This article systematically introduced the development of Smart Healthcare mode in Hangzhou. From the aspects of connotation, pattern, characteristics and the change of medical treatment mode, the authors pointed out that the development of Smart Healthcare must follow such principles as satisfaction of the people, enhanced inter-departmental synergy, government-guided social participation, and full involvement of medical workers.

CDKs proteins are a kind of cell cycle protein-dependent kinases, which serve as important roles in controlling cell division and transcriptional stages. Among them, CDK9, as a key regulator responsible for the transcriptional elongation of cells, drives the development of various malignant cells and is considered as an important target in the field of anti-tumor drug development. However, the CDK family proteins feature high conservativeness and similarity in structure, leading to the poor selectivity and severe side effects for traditional small-molecular CDK9 inhibitors, which has limited their clinical applications. In view of this, there is an urgent need to investigate CDK9 targets through a novel strategy. The PROTAC is an emerging drug discovery strategy that the degrader could specifically recognize the target protein through indirect linkage with ubiquitin ligases and ultimately eliminate the target protein through the ubiquitination degradation system. This paper provides a brief overview of the structure and function of CDK9 protein, its relationship with the poor prognosis of clinical diseases, as well as the currently reported small molecular inhibitors. The latest research progress on the targeted degradation of CDK9 protein based on PROTAC technology is highlighted. Finally, the development prospects of this target protein in this novel technology field are summarized and prospected, aiming to provide a reference for the development of antitumor drugs in this direction.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

Objective To evaluate the relationship between vascular endothelial function of coronary heart disease and the severity of coronary artery disease(CHD).Methods 73 patients undergoing coronary angiography were divided into two groups:CHD group(n=39)and non-CHD group(n=34)according to the result of coronary angiography.13 healthy subjects without risk factor of CHD were chosen as normal control group.FMD and NTG-MD of endothelial-dependent and independent dilatation of brachial artery were measured by using of high frequency linear-array uhrasonography for assessment of the vascular endothelial function.and the relationship between the vascular endothelial function and the severity of coronary artery stenosis was analyzed.Results A significant difference was obtained in the FMD among the CHD group,non-CHD group and control group[(4.81±2.33)%vs.(9.29±3.88)%vs.(13.58±1.80)%,F=48.012,P＜0.01).Significant difference was shown in NTG.MD among the three groups[(13.72±3.27)%vs.(15.64±2.65)%vs.(16.54±2.98)%,F=6.015,P＜0.01]and significant difference was shown between CHD group and the other two group(P＜0.05).The FMD was negatively correlated with the basdine value of the brachial artery.tlle number of stenotic coronary ateries and the severity of coronary artery stenosis(r=-0.224,-0,316,-0.721,P=0.038,0.003 and ＜0.001).NTG-MD was also negatively correlated with the baseline value of the brachial artery,the number of stenotic coronary arteries and the severity of coronary artery stenosis(r=-0.483,-0.258,-0.372,P＜0.001,0.027,0.001).Stepwise regression analysis displayed a linear relationship between FMD and the severity of coronary artery stenosis,the baseline value of the brachial artery(r=-0.012,-0.022,P＜0.001).NTG-MD was linearly related to the baseline value ofthe brachial artery and the severity of coronary artery stenosis(r=-0.032,-0.0073,P＜0.001).Conclusion The degree of damage of endothelial function of coronary heart disease is linearly correlated with severity of coronary arteIT stenosis.

Objective: To construct and identify a lentiviral vector harboring RNAi sequence targeting the human high mobility group A1(HMGA1) gene.Methods: The effective sequence of siRNA targeting the HMGA1 gene confirmed in our previous study,the complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCL-GFP vector diced by the restriction enzyme of HpaⅠ and XhoⅠ,which contained the U6 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing HMGA1 shRNA was named LV-sh HMGA1 and confirmed by PCR and DNA sequencing.A total of 293T cells were cotransfected with LV-sh HMGA1,pHelper 1.0 and pHelper 2.0.All the virus stocks were produced by Lipofectamine2000-mediated transfection.The titer of the virus was tested according to the expression level of GFP.Results: PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the human HMGA1 gene was successfully inserted into the lentiviral vector.The titer of the recombinant lentiviral vector was 5?107 TU/ml.Conclusion: The successful construction of the lentiviral vector of HMGA1 has prepared the ground for further studies on the functions of the HMGA1 gene with the RNAi technique.

Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.

BACKGROUND: Two-dimensional electrophoresis (2-DE) is precise and effective for the isolation and analysis of complex protein mixtures, has become one of valuable key techniques for proteome. Mice models were as research subjects to model human disease status, one of important methods for cardiovascular disease. The establishment and optimization of 2-DE for proteomes of mice myocardium will provide basis for heart disease.OBJECTIVE: To establish and optimize a stable technique of 2-DE for proteomic analysis of mice myocardium.DESIGN: Mice were used as research subjects, observational comparative study.SETTING: Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University.PARTICIPANTS: The experiment was performed at the Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University between February and September 2004. Totally 12 male BABL/c mice of 4-5 weeks without special-pathogen free with the body mass of 12-15 g were selected, which was provided by the Experimental Animal Center of Southern Medical University.METHODS: The mice were put to death by dislocation of cervical vertebra under drugged state to obtain heart. Various protein extraction method,loading sampling buffer, strip range of IPG, sampling volume of protein,staining method and so on were compared, analyzed, scanned and digilized,and then performing image analysis. Each link in the study was established and optimized.MAIN OUTCOME MEASURES: Resolution of protein and reproducibility of the trial.RESULTS: A method that was optimal for cardiac tissue can isolate about a total of 910 protein spots on pH 4-7 gel strips. 200 μg was fit for silver stain while 1000 μg was for Coomassie stains. It was found that in the myocardium of normal mice about (920±30) protein spots were obtained in 17 em pH 3-10 L gel strip, while around (880±30) were gained in the pH 4-7 L gel strip. The mean matching rate achieves to 86. 9%.CONCLUSION: The findings of the trial showed that 2-DE technique can be effectively applied for proteomics of myocardium of mice to regulate and optimize, and relatively stable separation method of myocardial protein was established.

AIM: To determine the contents of ephedrine、laetrile、glycyrrhizic acid and liquiritin in Huagai Powder traditional decoction and granule decoction(Herba Ephedrae,Semen Armeniacae Amarum,Radix et Rhizoma Glycyrrhizae,etc.) by HPLC. METHODS: An Agilent SB-C_(18) column was used for ephedrine,the mobile phase was acetonitrile-0.1% phosphoric acid(4∶96),detection wavelength at 207 nm;An Agilent XDB-C_(18) column was used for laetrile,the mobile phase was methanol-water(23∶77),detection wavelength at 215 nm;An Agilent XDB-C_(18) column was used for glycyrrhizic acid,the mobile phase was methanol-0.2 mol/L ammonium-acid(67∶33∶1),detection wavelength at 250 nm; An Agilent XDB-C_(18) column was used for liquiritin,the mobile phase was acetonitrile-0.5% acetic acid(20∶80),detection wavelength at 276 nm. RESULTS: There were discrepancies between traditional decoction and granules in the contents of four ingredients,the average contents in granules were more than those in the traditional decoction. CONCLUSION: The peaks of the four ingredients are(segregated)(effectively),and the method is simple,conveient,exact and has good reproducibility,the other ingredients do not have interferences for the determination.

Objective To investigate the value of remifentanil IV PCA joint doula in labor analgesia.Methods 480 single fetus at term vaginal delivery of maternal patients were randomly divided into four groups:120 cases voluntary implementation of remifentanil IV PCA joint doula maternity paternity were selected as the experimental group; 120 cases in control group an order to carry out the studies in our hospital childbirth select one to one Doula paternity maternal; 120 cases voluntary implementation of the control group 2 intravenous analgesia with remifentanil maternal;120 cases in control group three normal deliveries in our hospital does not take any measures of maternal.The analgesic effect,labor time,postpartum hemorrhage,cesarean section rate,neonatal asphyxia,the medical staff satisfaction were compared in four groups.Results Analgesic effect of experimental group and control group 2 was significantly better than the control group 1 and the control group 3,the differences were statistically significant (t =4.34,4.76,4.94,5.32,6.32 ;4.42,4.71,4.86,5.28,6.26,all P ＜ 0.05).The first,second,third labor stage of experimental group were (516 + 123)min,(29 + 10)min,(8 + 4)min,which were shorter than those of the other groups (t =3.76,4.21,4.18,all P ＜ 0.05).Conclusion Remifentanil IV PCA joint Doula is safe and easy for labor analgesia.

Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.