Construction and application effect of teaching resources of human physiology course from the perspective of demand analysis theory / 中华医学教育探索杂志

Objective:Based on the theory of demand analysis theory, to build teaching resources of human physiology course and explore its application effect.Methods:The three-year nursing junior college students of Batch 2018 were selected as the research objects. According to the principle of randomization, the students were divided into the observation group (97 cases) and control group (99 cases). Students in the observation group used integrated teaching resources from the perspective of demand analysis, while students in the control group used traditional curriculum resources. After the end of the course, the differences between the two groups of students' theoretical and practical assessment results and teaching experience were compared SPSS 20.0 was used for t test. Results:The results of theoretical [(73.67±8.76) vs. (70.48±8.36)] and practical skills [(84.41±4.83) vs. (83.63±4.27)] in the observation group were significantly different from those in the control group ( P<0.05). The overall scores of 9 teaching effect sub-items, such as learning awareness of inquiry, independent learning ability and critical thinking ability, were significantly higher than those of the control group ( P<0.01). Conclusion:The construction of human physiology teaching resources based on demand analysis theory can improve the effectiveness and pertinence of resource construction, and the effective use of resources in teaching can significantly improve the teaching effect, stimulate students' subjective initiative, cultivate their comprehensive ability and improve classroom efficiency.

Analgesic effect of α-cobratoxin on mice and its effect on protein kinase A activity of spinal dorsal root ganglion in mice / 中华行为医学与脑科学杂志

Objective:To study the analgesic effect of α-cobratoxin (α-CbTX) on mice and its effect on protein kinase A (PKA) activity of spinal dorsal root ganglion (DRG) in mice.Methods:Healthy male ICR mice( n=102) were randomly divided into low-, medium-, and high-dose α-CbTX groups (1 mg/kg, 3 mg/kg, 9 mg/kg respectively, gavage, n=21), solvent control group (equivalent volume of 0.9% normal saline, gavage, n=21), morphine positive control group (3 mg/kg, intraperitoneal injection, n=6)or aspirin positive control group(300 mg/kg, gavage, n=12). The analgesic effect of α-CbTX was evaluated by hot plate test, acetic acid twisting test and formalin foot licking test. Formalin plantar injection was used to induce pain and then the L4-L6 DRG was taken 30 minutes later. The expression of PKA C-α in L4-L6 DRG of mice were detected by Western blot.SPSS 16.0 software was used for statistical analysis. Repeated measurement ANOVA was used to evaluate the hot plate experimental data, and one-way ANOVA was used for other experimental data. LSD- t test was used for further pairwise comparison. Results:In the hot plate test, the interaction between group and time of mice paw licking latency was significant ( F=8.902, P<0.05). At 0.5 h after administration, the paw licking latencies of α-CbTX medium-dose group ((11.83±1.47)s)and α-CbTX high-dose group (( 14.33±12.1)s) were both longer than that of solvent control group((8.17±0.75) s) ( t=4.461, 7.053, both P<0.05). The efficacy of α-CbTX medium dose group lasted until 1.5 h after administration (all P<0.05), and that of α-CbTX high dose group lasted until 2 h after administration(all P<0.05). In the acetic acid writhing test, the writhing times in the low-, medium- and high-dose α-CbTX group((34.50±3.62) times, (26.17±2.40) times, (13.83±3.76) times)) were significantly lower than that in solvent control group ((42.50±4.59) times) ( t=3.938, 8.040, 14.112, all P<0.05). In the period of the formalin test phase Ⅱ, the total licking time of α-CbTX low-, medium- and high-dose groups ((71.17±6.46) s), (54.67±6.41) s, (40.50±3.89)s) were significantly shorter than that of the solvent control group ((98.67±11.50) s)( t=6.950, 11.120, 14.700, all P<0.05). In the Western blot experiment, compared with solvent control group (0.22±0.01), the levels of PKA C-α in the DRG of mice in low-, medium- and high-dose α-CbTX groups ((0.31±0.02), (0.41±0.03), (0.44±0.02)) were up-regulated ( t=3.140, 6.471, 7.492, all P<0.05). Conclusion:α-CbTX has obvious analgesic effect, and its analgesic mechanism may be related to the activation of PKA.

Advances in biomolecular machine: methane monooxygenases / 生物工程学报

Methane monooxygenases (MMO), regarded as "an amazing biomolecular machine", catalyze the oxidation of methane to methanol under aerobic conditions. MMO catalyze the oxidation of methane elaborately, which is a novel way to catalyze methane to methanol. Furthermore, MMO can inspire the biomolecular machine design. In this review, we introduced MMO including structure, gene and catalytic mechanism. The history and the taxonomy of MMO were also introduced.

Comparative study of the titanium and titanium alloy implant electrolytic etching surface / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This preliminary study aims to investigate the effects of titanium and titanium alloy micro-nano-dimensional topography on the biological behavior of osteoblasts in vitro.</p><p><b>METHODS</b>Electrolytic etching (EE) method was used to produce micro-nano dimensional titanium surfaces. The surfaces were observed to determine their effects on the adhesion, proliferation, cell morphology, and alkaline phosphatase (ALP) activity of osteoblasts.</p><p><b>RESULTS</b>The surfaces of the titanium and titanium alloy groups exhibited higher adhesion and proliferation of osteoblasts than those of the mechanical group. The titanium surface was covered with a group of cells, a large number of filopodia, and functional particles. The ALP activity of the titanium group was significantly higher than that of the titanium alloy and mechanical groups.</p><p><b>CONCLUSION</b>EE method in pure titanium and titanium alloy surfaces result in bowl-like nests and nanostructures of different diameters and depths. The diameters of the pure titanium and titanium alloy surfaces range from 30 to 50 μm and 5 to 8 μm, respectively. The former is more conducive to promote the proliferation and differentiation of cells.</p>

The measurement of affinity of site-specific labeled annexin V with ~(99m)Tc by calcium titration / 基础医学与临床

Objective To derermine the affinity of site-specific labeled annexin V with ~(99m)Tc to phosphatidylserine (PS)exposed erythrocytes.Methods The annexin V fused with a metal chelating binding site was obtained from Pichia Pastoris culture and methanol induction expression.The annexin V was purified from the culture supernatant crude product by uhrafihration.The annexin V was conjugated with ~(99m)Tc site-specifically through sodium glucoheptonic acid and SnCl_2.The radioactive annexin V was added to determine its affinity to PS exposed erythrocytes.Results The calcium concentration at which half of the protein is bound to cells(EC50)is changed with varying ratio of protein to cells.The affinity was determined at a low protein/cell ratio as 33.4.Conclusion The annexin V recombinantly expressed in Pichia Pastoris shows high affinity to PS exposed erythrocytes.

Association between WNK4 polymorphism and essential hypertension in Kazak population in China / 中华老年医学杂志

Objective To identify the genetic variants of WNK4(with no K=lysine kinase)gene in Kazak population in Xinjiang province,to determine whether the WNK4 gene intron 10 polymorphism iS associated with essential hypertension(EH)and to investigate the distribution of genotype and allele frequencies of WNK4 gene. Methods One hundred and ninety-one patients with EH and 173 normal blood pressure controls were included in the study to assess the contribution of polymorphism of WNK4.Direct DNA sequencing was performed to identify the single nucleotide polymorphism(SNP)in 16 SUbjects with EH and 16 subjects of normotension(NT).Then the polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)method was used for the detection of WNK4 genotype. Results One SNP located in the 10 intron of WNK4 (1156666 base of chromosome 17)was found in the Kazak population.The genotypes of the variants were found to be in Hardy-Weinberg equilibrium.GG,AG,AA genotypes were 88.0%,11.0 oA,1.0%in the EH group and 91.9%,8.1%,0%in the NT group,respectively.The frequencies of genotype and allele in EH group were not significantly different from NT group in Kazak population.Conclusions It suggests that the intron 10 polymorphism of WNK4 gene might be not associated with hypertension in Kazak population.

The packaging and purification of recombinant adeno-associated virus loaded with anti-amyloid ? peptide single-chain antibody gene / 基础医学与临床

Objective To produce and purify recombinant adeno-associated virus(AAV) loaded with anti-amyloid ? peptide single-chain antibody gene for gene therapy of Alzheimer's disease.Methods The plasmid pSNAV2.0-Abeta-scFv was used to transform BHK-21 cell by Lipofectamine 2000 for stable package cell line establishment.The helper virus HSV1-rc/?UL2 was used to transfect package cell for anti-amyloid ? peptide single-chain antibody gene loaded recombinant adeno-associated virus production.The chloroform-PEG/NaCl-chloroform extraction and ion-exchange chromatography were employed for recombinant AAV purification.SDS-PAGE and PCR amplification were adopted for purified virus identification.The final virus physical titer was determined by digoxin-labeled DNA probes.The effect of gene therapy was tested with transgenic mice by water maze test.Results The purity of recombinant adeno-associated virus with our target gene reach up to 98% after stable cell package and serial purification.The physical titer of the final virus was 1?1012vg/mL.The latency of treated mice in water maze test were reduced significantly.Conclusion The adeno-associated virus carrying anti-amyloid? peptide single-chain antibody gene was produced by HSV1system and purified.The animal behavior test demonstrated the recombinant AAV waseffective in the gene therapy of Alzheimer's disease.

Expression of Human Single-chain Antibody Against A?40 in Pichia Pastoris / 医学研究杂志

Objective To express human single-chain antibody against amyloid?peptide 40 in pichia pastoris for passive immuniza- tion to Alzheimer's disease patients.Methods Human single-chain antibody gene from a phage display library was mutated to obliterate BamHI site and cloned into pAO815 plasmid for pT-scFvA?40 construction which was identified by PCR amplification and endonuclease digestion.Vector pT-scFvA?40 was cut by EcoRI and NotI endonucleases and the antibody gene was cloned into pPICgK plasmid to con- struct pPIC9K-scFvA?40 expression vector which was confirmed by gene sequencing.Linearized pPIC9K-scFvA?40 was used to trans- form pichia pastoris GS115 and the recombinant was induced by 0.5% methanol to express human single - chain antibody against amyloid?peptide 40.Results Gene sequencing confirmed pPIC9K-scFvA?40 orientation.Recombinants were obtained by linearized pPICgK - scFvA?40 transformation.After inducing with 0.5% methanol,the recombinant secreted proteins with 33 kD size.Conclusions Expression vector pPIC9K-scFvA?40 was constructed successfully.Human single-chain antibody against amyloid?peptide 40 was recombinantly expressed in pichia pastoris.

The Preparation Investigation on Human Annexin V Dimmer(diannexin) / 医学研究杂志

Objective To prepare human annexin V dimmer and make chance for its pharmacological research.Methods Pichia pastoris transformed by human annexin V gene was used in this investigation.The engineered yeast was cultured in BMGY media and induced with methanol media BMMY for secreting expression of human annexin V.The final BMMY media supernatant was adopted for preparation of human annexin V dimmer by ammonium sulfate precipitation,molecular sieve separation and ultra-filtration concentration.The purification process was monitored by SDS-PAGE and coomassie brilliant blue staining.Results Human annexin V was obtained after two days induction of pichia pastoris in BMMY media.The human annexin V dimmer was successfully prepared from culture supernant by serial ammonium sulfate precipitation,molecular sieve separation and ultra-filtration concentration.Conclusion We prepare the human annexin V dimmer(diannexin V)successfully.

The Conjugation of Fluorescein Isothiocyanate to Annexin V and its Application / 医学研究杂志

Objective To prepare a novel annexin V and conjugate it with fluorescein isothiocyanate and test their binding ability to phosphatidylserine(PS)exposed erythrocytes.Methods The annexin V fused with an metal chelating binding site was obtained from pichia pastoris culturing and methanol induction expression.The annexin V was purified from the culture supernatant crude product by ion-exchange chromatography and ammonium sulfate precipitation.The annexin V was conjugated with fluorescein isothiocyanate in boric buffer and the conjugate annexin V-FITC was purified by ion-exchange chromatography.Sheep red blood cells were treated with Glutaraldehyde to expose membrane PS.The annexin V-FITC binding to PS exposed erythrocytes was investigated by fluorescent microscopy.Results The novel annexin V was purified and concentrated from pichia pastoris culture by ion-exchange chromatography and ammonium sulfate precipitation.After conjugating with FITC,the annexin V was found to bind PS exposed erythrocytes by fluorescent microscopy.Conclusion The annexin V expressed recombinantly in pichia pastoris retains the binding ability to PS exposed erythrocytes and is applicable to apoptosis detection in vitro.

Echocardiographic guidance of transcatheter closure of vent ricular septal defects using the Amplatzer device / 中华超声影像学杂志

Objective To evaluate the applied values of echocadiography in closing membranous ventricular septal defects (VSD) with the Amplatzer devices. Methods The VSD were closed by transcatheter Amplatzer closure devices in nine patients. The patients′ age ranged between 15-45 years old,mean (26?10) years old. The VSD diameter was measured from 3 to 15 mm. Seven patients were guided by transthoracic echocardiography and 2 patients by transesophageal echocardiography. Results All VSD were successfully closed by the Amplatzer devices, without complications. The device size ranged from 6 to 18 mm. No residual shunt and complication were detected by the echocadiography in intraoperation and 3 or 4 days after operation. Conclusions Echocardiography plays an important role in closing VSD with Amplatzer device, not only in intraoperative guidance and monitor, but also in post-operative follow up study.

Correlation of TCM Syndrome Patterns of Chronic Pharyngitis with Related Factors / 广州中医药大学学报

Objective To explore the correlation of TCM syndrome patterns of chronic pharyngitis(CP)with etiological factors.Methods A questionnaire investigation was carried out in 102 CP patients,of which 30 were differentiated as lung-kidney yin deficiency,30 as spleen-stomach deficiency,30 as phlegm coagulation and blood stasis,and 12 as spleen-kidney yang deficiency.The dietary hobbies,living and working environment,systemic diseases and living habits of the patients were recorded.Meanwhile,parameters of the pharyngeal microorganism and immunology were detected.Results The results of nonparametric test showed significant difference between the age and the mean disease course of the four syndrome patterns(P0.05). There existed significant difference between indulging in spicy food,indulging in cold and raw food,and staying up at night in the four syndromes (P0.05).The distribution of the four syndromes was different in the population with medical history of tonsillitis,dental caries,chronic cough,chronic gastritis and chronic colitis(P0.05).Conclusion The syndrome patterns of chronic pharyngitis are correlated with age,disease course,dietary habits of indulging in spicy food and cold-raw food,living habit of staying up at night,systemic disease or disease of organs around the throat,but are not correlated with sex,household registration,job,working and living environment,indulging in frying food,hobbies of smoking and drinking,living habits,emotional factors,throat swab culture results and immune parameters.

Establishment of Guinea Pig Models of Allergic Rhinitis with Kidney-yang De ficiency / 中药新药与临床药理

Objective To find a new method for the establishment of allergic rhinitis(AR) models with kidney-yang deficiency.Methods Fourty-five Hartely guinea pigs were randomized into 4 groups: hydroxycarbamide+ ovalbumin(OV); predisone+ OV;hydroxycarbamide+OV+Shenqi decoction;normal control).The general behavior and the body weight of guinea pigs were observed before and after treatment.After 3 weeks,the reaction of guinea pigs to histamine nasal drops was observed and then all animals were sacrificed to examine the pathological changes of nasal septum mucosa.Result The new model induced by hydroxycarbamide and OV was proved to be set up successfully: the manifestations of kidneyyang deficiency in the new model were more representative than those induced by predisone and OV,and the time for modeling was shorter.Conclusion Using hydr oxycarbamide and OV to set up kedney-yang deficiency AR models is an new feasible method.The result of this study will provide evidence for the further research of kidney-yang deficiency AR.

Effect of Buzhong Yiqi Decoction for Allergic Rhinitis with Spleen-qi Deficiency in Guinea Pigs / 中药新药与临床药理

Objective To investigate the effect of Buzhong Yiqi Decoction for allergic rhinitis(AR)with spleen -qi de-ficiency in guinea pigs.Methods AR guinea -pig model was established by intraperitoneal injection of ovalbumin for general sensitization and then by nasal drip of ovalbumin for local sensitization.Spleen -qi deficiency mod el was induced by gastric infusion of cold -maceration extract of Radix et Rhizoma Rhei.Fifty -four guinea pigs were randoml y allocated to four groups:AR group(Group A),AR model with spleen -qi deficiency group(Group B),AR model with spleen -qi deficiency treated by Buzhong Yiqi group(Group C)and blank control group(Group D).Nasal symptoms were observed.The number of eosinophils(EOS )in nasal secretion was detected by sm ear cytologic examination and the number of EOS and mastocyte in nasal mucosa was exa mined by pathological section.Results Before local sensitization,nasal s ymp-toms such as nose scratching,sneezing and nasal discharge in Group A,Gro up B and Group C were severer than those in Group D and were improved in Group C in the late stage.Positive rate of EOS i n nasal secretion and nasal mucosa of Group C and Group D was lower than that in Group A and Group B and the differen ce was insignificant between Group C and Group D.The number of mastocyte i n Group C was lower than that in Group A and Group B.Conclusion Buzhong Yiqi Decoction can relieve nasal symptom s in AR Guinea pigs with spleen -qi deficiency by counteracting the infiltration of EOS and the increase of nastocytes in nasal mucosa.

Comparison of pharmacological effects of tetramethylpyrazine and deuterated tetramethylpyrazines / 中国药理学通报

TMP. The results suggested that 12D-TMP and 6D-TMP possessed remarkable isotope effects, and that the pyrazine ring of TMP may be responsible for its pharmacodynamics, while the substituted groups might primarily govern its pharma-cokinetics and toxicity. The results also indicated that to study the structure activity relationship of TMP and to modify its chemical structure are very fruitful for the development of TMP analogues of high potency.