Objective: Soy products may cause excessive intestinal gas because of soybean oligosaccharides . The effect of recombinant ?-galactosidase on eliminating mouse flatus was observed. Methods:The mouse model of flatulence was set up by ig raffinose and stachyose and the flatulence was investigated by measuring intestinal flatulent volume. Oligosaccharides were examined by TLC test and soybean protein was examined by SDS-PAGE. Results: Raffinose and stachyose can result in mouse flatus and recombinant ?-galactosidase can eliminate it without any effect on soybean protein. Conclusion: Recombinant ?-galactosidase can eliminate flatus, and be used as food additive.

Objective To reduce immunogenicity of porcine skin by removingα-Gal epitopes expressed in cell surface and extracellular matrix using recombinant α-galactosidase produced by Bacteroides fragilis.Methods The porcine skin was harvested from healthy 2-month-old pigs without any skin disorders before being sterilized by iodine and 75%alcohol, respectively.Enzymatic removal of α-Gal antigen was followed by washing with PBS.The α-Gal antigen in the prepared porcine skin was measured with immunofluorostaining of cryosections and the residual enzyme was measured with a double-antibody sandwich ELISA method.Enzymatic removal procedures were optimized by detecting residual enzyme and the effi-cacy ofα-Gal removal under different enzymatic and washing conditions.Results Efficient enzymatic and washing methods were established to removeα-Gal antigen.Theα-Gal removal efficacy was above 90% and residual enzyme was undetect-able (αprescribed minimum ofα-galactosidase detection with indirect ELISA was 1 ng/ml) .Conclusion It is feasible to efficiently removeα-Gal antigen under these enzymatic and washing conditions, and a method of producing low-immunoge-nicity pig skin dressing for burn is established.

Objective To establish a method of quantiying trace α-galactosidase from Bacteroides fragilis in enzymatic conversion of blood group B to O red blood cells ( B-ECO RBCs) .Methods BALB/c mice were immunized with purified recombinant B.fragilisα-galactosidase ( the purity>90%) to prepare monoclonal antibodies.The ascites were prepared using hybridoma cell lines stably secreting antibody and purified by HiTrap rProtein A column.The antibody titer and spe-cificity were detected by ELISA and Western blotting, respectively.Purified monoclonal antibody and rabbit polyclonal an-tibody were applied to detect residual enzyme in B-ECO RBCs and the washing solution was analyzed by indirect ELISA. Results A high titer and purity antibody was obtained.Western blotting showed that the antibody specifically reacted with B.fragilisα-galactosidase.Moreover, indirect ELISA was sensitive enough to detect the minimal amount of residualα-gal-actosidase at the concentration of 1 ng/ml.After four repeat washing cycles with 1∶4 ( v/v) phosphate-buffered saline, the amount of residual enzyme in B-ECO RBCs was less than 10 ng/ml.Conclusion An effective method of detecting the min-imal amount of residual α-galactosidase in blood conversion is established for safety evaluation of universal RBCs prepara-tion by enzymatic treatment.

Aim ToestablishthemethodofHighper-formance liquid chromatography ( HPLC ) for detecting plasma concentration of indazole compound DL0805-1 , a Rho kinase inhibitor, and to investigate its pharma-cokinetics in rats with intravenous injection. Methods ThedetectingsystemwasAgilent1200-DAD;chro-matographic column was Agilent TC-C18 ( 4. 6 mm × 250 mm, 5 μm); the ultraviolet detection wavelength was 235 nm; the column temperature was 35 ℃; the flow rate was 1 ml·min-1;the mobile phase was ace-tonitrile-0. 05% H3 PO4 gradient elute. Rat blood sam-ples were collected at different intervals after intrave-nous injection of a single dose of DL0805-1 , and the concentration of DL0805-1 in rat plasma were deter-mined by HPLC method for estimating pharmacokinetic parameters.Results Afterintravenousinjectionof DL0805-1 in rats, prototype and its metabolite were detected in plasma. T1/2 of DL0805-1=(2. 34 ± 1. 42) h, Cmax=(3. 51 ± 0. 44) mg·L-1, T1/2 of metabolite of DL0805-1 = ( 1. 27 ± 0. 45 ) h, Cmax = ( 3. 55 ± 0.22)mg·L-1.Conclusion Theseresultssuggest that DL0805-1 may be metabolized into another sub-stance in vivo and play biological functions. The meth-od is sensitive, simple, and accurate, and can be used for the determination of DL0805-1 in rat plasma and pharmacokinetic studies.

Objectives:To establish hybridomas secreting monoclonal antibodies(McAbs) against O6-methylguanine-DNA methyltransferase(MGMT) and to observe the relationship between MGMT expression and clinical responses to ACNU and BCNU in human brain tumors.Methods:The hybridomas were established by cell fusion.MGMT expression in 60 glioma specimens was detected by means of immunohistochemical assay.Results: Seven hybridomas secreting McAbs against MGMT were obtained.Thirty tumor specimens had no detectable or low level of MGMT expression(Mer-), while 30 specimens had high level of MGMT expression(Mer+). The Mer- patients showed more sensitive to ACNU and BCNU than the Mer+ patients.Conclusions: The high specific hybridomas secreting monoclonal antibodies(McAbs) against MGMT were established.The preliminary study indicated that MGMT negative tumors were sensitive to ACNU and BCNU, whereas MGMT positive ones were more resistant to nitrosourea drugs.

OBJECTIVETo explore the strategies which reduce the amount of xenoantigen Galalpha1,3Gal.

METHODSHuman alpha-galactosidase gene and alpha1,2-fucosyltransferase gene were transferred into cultured porcine vascular endothelial cells PEDSV.15 and human alpha-galactosidase transgenic mice were produced. The Galalpha1,3Gal on the cell surface and susceptibility of cells to human antibody-mediated lysis were analyzed.

RESULTSHuman alpha-galactosidase gene alone reduced 78% of Galalpha1,3Gal on PEDSV.15 cell surface while human alpha-galactosidase combined with alpha1,2-fucosyltransferase genes removed Galalpha1,3Gal completely. Decrease of Galalpha1,3Gal could reduce susceptibility of cells to human antibody-mediated lysis, especially during co-expression of alpha-galactosidase gene and alpha1,2-fucosyltransferase gene. RT-PCR indicated positive human alpha-galactosidase gene expression in all organs of positive human alpha-galactosidase transgenic F1 mice including heart, liver, kidney, lung, and spleen, the amount of Galalpha1,3Gal antigens on which was reduced largely. 58% of spleen cells from F1 mice were destroyed by complement-mediated lysis compared with 24% of those from normal mice.

CONCLUSIONSHuman alpha-galactosidase gene and alpha1,2-fucosyltransferase gene effectively reduce the expression of Galalpha1,3Gal antigens on endothelial cell surface and confers resistance to human serum-mediated cytolysis. The expression of human alpha-galactosidase in mice can also eliminate the Galalpha1,3Gal antigens in most tissues and decrease the susceptibility of spleen cells to human serum-mediated cytolysis.

Animals ; Antigens, Heterophile ; metabolism ; Cell Death ; Cells, Cultured ; Disaccharides ; metabolism ; Endothelial Cells ; metabolism ; Fucosyltransferases ; genetics ; metabolism ; Graft Rejection ; genetics ; Humans ; Mice ; Mice, Transgenic ; Spleen ; cytology ; Swine ; Transfection ; alpha-Galactosidase ; genetics ; metabolism

Objective ToanalyzethevalueoftheDCE-MRIparametersofReferenceRegionandExtendedToftsmodleinthedifferential diagnosisofhyper-cellularuterineleiomyoma.Methods 59patientswithuterineleiomyomaconfirmedbysurgeryandpathologywere enrolledinthestudyfromSeptember2015toSeptember2016,including28casesofclassicalleiomyoma,12casesofcellularleiomyoma and19casesofdegenerativeleiomyoma.AllpatientsunderwentDCE-MRIbeforesurgery.Thequantitativeperfusionparameters (transferconstant(Ktrans),extravascularextracellularspacevolumeratio(Ve),effluxrateconstant(Kep),andbloodplasmavolume ratio(Vp)weremeasuredviaReferenceRegionmodel(RR model)andExtendedToftsmodel(ET model)respectively.ROCcurve wasusedtoevaluatetheefficiencyofthequantitativeperfusionparametersindifferentialdiagnosisofhyper-cellularuterineleiomyoma. Pearsoncorrelationanalysiswasalsoappliedtoobservethecorrelationsamongallequivalentparameters.Results KtransandKepinRR modelandKtrans,Kep,VeandVpinET modelshowedstatisticalsignificancesamongthreetypesofuterineleiomyoma (P＜0.05), meanwhile,theAUCsofKtrans,KepandVpderivedfrom RR modelandKtrans,Kep,VeandVpcalculatedbyET modelindiagnosisof cellularuterineleiomyomawere0.835,0.752,0.706,0.956,0.871,0.656and0.754.TheKtransintheRR modelwascorrelatedwith KtransintheET model(r＝0.600,P＜0.001),KepintheET modelwaspositivecorrelatedwithKepandVpinRR model(r＝0.275, P＝0.035;r＝0.376,P＝0.003).Conclusion Thevalueof Ktrans ,Kep and Vp in RR and Ktrans ,Kep ,Ve and Vp in ET modelcan be useful inthedifferentiationofhyper-cellularleiomyoma,especiallythevaluesofKtransandKepinET modelhavehigherdiagnosticefficacy thanthoseinRR model.

Salvianolic acid A is a water-soluble component from Danshen, which is frequently used in traditional Chinese medicine. High performance liquid chromatography was often used to analyze content of salvianolic acid A. The yield of salvianolic acid A increased by the technological improvement of extraction and separation. Salvianolic acid A possessed multiple pharmacological activities, including antioxidants, myocardial ischemic protection, antithrombatic, neuroprotection, anti fibrosis, prevention of diabetes and complications. Recently, preliminary pharmacokinetics characteristics of salvianolic acid A were clarified. Based on the research literature and study work from author's laboratory, this review will focus on recent developments concerning the chemistry, pharmacology and pharmacokinetic of salvianolic acid A, and prospect further research.
Animals ; Biomedical Research ; Caffeic Acids ; chemistry ; isolation & purification ; pharmacology ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; Lactates ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Salvia miltiorrhiza ; chemistry

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the outbreak of coronavirus disease 2019 in Wuhan City, China. The SARS-CoV-2 is genetically similar to the coronavirus derived from bat. The SARS-CoV-2, the SARS-CoV and the Middle East respiratory syndrome coronavirus (MERS-CoV) all belong to beta coronavirus. Since the outbreak of the coronavirus disease 2019, effective antiviral drugs have become a hot issue in the world. Very little about SARS-CoV-2 is known and there is no precedent for treatment. The National Health Commission has repeatedly revised the diagnosis and treatment guide for the coronavirus disease 2019. The latest guide is "New Coronary Virus-Infected Pneumonia Diagnosis and Treatment Plan (Seventh Trial Version)"(short for Seventh Version of Diagnosis and Treatment Plan). But the use of antiviral drugs is still on trial and no rigorous clinical trials data is available. Hot anti-SARS-CoV-2 drugs include interferon α, ribavirin, lopinavir/ritonavir, chloroquine phosphate, abidol, as well as hydroxychloroquine sulfate and remdesivir. But the later 2 drugs aren't mentioned in the Seventh Version of Diagnosis and Treatment Plan.
Antiviral Agents ; therapeutic use ; Betacoronavirus ; China ; Coronavirus Infections ; drug therapy ; Humans ; Pandemics ; Pneumonia, Viral ; drug therapy ; Practice Guidelines as Topic

OBJECTIVES: Obstructive sleep apnea hypopnea syndrome (OSAHS) is a common disease that seriously affects health. Continuous positive airway pressure (CPAP) therapy is the preferred treatment for moderate-to-severe OSAHS patients. However, poor adherence to CPAP is a major obstacle in the treatment of OSAHS. Information-motivation-behavioral (IMB) skills, as a kind of mature technology to change the behavior, has been used in various health areas to improve treatment adherence. This study aims to explore the effects of the IMB skills intervention on CPAP adherence in OSAHS patients. METHODS: Patients who were primary diagnosed with moderate-to-severe OSAHS were randomly divided into the IMB group (n=62) and the control group (n=58). The patients in the IMB group received CPAP therapy and the IMB skills intervention for 4 weeks. The patients in the control group received CPAP therapy and a usual health care provided by a registered nurse. We collected the baseline data of the general information, including age, sex, body mass index (BMI), the Epworth Sleepiness Scale (ESS) score, the Hospital Anxiety and Depression Scale (HADS) score, and indicators about disease severity [apnea-hypopnea index (AHI), percentage of time with arterial oxygen saturation SaO₂<90% (T90), average SaO₂, lowest SaO₂, arousal index]. After CPAP titration, we collected CPAP therapy-relevant parameters (optimal pressure, maximum leakage, average leakage, 95% leakage, and residual AHI), score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. After 4 weeks treatment, we collected the ESS score, HADS score, CPAP therapy-relevant parameters, effective CPAP therapy time per night, CPAP therapy days within 4 weeks, CPAP adherence rate, score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. Visual analog scale (VAS) of 0-5 was used to evaluate the satisfaction and acceptance of IMB intervention measures in the IMB group. RESULTS: There were no significant differences in the baseline level of demographic parameters, ESS score, HADS score, disease severity, and CPAP therapy related parameters between the IMB group and the control group (all P>0.05). There were no significant differences in score of willingness to continue CPAP therapy, as well as score of satisfaction and acceptance of CPAP therapy after CPAP titration between the IMB group and the control group (both P>0.05). After 4 weeks treatment, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group were significantly decreased, while the score of satisfaction and acceptance of CPAP therapy and willingness to continue CPAP therapy of the IMB group were significantly increased (all P<0.05); while the above indexes in the control group were not different before and after 4 weeks treatment (all P<0.05). Compared with the control group, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group after 4 weeks treatment were significantly lower (all P<0.05); the effective CPAP therapy time, CPAP therapy days within 4 weeks, score of satisfaction and acceptance of CPAP therapy, score of willingness to continue CPAP therapy of the IMB group were significantly higher (all P<0.05). The rate of CPAP therapy adherence in 4 weeks of the IMB group was significantly higher than that of the control group (90.3% vs 62.1%, P<0.05). The VAS of overall satisfaction with IMB skills intervention measures was 4.46±0.35. CONCLUSIONS IMB skills intervention measures can effectively improve the adherence of CPAP therapy in OSAHS patients, and is suitable for clinical promotion.
Continuous Positive Airway Pressure ; Humans ; Motivation ; Oximetry ; Patient Compliance ; Sleep Apnea, Obstructive/diagnosis* ; Syndrome