Objective To explore the feasibility of the three-port laparoscopic surgery through single umbilical incision with urological desease. Methods Thirty-two patients (10 males and 22 females) were taken the laparoscopic surgery using three ports through single incision. Including varicocele 7 cases, simple kidney cyst 12 cases, double kidney cyst 1 case, polycystic kidney 1 case, left adrenal tumor 3 cases, right adrenal tumor 1 cases, left upper ureteral calculi 1 cases, giant hydronephrosis 1 case and atrophic kidney 4 cases. The surgery procedures were including make a 1.0-3.0 cm long incision in the navel, followed by inserting three 10 mm or 5 mm trocars in the incision for observation and operation. Conventional laparoscopic techniques were used to complete the urological surgery. Results The operation time of varicoeele ligation was 10--20 rain, mean 15 min, no intraoperative bleeding. The operation time of renal cysts was 30-53 min, mean 40 rain, no intraoperative bleeding. The operation time of resection of adrenal tumor was 57--120 min, mean 68 rain, intraoperative bleeding was 20-60 ml, mean 30 ml. The operation time of ureterolithotomy was 86 min, intraoperative bleeding was 50 ml. The operation time of nephrectomy was 45-135 min, mean 65 min, intraoperative bleeding was 90-150 ml, mean 110 ml. Length of stay 3-8 days, average 5.5 days.With average follow-up time 2 months, all cases were fully recovered without complication and no visible scar in the abdominal region. Conclusion The laparoscopic surgery using three ports through single incision is safe and effective in selected urological surgery.

AIM: Screening of phage antibodies against HBsAg from the constructed phage display library and sequencing of the positive clones.METHODS: RNAs were extracted from blood lymphocytes of a HBsAg-immunized volunteer. Then cDNAs were synthesized by RT-PCR. To construct the phage antibody display library, cDNAs of Fd fragments and ? chains of the antibodies were amplified through PCR and were inserted into vector pComb3H. Three rounds panning against coated HBsAg showed the specific enrichment of phage antibody. Positive clones were selected and sequenced.RESULTS: A phage antibody display library was constructed after amplifing of Fd fragments and ? chains of IgG1. Three Fab antibodies from the library were selected and sequenced. The result of sequencing showed that two of the three antibodies had the same Fd fragment and all of the three had the same light chain. CONCLUSION: The phage antibody display library was constructed successfully and three specific antibodies (Fab fragments) have been obtained from it.

Objective To analyze the computed tomography (CT)characteristics of splenic lymphangioma and the association be-tween CT findings and pathological results.Methods The CT characteristics and pathological findings of 9 patients with splenic lym-phangioma were retrospectively analyzed.Results There were 8 cavernous lymphangioma and 1 cystic lymphangioma.Of the 9 ca-ses,it revealed that the percentage of blood vessel elements ranging from 5% to 50% via microscope.CT found 2 cases were with single lymphangioma,3 with multiple lesions,exhibiting as round or round-like mass;4 cases were found with diffuse lymphangio-ma and different size of cystic masses distributed across spleen.Five of the 9 cases who were with single or multiple lymphangioma showed circular and thin line-like cyst wall,while the remaining 4 cases showed latticed cyst wall in CT characteristics.The content in the cysts were with uneven density in all the 9 cases with CT value ranging from 10 to 40 HU,3 of which combined with sand-like calcification.Enhancement scanning found two characteristics:(a)cyst wall and separation were mildly enhanced,especially in the delayed phase;(b)the content in the cyst presents anomalous small patchy and mild enhancement.The enhancement of the content in the cyst did not change as the increasing of blood vessel composition.Conclusion CT examination will help the diagnosis of splenic lymphangioma and is of significance in informing clinical treatment.

AIM: To purify and refold the inclusion body of a human anti-HBsAg scFv with a 6?His tag, and to determine the affinity constant of the purified recombinant product.METHODS: Solubilizing in buffers containing urea or guanidine hydrochloride (GuHCl), the inclusion body was purified by IMAC, and then refolded by dialysis against urea or GuHCl, at the same time, Ni 2+ charged chelate column was utilized for in situ refolding. The affinity constant of the refolded scFv, polished by immune-affinity chromatography, was determined by non-competitive ELISA. RESULTS: The refolded scFv with highest specific bioactivity was produced by dialysis against GuHCl. Under this condition, the recovery of target protein reached (61.08?1 45)%. The affinity constant of the polished scFv was confirmed to be(2.30?0.32) ?10 7 L/mol. CONCLUSION: The inclusion body studied in this paper can be refolded efficiently under optimal dialysis condition in vitro . The antigen-binding property of this recombinant scFv is not affected by the purification tag fused to the N terminal of the protein.

AIM:To study the high-level HBV replication transgenic mice for evaluation of drugs treating hepatitis B virus.METHODS:The HBV transgenic mice were treated respectively with lamivudine,large dose recombinant hepatitis B protein vaccine,?-1b interferon,siRNA to evaluate their pharmacodynamics and mechanism of action.RESULTS:HBV DNA titre was reduced significantly in transgenic mice which were treated with lamivudine(100 mg?kg-1?d-1),recombinant hepatitis B protein vaccine(HBsAg 6 ?g/mouse),?-1b interferon(50 ?g /mouse),respectively.Recombinant hepatitis B protein vaccine and ?-1b interferon promoted the level of IL-2 and IFN-? and increased the Elispot number of spleen cells secreting IFN-? in the treated transgenic mice.HBV transgenic mice were treated with RNAi expression vector pU6-siHBV against HBV through vena caudalis by hydrodynamics technique.Five days later,the level of serum HBsAg was reduced by 56.7% and the inhibition lasted at least 14 days.The HbcAg(+)cells were decreased obviously by immunohistochemistry detection in liver tissue,but the RNAi did not reduce the serum HBV DNA titre.CONCLUSION:These inbreeding high-level HBV replication transgenic mice are reliable and feasible for evaluating the anti-HBV drugs and have its economical and convenient superiority.

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Drug Synergism ; Escherichia coli ; genetics ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; Umbilical Cord ; cytology

Objective: To prepare strains of influenza A (H7N9) pseudovirus derived from different districts of China for vaccine efficacy evaluation. Methods: Phylogenetic tree was built based on hemagglutinin (HA) amino acid sequence analyses from 29 influenza A (H7N9) virus strains and 6 influenza A (H7N9) virus strains with HA determinants variation were selected. 293FT cells were co-transfected with plasmid pNL4-3-Luc.R-E-, pVRC-HA and pVRC-NA with codon-optimized hemagglutinin (HA) and neuraminidase (NA) derived from the six influenza A (H7N9) virus strains, respectively. Transmission electron microscopy assay and Western blot analysis were performed to demonstrate morphology and specificity of these particles, luciferase activity assay and hemagglutinin titers detection were used to determine their infectivity and hemagglutinin activity. And finally, pseudovirus-based neutralization assays were evaluated with HA immunized mice serum. Results: Six influenza A (H7N9) peseudovirus particles derived from different districts of China were selected and prepared. All of the particles bearing HA and NA were characterized with classic influenza virus morphology, with TCID₅₀ titer ranged from 10⁴TCID₅₀/50 μl to 10⁵TCID₅₀/50 μl and with hemagglutinin activity ranged from 64 to 512. Neutralization efficacies on influenza A/Shanghai/1/2013(H7N9) HA vaccine serum against 100TCID₅₀ dose of these pseudovirus particles indicated their potential application in the vaccine cross-protective evaluation in future. Conclusions Six influenza A (H7N9) pseudovirus derived from different districts of China with potential antigenic variation on HA were constructed successfully, established foundation for their further application in vaccine cross-reactive efficacy evaluation.

Objective: To investigate the indication and midterm outcomes of surgical treatment of traumatic tricuspid insufficiency. Methods: Totally 19 patients with traumatic tricuspid insufficiency who underwent surgical treatment at Department of Cardiac Surgery, Beijing Anzhen Hospital, Capital Medical University from January 2002 to January 2018 were included in this retrospective study. There were 12 male and 7 female patients, aged (43.1±12.9) years (range: 17-68 years). The main causes of traumatic tricuspid insufficiency included blunt chest trauma following high-speed vehicle accidents (17 patients) and high-fall trauma (2 patients). The preoperative New York Heart Association functional class was class Ⅱ in 5 patients, class Ⅲ in 12 patients, and class Ⅳ in 2 patients. The mechanism of tricuspid insufficiency included anterior chordal rupture in 9 patients, anterior papillary muscle rupture in 3 patients, anterior and posterior chordal or papillary muscle rupture in 4 patients, laceration of leaflet combined with chordal rupture in 2 patients and infection combined with anterior papillary muscle rupture in 1 patient. Anular dilation and enlargement of the right ventricle were observed in all the patients. Paired t test was used to evaluate the echocardiogratic results at preoperation, postoperation and follow-up. Independent sample rank sum test was used to evaluate the intervals between trauma and surgery in tricuspid valve repair group and tricuspid valve replacement group. Results: Tricuspid valve repair was successful in 8 patients, and 11 patients underwent valve replacement. Among the patients who underwent valve replacement, 6 patients received mechanical valve and 5 received bioprosthetic valve. The interval from trauma to surgery of the valve repair group and valve replacement group were 8.5(10.0) months (range: 0.1-13.0 months) and 72.0 (108.0) months (range: 2.0-228.0 months), respectively. Concomitant procedures included debridement in scalp trauma (1 patient), internal fixation of femoral fracture (1 patient). One patient died from liver failure 10 days after operation and the remaining patients survived. Eighteen patients were followed up for (94±50) months, 15 patients were in New York Heart Association functional class Ⅰ and 3 patients in class Ⅱ. One patient received redo-tricuspid valve replacement because of mechanical valve failure at the 11 years of follow-up. Conclusions The midterm outcomes of surgical treatment of severe traumatic tricuspid insufficiency were satisfactory. Early diagnosis and surgical invention were recommended to achieve successful valve repair.

Objective: This study aims to explore the relationships of family functioning, general well-being, and exercise with psychological distress.Furthermore, we investigated the special roles of general well-being and exercise on the association between family functioningand psychological distress. Methods: Of 769 end-stage renal disease (ESRD) patients participated in the cross-sectional study which consisted of the 12-item GeneralHealth Questionnaire (GHQ-12), the Family APGAR Scales, and the General Well-Being Schedule. The collected data were analyzedusing multiple linear regression analysis and path analysis. Results: The prevalence of psychological distress was 72.3%. Family functioning, general well-being and exercise were associated factorsof psychological distress (p<0.05). The indirect effect of family functioning on psychological distress was partially mediated by generalwell-being (Effect=-0.08, 95% CI=-0.11, -0.04). In addition, the effect of family functioning on general well-being was moderated byexercise (Index=-0.092, SE=0.033, 95% CI=-0.159, -0.029). Conclusion The prevalence of psychological distress among ESRD patients was high. Family functioning, general well-being and exercisewere associated with psychological distress. Family functioning could affect psychological distress partially by affecting general wellbeing.Furthermore, exercise had a significant moderating effect on the relationship between family functioning and general well-being.Psychiatry Investig 2020;17(4):356-365

Membrane creates the functions of protection, supporting, dispersion and separation. More functions can be designed by modifying membrane surface and grafting/loading selective ligands or catalysts on the membrane, thus membrane technology has been widely applied in biological detection, and its application approaches becomes diverse. Rational design of functional membranes can meet the demands in different steps of biological detection process, including sample pretreatment, preparation, response and sensing. This review summarized the functionalization methods of filtration membranes, applications of membrane technology in sample preparation and detection process, as well as the research on the integration of functional membranes. By revisiting the research progress on functional membrane design, preparation and applications for biological detection, it is expected to take better advantage of membrane materials structure and performance for constructing efficient and stable detection platform, which is more "adapted" to the detection environment.
Membranes, Artificial