oroantral fistula,and 1 case of maxillary sinus mucocele.All patients underwent extended inferior meatal maxillary antrostomy through inferior turbinate reversing approach without intraoperative complication,with good access to the lesions.Complete resection could be achieved through this approach,no postoperative complications occurred except one patient had a delayed wound healing of inferior turbinate,all patients were free of recurrence with the average postoperative follow-up of 7.8 months (range 4-12 months).Conclusion The extended inferior meatal maxillary antrostomy through inferior turbinate reversing approach is recommended for some maxillary pathology owing to its good access to the lesions and complete resection.