Objective To discuss the feasibility of computerized tomography (CT) calibration technique for the quantification of the hepatic fat content in vivo . Methods Eleven healthy volunteers, 53 patients with fatty liver, 10 cases of chronic hepatitis B and 10 cases of liver cirrhosis were tested with CT calibration test tubes. Test tubes contained solutions with Terbutyl alcohol, which CT scan density (CTD) is close to fat tissue, and 18.5% glucose solution which CTD is almost identical to normal liver. Variable Terbutyl concentrations of 10%, 20%, 30% and 40%, which stands for various fat content in liver, were used to measure patients with variable degrees of steatosis under CT scan. Results There was a negative linear correlation ( r = -0.982 , P

Objective　To study　how　ConA actives macrophages in vivo to produce cytotoxic effectors and its phagocytic functions,and　 cytotoxicity. Methods　 ConA was intraperitoneally injected(ip). Cock red blood cells(cRBC) were used to evaluate MΦ phagocytic activity,and S180 cells as target cells to analyze MΦ dependent cytotoxicity(MTC).Nitric oxide(NO),TNF-α　and IL-1 levels of MΦ cultural supernatant were measured using griess reagent,L929 cells MTT method and thymocytes proliferation test respectively. Results　 ConA could promote MΦ　to phagocytize cRBC and kill S180 cells,enhance the production of such factors as NO,TNF-α　and IL-1 by MΦ. There was significant difference compared with PBS　control group(P＜0.01). Conclusions　 ConA could stimulate MΦ to produce effectors, which mediate immune regulation of ConA to MΦ.

Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.

Objective:To investigate the clinical efficacy of bilateral route minimal- incision necrosectomy combined with continuous lavage for the treatment of infected necrotizing pancreatitis (INP).Methods:The retrospective and descriptive study was conducted. The clinical data of 20 patients with IPN who were admitted to Daping Hospital, Army Medical University from April 2016 to July 2019 were collected. There were 11 males and 9 females, aged (42±9)years. All the 20 patients underwent bilateral route minimal-incision necrosectomy, and then be continuous perfused and drainage within the purulent cavity postoperatively. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up using outpatient examination and telephone interview was performed to detected patients fever, abdominal pain, abdominal distension, diarrhea, peripancreatic residual infection and survival up to January 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers. Results:(1) Surgical situations: of the 20 patients who underwent surgery successfully, 14 patients underwent upper abdomen combined with left retroperitoneal approach, 1 patient underwent upper abdomen combined with right retroperitoneal approach, and the other 5 patients underwent upper abdomen combined with bilateral retroperitoneal approach. Fourteen of the 20 patients underwent additional surgery including 10 cases undergoing jejunostomy, 2 cases undergoing gastrostomy combined with jejunostomy, 1 case undergoing laparoscopic cholecystectomy combined with jejunostomy, and 1 case undergoing cholecystectomy. The operation time and volume of intraoperative blood loss of 20 patients were (228±41) minutes and 100 mL (range, 50-700 mL), respectively. (2) Postoperative situations: 20 patients began continuous perfused with 0.9% sodium chloride solution within the purulent cavity at postoperative day 2 (range, day 1-14). Six of the 20 patients had postoperative complications including 1 case with postoperative gastric fistula combined with intraperitoneal hemorrhage who underwent laparotomy hemostasis combined with gastrostomy at day 13 postoperatively, 1 case with postoperative duodenal fistula who underwent gastrointestinal anastomosis and jejunostomy at day 111 postoperatively, 1 case with postoperative retroperitoneal residual tissue necrosis and infection who underwent peripancreatic necrotic tissue debridement and drainage at day 11 postoperatively, 1 case with postoperative gallbladder fistula who underwent cholecystectomy at day 71 postoperatively, and 2 cases with postoperative pancreatic fistula who were cured with conservative treatment. The duration of hospital stay after 1st operation of the 20 patients were 42 days (range,20-178 days). (3) Follow-up: all 20 patients were followed up for 6.0 to 45.0 months, with a median follow-up time of 14.5 months. During the follow-up, 1 case developed secondary diabetes, and none of patient showed clinical manifestation such as fever, abdominal pain, abdominal distension and diarrhea. The peripancreatic residual tissue of all 20 patients absorbed well, and none of patient died.Conclusion:Bilateral route minimal-incision necrosectomy combined with continuous lavage is safe and feasible for the treatment of INP.

Objective:To prepare the site-specific biotinylation of enhanced green fluorescence protein with double biotin molecules using Avi-tag technology.Methods:The EGFP gene was prepared by PCR and cloned into pdi-Avitag resulting the vector pEGFP-( Avitag) 2.The fusion protein EGFP-( Avitag ) 2 was expressed in E.coli DH5αand purified by employing IMAC.The site-specific biotinylation was implemented by BirA enzyme in vitro, and then was identified by competitive ELISA and Western blot.Results:The recombinant prokaryotic expression vector pEGFP-(Avitag)2 was correctly constructed,and EGFP-(Avitag)2 fusion was successfully expressed in E.coli DH5α.The results of competitive ELISA and Western blot showed that the EGFP-( Avitag) 2 could be site-specific biotinylation with double biotin molecules based on Avi-tag technology.Conclusion: The site-specific biotinylation of EGFP with double biotin molecules is successfully prepared,and we anticipate that can be used for BAS to improve the sensitivity and specificity of immunosensors.

This paper reports a study regarding the verity of the hypocholestero-lemic effect of konjac-polysaccharide. The konnyaku powder (KP) used in this study was prepared and refined from the tubers of Amorphophallus ko-njac K. Koch and contained 84.8% of glucomannan. Male and female Spra-ue-Dawley rats aged 5 weeks were divided into 5 groups and fed on normal basal diet, hypercholesterolemic diet (control diet) and 3 test diets (i.e. KP was added to the control diet at a dosage of 2.5%, 5% or 10%) respe- ctively, for 12 weeks.The results obtained from this study showed that KP could markedly lower the level of the cholesterol in sera and livers of rats feeding hyper-cholesterolemic diets. At the end of the 4th week of the feeding experiment, the serum cholesterol level of the 5% and the 10% KP groups, and the liver cholesterol level of the 10% KP group were shown to be significantly lower than those of the control group. At the end of the 12th week, serum cholesterol levels of all the 3 KP groups were found to be lowered to the level of the normal group and so did the liver cholesterol level of the 10% KP group. The lipotropic (anticholesteatosis) effect of KP was also confirmed by the hepatic histopathological examination. Besides the hypocholest-eroletmic eftect, KP diets can also increase the bulk of stool. Finally, there were not any harmful effects on the absorption and utilization of Ca, Fe, Zn and Cu being found.

OBJECTIVETo evaluate the efficacy and safety of vardenafil on men with erectile dysfunction (ED) of various etiologies.

METHODSA total of 88 men with mild to severe erectile dysfunction were enrolled in the randomized, double-blind, placebo-controlled, fixed-dose trial of 12 weeks of treatment with either placebo or 5, 10 and 20 mg of vardenafil.

RESULTSThis study indicated that vardenafil dosages of 5, 10 and 20 mg were significantly superior to placebo for the treatment of ED, on the basis of the primary study endpoints of the EF domain score of the IIEF, diary-recorded success rates for penetration and maintenance of erection during the intercourse and the GAQ. Vardenafil was well tolerated. The incidence of adverse events was higher for vardenafil than for placebo.

CONCLUSIONOral vardenafil therapy has a high efficacy and a low incidence of adverse events for ED patients with mixed etiologies.

Administration, Oral ; Adult ; Aged ; Double-Blind Method ; Erectile Dysfunction ; drug therapy ; Humans ; Imidazoles ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Phosphodiesterase Inhibitors ; adverse effects ; therapeutic use ; Piperazines ; adverse effects ; therapeutic use ; Sulfones ; adverse effects ; therapeutic use ; Triazines ; adverse effects ; therapeutic use ; Vardenafil Dihydrochloride

This study aimed to observe the protective effect of momordicine I, a triterpenoid compound extracted from momordica charantia L., on isoproterenol (ISO)-induced hypertrophy in rat H9c2 cardiomyocytes and investigate its potential mechanism. Treatment with 10 μM ISO induced cardiomyocyte hypertrophy as evidenced by increased cell surface area and protein content as well as pronounced upregulation of fetal genes including atrial natriuretic peptide, β-myosin heavy chain, and α-skeletal actin; however, those responses were markedly attenuated by treatment with 12.5 μg/ml momordicine I. Transcriptome experiment results showed that there were 381 and 447 differentially expressed genes expressed in comparisons of model/control and momordicine I intervention/model, respectively. GO enrichment analysis suggested that the anti-cardiomyocyte hypertrophic effect of momordicine I may be mainly associated with the regulation of metabolic processes. Based on our transcriptome experiment results as well as literature reports, we selected glycerophospholipid metabolizing enzymes group VI phospholipase A 2 (PLA2G6) and diacylglycerol kinase ζ (DGK-ζ) as targets to further explore the potential mechanism through which momordicine I inhibited ISO-induced cardiomyocyte hypertrophy.Our results demonstrated that momordicine I inhibited ISO-induced upregulations of mRNA levels and protein expressions of PLA2G6 and DGK-ζ. Collectively, momordicine I alleviated ISO-induced cardiomyocyte hypertrophy, which may be related to its inhibition of the expression of glycerophospholipid metabolizing enzymes PLA2G6 and DGK-ζ.

Objective:To investigate the efficacy and safety of retroperitoneal soft tissue sarcoma resection combined with nephrectomy.Methods:The clinical data of 27 cases undergoing retroperitoneal soft tissue sarcoma resection combined with nephrectomy at the Gastrointestinal Tumor Center , Guangdong Provincial Hospital , Traditional Chinese Medicine from Jun 2017 to Aug 2022 were retrospectively analyzed for the indication of nephrectomy, postoperative progression of renal insufficiency and survival rate.Results:Twenty-six cases (96%) achieved R 0/R 1 resection and 1 case nderwent R 2 resection. Six cases underwent combined unilateral nephrectomy and 21 patients underwent combined multi-organ resection with a median number of resections of 4 (2,5). Postoperative pathology suggested that the combined resected kidney was positive for tumor infiltration in 17 cases. Five cases had Clavien-Dindo grade 3 or higher complications and no deaths occurred within 30 days after surgery. At the 90th day after surgery, 19 cases (70%) had decreased renal function ( Z=2.88, P=0.04), with a median decrease of -3.96 (-30.36, 0.31)ml·(min·1.73 m 2) -1, including 8 cases of preoperative Chronic Kidney Disease(CKD)1 stage progression (6 cases of CKD 2 stage, 2 cases of CKD 3 stage); 2 cases of CKD 2 stage progressed to CKD 3 stage; 1 case of preoperative CKD 3 stage progressed to CKD 4 stage. During the follow-up period of 3-38 months, no patient progressed to CKD 5 stage and no patient required dialysis treatment. Conclusion:Retroperitoneal soft tissue sarcoma resection combined with nephrectomy is safe and feasible while improving tumor radicality.

Objective This study aims to analyze and investigate the major factors that affecting stem cell clinical research,explore corresponding countermeasures,to further promote the clinical research and translational application of stem cells.Methods Comprehensive investigation were conducted to analyze and summarize the the most recent basic research of stem cells,standardization of stem cell preparation sources,human personnel,clinical research and supporting systems.Results The slow progress of stem cell clinical research could attribute to several factors,including sluggish frontier basic research of stem cells;lack of high-quality,clear-source,and controllable samples for stem cell research institutes,lack of standard operation codes,professional consensus,guidelines in multi-center clinical research and stem cell research,and unsound supporting system of stem cell clinical research.Conclusions Based on the analysis and summaries,combined with the practice and exploration of the author in past decade.Our recommendations include:focus on human major diseases,strive to make breakthroughs in key scientific problems of stem cell and translational research,establish a national stem cell resource bank,and conduct multi-center stem cell clinical research.Besides,increase the effort in personnel training in various related fields,establish a scientific system and mechanism for the supervision of stem cell clinical research,accelerate the clinical research and translation of stem cells,and finally contribute to the "Healthy China 2030" national strategy to provide better service to public.