To enhance the scientific research participation of the medical postgraduates, and pro-mote core competitiveness of the medical colleges, based on analysis of the necessity of patent application in medical colleges postgraduate, we constructed the teaching teams including the medical professors, patent engineers, graduate teaching manager, and science and technology managers. The problem-based learning (PBL) and case-based learning (CBL) as teaching methods were used in practice. The patent courses included the reference search and analysis, basic knowledge of patent law, and patent application training module were constructed; and the teaching effect were evaluated and optimized through the scores of the patent basic theory test, research output, and the training of the patent application. The patent course of the medi-cal postgraduates provides a reference for cultivating the compound talents have scientific research innova-tion and patent application capability.

OBJECTIVETo investigate lyophilization of SM-SLN.

METHODThe parameters of lyophilization process was optimized. In addition, the protective effect of various types and concentrations of cryoprotectants were tested by shape, colour and disparity.

RESULTThe mixture of 2% lactose and 2% glucose could better prevent nanoparticles from aggregating, the optimal lyophilization process was followed: precooled at -45 degrees C for 10 hr; primary drying at -25 degrees C for 5 hr; secondary drying at 10 degrees C for 3 hr; finally drying at 30 degrees C for 6 hr.

CONCLUSIONChanges in particle size distribution during lyophilization could be minimized by optimizing the parameters of the lyophilization process and adding supporting agent.

Drug Carriers ; chemistry ; Freeze Drying ; methods ; Glucose ; chemistry ; Lactose ; chemistry ; Lipids ; chemistry ; Milk Thistle ; chemistry ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Silymarin ; administration & dosage ; chemistry ; isolation & purification ; Technology, Pharmaceutical ; methods

Objective To study the sensitivity and specificity of a double monoclonal antibody sandwich enzyme-linked immunosorbent assay (DMcAbS-ELISA)for the detection of F1 antigen of Yersinia pestis (Y.pestis).Methods Viscera (viz.liver and spleen)specimens of infected mice with virulent Y.pestis and negative control mice were detected by bacteriological test,DMcAbS-ELISA and reverse indirect hemagglutination assay (RIHA) for the F1 antigen.Results The 225 control specimens were all negative tested by plague bacteriology testing,DMcAbS-ELISA and RIHA.A total of 308 plague-infected mouse organ specimens were tested,and the positive detection rate was 92.21％ (284/308),90.91％(280/308) and 89.61％ (276/308),respectively,with germiculture,DMcAbS-ELISA and RIHA,and the difference was not statistically significant(x2=5.65,P＞0.05).The coincidence rate of DMcAbS-ELISA and bacterial culture was 97.00％[(274+243)/533],Kappa =0.940;RIHA in line with the rate was 99.25％[(276+253)/533],Kappa =0.985.Authenticity comparison of F1 antigen detection in viscera specimens:sensitivity,specificity,positive predictive value,negative predictive value,adjusted agreement and Youden's index was 96.48％(274/284),97.59％(243/249),97.86％ (274/280),96.05 ％(243/253),96.99％[1/4×(274/280+274/284+243/253+243/249)]and 0.9407,respectively,for DMcAbS-ELISA and 96.13％(273/284),98.80％(246/249),98.91％(273/276),95.72％(246/257),97.39％[1/4×(273/276+273/284+246/257±246/249)]and 0.9492,respectively,for RIHA.The detection sensitivity of DMcAbS-ELISA and RIHA was 2.7×104 cfu/ml and 2.2×105 cfu/ml,for Y.pestis,respectively,and was 10 μg/L for F1 antigen.Conclusions DMcAbS-ELISA assay is a sensitive,specific,simple and fast method for detection of the F1 antigen,and it has a potential application value in rapid diagnosis of plague.

Objective To assess the quantitative relationship between the distribution of Himalayan marmot and its ecological environment,the terrain,the temperature and the precipitation,using remote sensing and geographic information system in Qinghai province.Methods The distribution of Himalayan marmot was located by Google Earth and ArcGIS software and by using field survey data provided by Chinese Center for Disease Control and Prevention.The corresponding ecological environment of marmot including terrain,temperature and precipitation were derived from the spatial information datasets.All results were processed according to the overlay and statistics analysis using ArcGIS software.Results Seventy-seven point twenty-seven percent(153/198) of Himalayan marmot were distributed in the area of elevation between 3000 and 4000 meters.The number of marmot reached the highest when the slope was between 0 and 17 degrees,and aspect range was between 91 and 270 degrees,180 degree was as south direction.During the period with the maximum temperature of the warmest month of 14.3-17.5 ℃,17.6-20.8 ℃ and 20.9-24.0 ℃,the distribution of marmot reached 95％(186/198) of the total area.Meanwhile,most of the marmot were presented in the area with average precipitation of 46-108 mm.Conclusions A quantitative analysis of appropriate ecological environment of Himalayan marmot in a large scope is carried uul successfully using remote sensing and geographic information system.The study indicates that spatial information technology has important applications in plague prevention and control.

Abstract: To explore the clinical characteristics, diagnosis and treatment of imported severe malaria and COVID-19 co-infection cases, and to provide scientific basis for epidemic prevention and control measures. The epidemiological characteristics, clinical manifestations, laboratory tests, treatment process and prognosis of 4 cases of severe malaria and COVID-19 co-infection with confirmed diagnosis were analyzed retrospectively. Four cases of severe malaria were African returnees of the same batch, male, aged 40-54 years old, with the same journey track. They all had African work and life history and acute onset. The main clinical manifestations were fever (4/4), chills (3/4), chills (3/4), nausea and vomiting (3/4), diarrhea (4/4), fatigue and anorexia (4/4). Two cases had headache and dizziness, confusion, muscle aches, two cases had cough, one cases had sputum, sore throat and runny urine. All 4 cases were confirmed by positive nucleic acid detection of the new coronavirus (2019-nCOV) in throat swabs. Plasmodium falciparum was found by microscopic examination of peripheral blood smears of all patients, and all of them were consistent with high altitude helminthiasis. All cases were accompanied by abnormal liver function and severe hypoproteinemia, two cases were hyperbilirubinemia, three cases were dyslipidemia, three cases were involved in abnormal tertiary hemogram with different degrees of elevation of procalcitonin, two cases were lactic acid poisoning, and one case was hypoglycemia. One case showed viral pneumonia on chest CT. All cases were treated individually according to the different conditions and were discharged after improvement, and were rechecked for 2019-nCOV nucleic acid and microscopic examination of blood smear negative for Plasmodium.During the global COVID-19 epidemic, the emergence of coinfection cases of con-infection of imported malaria parasites and severe acuterespiratory syndrome coronavirus 2 (SARS-CoV-2) makes the clinical diagnosis and treatment more complicated. It is important to establish the awareness of simultaneous prevention and diagnosis of COVID-19 and malaria for local prevention and control and early warning of severe cases, and timely and effective formulation of treatment plan to improve the comprehensive treatment efficiency.

Objective To further study the regulatory mechanisms of uncarboxylated osteocalcin on the glucose metabolism,rat uncarboxylated osteocalcin fusion protein was expressed,purified and identified.Methods The rat osteocalcin gene (BGLAP) was cloned into pSumo-Mut expression vector.The pSumo-BGLAP plasmid was obtained and identified.Through transformed in prokaryotic host bacterium and induced by isopropyl 3-D-thiogalactoside (IPTG) and 11 ℃ low temperature,the Sumo-BGLAP fusion protein was expressed.The purification and activity identification of the fusion protein was performed by Ni2+ affinity chromatography and insulin secretion experiments,respectively.Results The enzyme digestion and sequencing results of the plasmid showed that the recombinant plasmid pSumo-BGLAP was constructed successfully,which matched the target gene and protein.After the transformation,induction and purification,the high purity Sumo-BGLAP fusion protein was obtained.The data from insulin secretion experiments show that the fusion protein can promote insulin secretion under 16.7 mmol · L-1 glucose conditions [(37.64 ± 3.80)μU· L-1at 16.7 mmol · L-1glucose,(63.91 ±4.67) μU · L-1 at 16.7 mmol · L-1 glucose +0.03 ng· mL-1 Sumo-BGLAP,(68.47 ±5.83) μU · L-1 at 16.7 mmol · L-1 glucose +0.3 ng· mL-1 Sumo-BGLAP,P ＜ 0.01)].Conclusion The rat uncarboxylated osteocalcin fusion protein is successfully achieved and purified.The fusion protein with biological activity can be used to further study the regulatory mechanisms of uncarboxylated osteocalcin on the glucose metabolism.

The aim of the present study was to investigate the effect of a short-term (3-day) simulated microgravity with and without daily dorsoventral gravitation (-G(x)) for 1 h on myogenic tone and vasoconstrictor responsiveness of the middle cerebral artery and mesenteric third-order small artery in rats. The tail-suspension (SUS) model was used to simulate cardiovascular deconditioning due to microgravity. Daily restoring to normal standing (STD) posture for 1 h was adopted to provide -G(x) as the countermeasure. Segments of middle cerebral artery and mesenteric third-order small artery were isolated and cannulated. Vascular diameters in response to increased intraluminal pressure (from 20 mmHg to 120 mmHg, by 20 mmHg steps) of isolated arteries under no-flow conditions were recorded by a Pressure Myograph System in both physiologic salt solution (PSS) (active diameter, Da) and calcium-free PSS (passive diameter, Dp). The myogenic tone was calculated by (Dp-Da)/Dpx100%. Vasoconstrictor responsiveness of the isolated middle cerebral artery to serotonin and that of small mesenteric artery to phenylephrine were assessed in the PSS under an intraluminal pressure of 40 mmHg. The results showed that SUS induced an enhancement of the myogenic tone and vasoconstrictor responsiveness in the isolated middle cerebral artery but a depression of those in the small mesenteric artery. Daily STD for 1 h prevented the depression of myogenic tone and vasoconstrictor responsiveness in the small mesenteric artery, but did not prevent the functional enhancement in the middle cerebral artery. These data suggest that a short-term simulated microgravity may result in different alterations in the function of the cerebral artery and the resistance vessel in the hind-body. Moreover, only the decrease of function in these resistance vessels, not in the cerebral arteries, can be prevented by such a countermeasure of daily STD for 1 h.
Animals ; Cerebral Arteries ; pathology ; Hindlimb Suspension ; Mesenteric Arteries ; pathology ; Pressure ; Rats ; Serotonin ; pharmacology ; Vascular Resistance ; Vasoconstriction ; Weightlessness Simulation

AIM To establish a UFLC-QTRAP-MS/MS method for the simultaneous content determination of phenylalanine,isoleucine,methionine,valine,proline,tyrosine,aspartic acid,histidine,arginine,lysine and glutamic acid in Gynostemma pentaphyllum (Thunb.) Makino.METHODS The analysis of aqueous extract of G.pentaphyllum was performed on a Waters XBridge Amide column (2.1 mm × 100 mm,3.5 μm),with the mobile phase comprising of water-acetonitrile (containing 0.2％ formic acid) flowing at 0.6 mL/min in a gradient elution manner.RESULTS Eleven amino acids showed good linear relationships within their own ranges (r ＞0.998 9),whose average recoveries were 90.74％-103.05％ with the RSDs of 0.61％-5.00％.CONCLU-SION This accurate,stable and reproducible method can be used for the quality control of G.pentaphyllum.

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P

Objective To establish a method for detection of the human papi 11 omavirus(HPV)6b E7-specific antibodies in serum and cervical secretion from patients with condyloma acuminatum(CA).Methods A full-length HPV 6b E7 gene was amplified by PCR from the CA tissue to construct the recombinant plasmid pET32a(+)/HPV 6b E7.The expression of prokaryotic protein was analyzed by SDS-PAGE and Western blot,then purified with Ni-NTA Agarose affinity column and used as an diagnostic antigen for establishing indirect ELISA method,to detect specific serum IgG and specific cervical secretion slgA from 56 CA patients,81 healthy control.Sera from 43 cervical cancer was served as control.HPV 6b DNA from 56 CA patients was identified by PCR.Results Data showed that the nucleotide homology of cloned sequence was 99.5%,compared to the standard sequences of HPV 6b E7 gene(GenBank accession number:NC001355).A high level expression of E7 fusion protein was obtained in prokaryotic expression system(40 μg/ml).Based on HPV 6b E7 fusion protein being used as coating antigen,results from ELISA showed that the absorbance rates(A)of serum IgG from CA,cervix cancer and healthy control groups were 1.82±0.48,1.36 ± 0.39 and 1.39 ± 0.27,respectively.The level of IgG antibody in the serum of CA group was significantly higher than that in cervix cancer group and healthy control(P＜0.05).The A values of cervical secretion sIgA in CA and healthy control groups were 0.63 ± 0.26 and 0.53 ± 0.06,respectively,while the level of sIgA antibody in the cervical secretion of CA group was also significantly higher than that in healthy controls(P＜0.05).The positive rate of HPV 6b E7 DNA in CA tissue was 78.6%(44/56)by PCR method.When compared the results measured by PCR,the HPV 6b E7-specific IgG and sIgA antibodies by ELISA used to detect the patients infected with HPV 6b infection,showed that the sensitivity rates were 68.2%(30/44)and 54.6%(24/44)respectively,and the specificity were all 100%(12/12).Conclusion Based on the serum and cervical secretion specific HPV 6b E7 antibodies from patients with CA to diagnose HPV 6b infection,results showed medium sensitivity and high specificity,and could further be used to investigate the epidemiological characteristics of HPV 6b infection.