Objective To investigate the effect of purified single clonally mesenchymal cells (SCMSCs) on cardiac function in rat models and also try to find out wheter SCMSCs serve as a better source for transplantation than UMSCs, BM-MNCs and PB-MNCs. Methods SCMSCs were isolated, cultured, purified, cloned and expanded. UMSCs were isolated primarily by their tight adherence to the culture dishes. BM-MNCs and PB-MNCs were prepared by Ficoll-Hypaque gradient centrifugation. All cell characters were verified by fluorescence- activated cell sorter (FACS). A total of 5?10~6 PB-MNCs, BM-MNCs, UMSCs, and SCMSCs were transplanted into the ischemic zone immediately after MI. The cardiac function was evaluated by hemodynamic technique 1 month after the transplantation. The vessel density and cell differentiation were determined by histological techniques. Results SCMSCs expressed over 99% of the mesenchymal cell surface protein and none of the hematopoietic stem cell surface protein. Hemodynamics showed that transplantation of snMSC led to the greatest improvement in cardiac function, compared with PB-MNCs, BM-MNCs, and UMSCs transplantation. In consistence with cardiac function recovery, SCMSCs transplantation resulted in the greatest angiogenesis in the ischemic wall, and the greatest number of transplanted SCMSCs expressed these cardiomyocyte proteins, or vascular endothelial cell marker, in comparison with the other heterogeneous cells. Conclusion Transplantation of single clonally purified non-hematopoietic mesenchymal stem cells from human bone marrow showed the greatest imporvement in cardiac function compared to UMSC, BM-MNC, and PB-MNC in this study.

Objective To investigate the effects of emodin on the migration and proliferation of vascular smooth muscle cell (VSMC) and the metabolism of emodin in VSMC. Methods The effects of emodin on the migration or proliferation of vascular smooth muscle cell were measured by “transwell" migration system and MTT assay. Results The migration of VSMC could be significantly inhibited by emodin and the inhibitory ratio was 83.8% in 5 ?g/mL emodin group. The antiproliferative effect of emodin was in a dose- and time- dependent manner. However, the supernatant concentration of emodin insignificantly dereased after culture for 24 h. The cytotoxicity of emodin and cellular ROS was not influenced by CYP inducer or inhibitor. The mRNA expression of emodin's primary metabolic enzyme (cytochrome p450 oxidase, CYP) up-regulated insignificantly after treatment of emodin for 24 h. Conclusion Emodin can inhibit the migration and proliferation of VSMC and can not be metabolized by VSMC. Emodin may be a choice for the medication of drug-eluting stent.

Objective To explore the function of MDA on diabetic nephropathy .Methods Glomerular mesangial cells ( GMC) were pretreated with MDA at a final concentrations of 0μmol/L, 1μmol/L, 5μmol/L, 10μmol/L and 50 μmol/L.MTT assay was used to examine the viability of GMC ) .AnnexinV-FITC was used to evaluate effect of MDA on cell apoptosis .RT-PCR and western blot were used to analyze the expression of Nrf 2, HO-1 andγGCL.Results MDA treatment inhibited GMC viability in a dose-dependent manner .MDA at the concentration of more than 5 μmol/L induced mass production of ROS in GMC ( P<0.05 ) .In addition , antioxygen of tBHQ may relieve MDA-induced reduction of cell viability .MDA inhibited the expression of HO-1 , γGCLand Nrf2 ( P <0.05 ) .Conclusions MDA inhibites GMC viability and promotes the cell apoptosis by ROS production through in-hibiting Nrf2/HO-1-γGCL.

Objective: To study the distribution,combination law of syndrome factors and characteristics of base syndromes of pneumonia based on document data.Methods: Combined methods of computer retrieval and man-made retrieval were adopted,the document data from 1981 to 2004 on lower respiratory tract infection was gathered by way of document database index.And the data was analyzed by description,Logistic regression and clustering analysis.Results: 32 documents and 141 items were used for analysis.The results indicated that among the 17 syndrome factors related to syndrome differentiation of pneumonia,the highest frequency of location syndrome factor was lung;the frequency of f ire,heat and yin-def iciency was higher;pathogenic wind,pathogenic heat and phlegm were the main etiological factors.In the document study,there were four patterns in the combinative model of syndrome factors,the combination of two syndrome factors and the combination of three syndrome factors were the main patterns,the cumulative percentage was 95.7%.Meanwhile,the main symptoms and the sub-symptoms of the mainly syndromes were screened out by adopting statistical description,logistic regression and clustering analysis.Conclusion: The main syndrome types and their inscapes of pneumonia can provide enough references for the next step research.

Objective To improve the methods for in vitro generating dendritic cells (DCs) from mouse bone marrow and to identify it with morphological, phenotype determination. Methods Cells isolated from mice bone marrow were cultured in GM-CSF (20 ng/ml), differentiating into dendritic cells. Morphological changes were observed by optical phase contrast microscopy, and surface molecules including CD11 C, CD80, MHCⅡ were detected by FACS. Results A large number of typical DCs were observed after culturing for 10 d. FACS analysis showed that the amplified DCs could express CD11 C, CD80, MHCⅡ. Conclusion A large quantity of highly pure BM-DC can be obtained by this method.

This paper has analysed the particularity of practice and teaching of oral and maxillofacial surgery.Binding with the change of medical pattern in recent years,the change of relation of doctors and patients as well as the present of "the People's Republic of China Practical Doctor Law"and "Rules of Sitling Malpractice",it has discussed that we should simultaneously think highly of raising to practice ability, reinforcing education of medical ethics, emphasizing legal consciousness and deepening ethics idea during practice teaching of oral and maxillofacial surgery,which can ensure well development of interns.

Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.

Objective To observe the effect of acupuncture combined with integrated Chinese-Western neuromuscular facilitation technique on persistent vegetative state (PVS).Methods41 PVS cases were randomly divided into the observation group (n=21) and control group (n=20). The observation group was treated with acupuncture, neuromuscular facilitation technique combined with reasonable obligatory exercise, neuromuscular electrical stimulation, manipulation treatment and Chinese medicine. The control group was treated with the Western and Chinese medicines, and hyperbaric oxygen.ResultsAfter 1～3 months treatment, 7 cases cured,8 cases were markedly effective, 5 cases were effective, 1 cases were ineffective in the observation group with a total markedly effective rate (71.4%) and effective rate (95.2%). While, in the control group, 3 cases cured, 5 cases were markedly effective, 6 cases were effective, 6 cases were ineffective and total markedly effective rate was 40%, effective rate was 70%. There was a significant difference between the two groups in the total markedly effective rate and the effective rate ( P<0.05). The average PVS score increased by 7.46±1.22 in the observation group and 4.59±1.21 in the control group. Also there was a significant difference between the two groups ( P<0.001).ConclusionThe therapy of acupuncture combined with integrated Chinese-Western neuromuscular facilitation technique can markedly promote PVS patients coming round and improve patients' prognosis.

[ ABSTRACT] AIM:To investigate the effect of artemisinin on lipopolysaccharide ( LPS)-induced intestinal epi-thelial barrier damage in IEC-6 cells and its molecular mechanism.METHODS:Cultured IEC-6 cells were divided to 5 groups:control group, LPS (100 mg/L) group and LPS +Artemisinin (30, 50 and 100μmol/L) groups.The cytotoxici-ty was detected by MTT assay.The releases of TNF-α, IL-1βand IL-6 in the IEC-6 cells were measured by ELISA.The transepithelial electrical resistance ( TER) was detected by electrical resistance tester, and the horseradish peroxidase (HRP) flux permeability were analyzed by a microplate reader.The expression of tight junction proteins, ZO-1, claudin-1 and occludin, and the expression of TLR4/MyD88/NF-κB at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:Artemisinin alone (up to 100 μmol/L) or in combination with LPS (100 mg/L) was not toxic to IEC-6 cells.Compared with control group, the releases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells significantly increased after treatment with LPS.The expression of TLR4/MyD88/NF-κB was activated by LPS.LPS down-regulated the protein expression of ZO-1, claudin-1 and occludin.However, artemisinin treatment decreased the re-leases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells.The expression of TLR4/MyD88/NF-κB at mR-NA and protein levels was gradually reduced after treatment with artemisinin.In addition, artemisinin upregulated the pro-tein expression of ZO-1, claudin-1 and occludin significantly (P<0.01) in a dose-dependent manner.CONCLUSION:Artemisinin attenuates LPS-induced intestinal epithelial barrier damage by inhibiting TLR4/MyD88/NF-κB activation in the IEC-6 cells.

AIM:To study the effects of extracellular potassium on the protein expression of wild-type HERG and its mutant L539fs/47.METHODS:Wild-type HERG (WT) or its mutant HERG-L539fs/47 (MT) were transfected into HEK293 cells for 36 h.The cells were incubated in different media containing 0.8, 4.3 or 10 mmol/L potassium.Af-ter 6 h of incubation, the protein expression of HERG was detected by flow cytometry.After 12 h of incubation, the locali-zation and quantity of the proteins were detected by laser confocal imaging and Western blot.RESULTS: Different from the retention of mutant protein in cytoplasm, wild-type HERG protein was mainly distributed in the cell membrane.The 2 proteins both increased with the changes of extracellular potassium.Flow cytometry showed that the fluorescence in the 2 groups both increased with the changes of extracellular potassium ( P<0.01 ) .The fluorescence in WT group was signifi-cantly higher than that in MT group (P<0.01).Western blot showed that mutant HERG protein included only one 60 kD band, different from the 135 kD and 155 kD bands in wild-type HERG, which were affected by the changes of extracellular potassium (P<0.05).CONCLUSION:The retention of HERG mutant L539fs/47 protein in the cytoplasm is more than wild-type HERG.Chronic high extracellular potassium keeps the stability of wild-type and mutant HERG proteins on the cell membrane.Chronic low potassium reduces the expression of HERG channel proteins in a time-dependent manner.