Objective To approach a way to induce MSCs to dopaminergic neuron by mesencephalic conditional media and cytokines in vitro,and supply an ideal cells source for the treatment of Parkinson's disease.Methods The rat MSCs were isolated primarily from the femurs and tibias of the Wistar rats.MSCs were cultured,proliferated and purified by passage culture.Cultuered MSCs were divided into the control and the experimental group.In control group,MSCs were cultured without any induction medium.MSCs of experimental group were first cultured at medium containing bFGF for 24 hours.Then media were replaced with induction media which contained the agents as follows,respectively:GM_1,GDNF, GDNF+GM_1,GDNF+GM_1+mesencephalic conditional media.The surface markers of the differentiated neuron,such as NSE and TH were detected by immunocytoehemistry after MSCs were cultured in induction media for 3 and 7 days.Results In control groups,the NSE expression of MSCs was very lower than experimental groups.The percentage of NSE-positive cells of GDNF+GM_1+mesencephalic conditional media group in 7 day((45.257?5.999)/HP)was significantly more than other groups(control group is 2.214?0.779,GM_1 group is 22.014?3.624,GDNF group is 31.345?2.850,GDNF?GM_1 group is 40.314?4.203,P

Parkinson's disease(PD) is a common neurodegenerative disorder with no effective protective treatment,characterized by a massive degeneration of dopaminergic neurons in the substantia nigra(SNpc) and the subsequent loss of their projecting nerve fibers in the striatum.The major neurochemical manifestation of this disorder is the loss of the neurotransmitter dopamine(DA) in the striatum as a result of the progressive degeneration of the dopaminergic neurons in the substantia nigra.There have been significant progresses in recent years reporting on the use of mesenchymal stem cells(MSCs)in gene therapy,with specific application towards PD.MSCs,a kind of multipotent adult progenitor cells,are considered as a useful vehicle for cell and gene therapy because of their multiple differentiation potentiality and self-transplantation.The present study was focused on treating rat model of PD using human tyrosine hydroxylase gene(hTH),human aromatic L-amino acid decarboxylase gene(hAADC) and human GT Pcyclohydrolase I gene(hGCH-I) engineered MSCs,in order to provide a better understanding about the application of these cells in the therapeutic benifit of PD.The gene of hTH,hAADC and hGCH-I were introduced via recombinant adeno-associated virus(rAAV) infection into the MSCs in vitro.The genetically modified MSCs expressing hTH,hAADC and hGCH-I were transplanted into the striatum of PD rat models.The behavior,the nigra-striatal level of DA and its metabolite were detected.The results of present study were shown as follows:hTH,hAADC,hGCH-I and LacZ gene were transfected into MSCs with adeno-associated virus vectors.The HEK293 packaging cells(ATCC) were transfected with the plasmids of pAAV-hTH,pAAV-hAADC,pAAV-hGCH-I,pAAV-LacZ,pAAV-RC,pHelper by using calcium phosphate precipitation.Titer was detected using HT1080 cells.Viral particles were collected and used to infect MSCs.The purified modified MSCs expressing the three kinds of genes were selected separately and were grafted in the striatum of the PD model rats in the lesion side.The MSCs genetically modified suvived well 12 weeks after transplantation.The improvements of the behavior were observed every week after transplantation.Compared with the control group,the rounds of asymmetric rotation after apomorphine administration decreased in the groups double or triple genes engineered MSCs grafted(p

Objective To investigate the clinical value of MRI in evaluating the cardiac structures and function of congenital heart diseases.Methods Fifteen cases with surgically proved different types of CHD were examined by the techniques of magnetic resonance imaging ( MRI) and echocardiography (ECG).The results were compared to determine their values in showing the abnormalities of cardiac structures and function.Results In 15 cases,20 malformations were diagnosed correctly by MRI within 24 abnormalities confirmed by operation(20/24),21 by ECG (21/24).MRI was better than ECG in displaying great vascular diseases, while ECG was better in valvular diseases.There was no significant difference between MRI and ECG in examining cardiac function.Conclusion MRI can clearly show the abnormalities of cardiac anatomy and function.

Objective To investigate the combined use of osteoarthritis treatment effect of lornoxicam and peptide drugs.Methods 260 patients with osteoarthritis treated in our hospital from May 2014 to June 2016 were selected, were randomly divided into study group and control group, each group of 130 cases.The patients in the control group were treated with sodium hyaluronate and lornoxicam, the study group was given bone peptide on the basis of the control group. The pain scores, knee function and adverse reactions were compared between the two groups.Results There was no significant difference in VAS and WOMAC scores between the two groups before treatment; after treatment, the improvement of VAS and WOMAC score in the study group was better than that in the control group (P<0.05). There was no significant difference in the incidence of adverse drug reactions between the two groups. Conclusion Sodium hyaluronate, bone peptide, lornoxicam and other drugs in the treatment of patients with osteoarthritis clinical efficacy is obvious, and fewer adverse reactions.

ObjectiveTo analyze the risk factors of nosocomial infection of patients with spinal cord injury.MethodsRetrospective surveys of nosocomial infection including risk factors, pathogen analysis and drug sensitivity tests were carried out for 88 cases with spinal cord injury in our hospital from 1996 to 2002.ResultsIt showed that the rate of nosocomial infection was 90.91% with 46.59% of urinary tract infection and 32.95% of respiratory tract infection. Nosocomial infection was closely related with such factors as the disease of spinal cord, the use of antibiotics, invasive operation, age and disturbance of disease.ConclusionReducing risk factors and using antibiotics reasonably are effective measures to reduce nosocomial infection of patients with spinal cord injury.

In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g^-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.

Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism remains unclear.To investigate the effect of synuclein on mitochondrial dysfunction induced by rotenone.The human dopaminergic SH-SY5Y cells were used as a cell model.The cells over-expressed the wild-type ?-synuclein were treated with complex I inhibitor rotenone.The cell viability,complex I activity,Mitochondrial swelling and O2-content were tested at different time point-1w,2w,4w after rotenone treated.CCK-8 test results showed that the cell viability of overexpressed ?-synuclein(SH-SY5Y-Syn)was much lower than the control group(SH-SY5Y-Ctr).After administrating with rotenone about 1w or 2w the cell viability of SH-SY5Y-Syn became higher than that of SH-SY5Y-Ctr.On the 4th week the results were contrary to the first 2 weeks.Similar results were got when test the mitochondrial function.In the first 2 weeks after roteoone administrating,the mitochondrial function of SH-SY5Y-Syn was better than that of SH-SY5Y-Ctr.This suggest that the ?-synuclein could protect the mitochondrial against the injury induced by rotenone in the early stage-1w,2w,while this effect disappeared in the final stage-4w.

Near-infrared spectroscopy ( NIR ) is widely used in the area of food quantitative and qualitative analysis.Variable selection technique is a critical step of the spectrum modeling with the development of chemometrics.In this study, a novel variable selection strategy, automatic weighting variable combination population analysis (AWVCPA), was proposed.Firstly, binary matrix sampling (BMS) strategy that gives each variable the same chance to be selected and generates different variable combinations, was used to produce a population of subsets to construct a population of sub-models.Then, the variable frequency ( Fre) and partial least squares regression ( Reg) , which were two kinds of information vector ( IVs) were weighted to obtain the value of the contribution of each spectral variables, the influence of two IVs of Rre and Reg was considered to each spectral variable.Finally, it used the exponentially decreasing function ( EDF) to remove the low contribution wavelengths so as to select the characteristic variable.In the case of near infrared spectrum of beer and corn, the prediction model based on partial least squares ( PLS ) was established.Compared with other variable selection methods, the research showed that AWVCPA was the best variable selection strategy in the same situation.It had 72.7% improvement compared AWVCPA-PLS with PLS and the predicted root mean square error (RMSEP) decreased from 0.5348 to 0.1457 on beer dataset.It had 64.7% improvement compared AWVCPA-PLS with PLS and the RMSEP decreased from 0.0702 to 0.0248 on corn dataset.

OBJECTIVETo observe the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer, and study its anti-tumor mechanism.

METHODIn vitro, MTT assay and scratch assay were adopted to detect the effect of alcohol extracts from Pharbitidis Semen on the proliferation and metastasis of Lewis lung cancer cells. The cell autophagy was detected by the acridine orange staining. The gap-junction intercellular communication (GJIC) was investigated by the fluorescent yellow transfer. The expression of aquaporin 1 (AQP1) was analyzed by the Western blotting. In vivo, the subcutaneous implant model and the experimental pulmonary metastasis model of Lewis lung cancer in mice were established to evaluate the anti-tumor and anti-metastasis effects of alcohol extract from Pharbitidis Semen. The serum carcinoembryonic antigen (CEA) and beta2 microglobulin (beta2-MG) of mice bearing Lewis lung cancer were detected by the electrochemiluminesence immunoassay. The expressions of lung AQP1 and Connexin 43 (Cx43) were examined by the immunohistochemical method.

RESULTIn vitro, alcohol extracts from Pharbitidis Semen inhibited the cell proliferation in a dose-dependent matter, significantly prevented the cell migration, down-regulated AQP1 proteins of cells, promoted GJIC, and decreased the serum-free autophagy of tumor cells. In vivo, compared with untreated model mice, alcohol extracts from Pharbitidis Semen inhibited the tumor growth in a dose-dependent matter, prevented the tumor metastasis and prolonged the life span of mice bearing Lewis lung cancer, while decreasing serum CEA and beta2-MG of mice bearing Lewis lung cancer, enhancing the immumohistochemical staining intensity of Cx43 and weakening aquaporins AQP1 positive intensity.

CONCLUSIONAlcohol extracts from Pharbitidis Semen could prevent the proliferation and metastasis in Lewis lung cancer cells. Its mechanism may be related to the promotion of GJIC and the down-regulation of AQP1.

Animals ; Antineoplastic Agents ; administration & dosage ; Aquaporin 1 ; genetics ; metabolism ; Carcinoma, Lewis Lung ; drug therapy ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Connexin 43 ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Ipomoea ; chemistry ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Metastasis ; Seeds ; chemistry

Objective To investigate the mechanisms underlying neuroprotection of silent information regulation 2 homolog 3 ( SIRT3 ) against hypoxia via preconditioning.Methods PG12 cells were randomly divided into control,hypoxic preconditioning ( Hyp),Hyp with oxygen-glucose deprivation (OGD) and OGD.MTT assay and DAPI staining were used to evaluate cellular viability.MitoSOX Red was used to measure the production of mitochondrial superoxide.The protein expression of SIRT3,PGC-1α and MnSOD were assessed by Western blot.Recombinant SIRT3 was also given to further investigate its roles in hypoxic preconditioning.Results The preconditioned PC12 cells had a higher survival rate.When expressed as a percentage of the control group,MTT values following 6 h OGD were around 51.0% in the OGD group but around 74.7% in the Hyp + OGD group ( F = 56,P ＜ 0.01).Mitochondrial ROS after Hyp was less than the OGD group.Both Hyp + OGD and OGD increased the expression of SIRT3,PGC-1α and MnSOD proteins,and these increases were greater after Hyp + OGD.Similarly,the application of recombinant SIRT3 to OGD also further increased the expression of these proteins.Conclusions Hypoxic preconditioning can protect PC12 cells against hypoxic injury.One possible mechanism of hypoxic preconditioning is via SIRT3 to upregulate PGC-la and,in turn,MnSOD to reduce generation of ROS.