ObjectiveTo observe the effect of exercise on expression of protein kinase B (PKB) in adipose tissue of insulin resistant rats fed by high fat diet.Methods30 male Wistar rats were randomly divided into the control group (n=10),given basic diet; the model group (n=20), given fat diet. After 4 weeks, the model group was randomly divided into 2 subgroups, the insulin resistant group was continually given high fat diet, the exercise treated group accepted high fat food and swimming training. After 6 week intervention, the expression of PKB stimulated by insulin in adipose tissue was determined with Western blotting at the end of experiment.ResultsAfter long term high fat diet, expression of PKB in adipose tissue of the insulin resistant group decreased by 23.5% comparing with the control group (P<0.01). After 6 weeks swimming training, the expression of PKB of the exercise treated group was increased by 19.2% comparing with the insulin resistant group (P<0.01).ConclusionExercise treatment can significantly elevate the expression of PKB and ameliorate the state of insulin resistance.

OBJECTIVETo express and purify the protein coded by the TRAF-type zinc finger domain of myasthenia gravis (MG)-related gene P9 (P9-ZFD) and to prepare P9-ZFD antiserum for detecting expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of patient with MG.

METHODSThe cDNA encoding P9-ZFD was amplified by RT-PCR. The cloned P9-ZFD cDNA was ligated into pET24a, and the P9-ZFD recombinant protein was induced via E. coli. BL21 (DE3) and purified by histidine affinity chromatography. P9-ZFD antiserum was prepared and its titer and specificity were determined by ELISA and Western blot. Expression and subcellular distribution of P9-ZFD protein in the skeletal muscles of MG and control were studied.

RESULTSThe molecular weight of purified P9-ZFD protein was about 30 kD. Its purity was more than 95%. Antiserum specific for P9-ZFD was excellent. P9-ZFD protein is fully confined to the cytoplasm membrane of skeletal muscle cell of MG, obvious immunostaining was absent in the A, I, and Z bands of cytoplasm and no immunoreactivity was observed in the skeletal muscle cell of control.

CONCLUSIONP9-ZFD protein is expressed as a cytoplasm membrane-bound protein and has obvious distribution difference in the skeletal muscle cells of patient with MG and normal control.

Adult ; Cell Membrane ; metabolism ; Escherichia coli ; metabolism ; Female ; Humans ; Muscle Proteins ; biosynthesis ; genetics ; Muscle, Skeletal ; metabolism ; pathology ; Myasthenia Gravis ; metabolism ; Peptide Fragments ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; Zinc Fingers

Objective To develop a pharmacy device for drug counting and dispensing.Methods The device was composed of four parts of dispensing tank, hook, support frame and separator, which was made of bamboo and metal. The multi application of the separators contributed to fulfilling the functions of the device.Results The device gained advantages over the traditional mode in time,accuracy,drug loss and the staff's efficiency.Conclusion The device is easy to manufacture and operate and behaves well in feasibility, practicability and innovation, and thus is worthy promoting practically. [Chinese Medical Equipment Journal,2018,39(5):47-49]

OBJECTIVETo explore the role of reversal multidrug resistance (MDR) using short hairpin RNA (shRNA) expression vectors in multidrug resistance human leukemia cell line K562/ADM.

METHODSThe oligonucleotides with 19-mer hairpin structure were synthesized. The shRNA expression vectors were constructed and introduced into K562/ADM cells. Expression of mdr1 mRNA was assessed by RT-PCR, and P-gp expression was determined by Western blot. The apoptosis and sensitivity of the K562/ADM cells to doxorubicin were quantified by flow cytometry and methyl thiazolyl tetrazolium (MTT) assays, respectively. Cellular daunorubicin accumulation was assayed by laser confocal scanning microscope (LCSM).

RESULTSIn positive clones of K562/ADM cells stably transfected with pSilencer 3.1-HI neo mdr1-A and mdr1-B shRNA expression vectors, RT-PCR showed that mdr1 mRNA expression was significantly reduced to 35.9% (P < 0.05), 27.5% (P < 0.01), respectively. Western blot showed that P-gp expression was significantly and specifically inhibited. Resistance against doxorubicin was decreased from 79-fold to 38-fold (P < 0.05), 30-fold (P < 0.01) respectively. Furthermore, the fluorescence intensity of K562/ADM cells was increased significantly compared with the control. shRNA vectors significantly enhanced the cellular daunorubicin accumulation. The percent of the apoptosis cell was significantly enhanced to 18.1% (P < 0.05) , 54.4% (P < 0.01) respectively.

CONCLUSIONSshRNA expression vectors can effectively reverse MDR, and restore the sensitivity of drug-resistance K562/ADM cells to conventional chemotherapeutic agents.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

Objective To assess the prevalence rates of intra-and extraeranial large-artery stenosis in the rural community population and its related risk to the development of stroke.Methods The study subjects included 1337 residents in the rural community of Beijing.Transcranial Doppler Was carried our to examine the relation between intra-and extracranial large-artery stenosis and subsequent cerebrovascular events, with a meall follow-up period of 16.7 months.Results The incidence densities of cerebral infarction and cerebral hemorrhage in persons without cerebral large-artery stenosis were 410.6 and 351.9/100-thousand person-years,respectively.In the group wlth large-artery stenosis,the incidence density of cerebral infarction was 3303.7/100-thousand person-years.Data from The Fisher's Exact test showed a significant difference in the two groups (P=0.004).Cerebral large-artery stenosis(OR=6.593,95%CI:1.712-25.390)and smoking (OR=8.437,95% CI:2.327-30.598)appeared to be independent risk factors to cerebral infarction.Conclusion Cerebral large-artery stenosis and smoking were independent risk factors to cerebral infarction.

OBJECTIVETo observe the clinical effect of Shengxueling (SXL) on idiopathic thrombocytopenic purpura (ITP), and study the possible mechanism.

METHODSEighty-six cases of ITP were randomly divided into two groups. The SXL group, 56 patients treated with SXL, a traditinal Chinese medicine and 30 patients administered with prednisone were taken as control. Each group took drugs for 3 months and was under follow-up observation.

RESULTSIn the SXL group, the total effective rate was 85.71%, similar to prednisone 83.33% (P > 0.05) for 3 months, but the total effective rate of SXL (91.07%) were obviously better than that of the control group (53.33%) (P < 0.01) for 6 months and had no obvious adverse reaction. The patients bleeding was alleviated or stopped, the general condition was improved. At the same time, blood platelet count (PLT) was increased, platelet associated immunoglobulin (PAIg) and interleukin-4 (IL-4) were markedly dropped, the level of natural killers cells activity (NKa) increased, the rate of T lymphocyte subsets gradually returned to normal level. Megakaryocyte tended to maturation on bone marrow smear after treatment. All differences above were statistically significant.

CONCLUSIONSXL is an effective and safe medicine for ITP. Its mechanism could regulate cytoimmune, inhibit platelet antibody to reduce the destruction of platelet, increase the number of platelet, promote the division and maturation of megakaryocyte, facilitate the production and release of platelet, lower the fragility of capillary, prevent and cure hemorrhagic tendency.

Adolescent ; Adult ; Blood Platelets ; metabolism ; Bone Marrow ; pathology ; Female ; Glucocorticoids ; therapeutic use ; Hemorrhage ; etiology ; physiopathology ; Humans ; Immunoglobulins ; blood ; Interleukin-4 ; blood ; Killer Cells, Natural ; pathology ; Lymphocyte Count ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Phytotherapy ; adverse effects ; Plant Preparations ; adverse effects ; therapeutic use ; Platelet Count ; Prednisone ; therapeutic use ; Purpura, Thrombocytopenic, Idiopathic ; blood ; complications ; pathology ; therapy ; T-Lymphocyte Subsets ; pathology ; Treatment Outcome

AIM:To investigate the effect of bone marrow mesenchymal stem cell(BMSC) transplantation on learning and memory abilities and pathological changes of Alzheimer disease (AD) mice and the molecular mechanisms. METHODS:C57/BL6 wild-type (WT) and transgenic(Tg) mice were randomly divided into 4 groups:WT/PBS group, WT/BMSCs group,Tg/PBS group and Tg/BMSCs group. The mice were administered with PBS or BMSCs via intracere-broventricular injection. Spatial learning and memory abilities of the mice were evaluated by Morris water maze test on the 3rd day after surgery. Real-time PCR was applied to detect the mRNA expression of CX3C chemokine ligand 1 (CX3CL1),CX3C chemokine receptor 1 (CX3CR1), IL-1β, TNF-α, Nurr1, YM1, insulin-degrading enzyme (IDE) and matrix metalloproteinase 9(MMP9). The protein levels of CX3CL1 and Aβ42 were measured by ELISA. Western blot was used to detect the protein expression of postsynaptic density protein 95 (PSD95) and synaptophysin (SYP). RE-SULTS:The transplanted BMSCs were observed near the hippocampus of APP/PS1 mice on the 10th postoperative day. The escape latency of the mice in Tg/PBS group was significantly longer than that in the WT/PBS mice(P<0.05). Com-pared with Tg/PBS group,the escape latency of Tg/BMSCs group was significantly shorter (P<0.05), and the mRNA and protein levels of CX3CL1 in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.01). The results of immunohistofluorescence staining showed that BMSC transplantation promoted the activation of microglia in the brain of WT and Tg mice. The mRNA expression of YM1 was up-regulated in WT/BMSCs group and Tg/BMSCs group (P<0.05). Compared with WT/PBS mice, the mRNA expression of TNF-α in the cortex and hippocampus of Tg/PBS group was significantly increased (P<0.05),and the mRNA expression of Nurr1 in the cortex was significantly decreased (P<0.01). Meanwhile,the mRNA expression of TNF-α in the cortex of Tg/BMSCs mice was decreased(P<0.01) and the mRNA expression of CX3CR1 and Nurr1 was up-regulated compared with Tg/PBS group (P<0.05). The results of Western blot showed that the protein levels of PSD95,p85,p110 and p-Akt in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.05). Finally, BMSC transplantation reduced the protein level of Aβ42 in APP/PS1 mice(P<0.05), and increased the mRNA expression of IDE and MMP9 in the hippocampus (P <0.05). CONCLU-SION:BMSC transplantation modulates neuroinflammatory responses and promotes neuroprotective factor and synaptic pro-tein expression,thus improving the learning and memory abilities in the APP/PS1 mice,which may be achieved by up-reg-ulating the expression of CX3CL1.

OBJECTIVETo study the mechanism of acupuncture and moxibustion in adjusting apoptosis of colonic epithelial cells of the rat of ulcerative colitis (UC).

METHODSUC rats were randomly divided into four groups including normal control group (NC), model control group (MC), herbs-partitioned moxibustion group (HPM) and electroacupuncture group (EA). The HPM and EA groups were respectively treated with herbs-partitioned moxibustion and electroacupuncture at "Qihai" (CV 6) and bilateral "Tianshu" (ST 25). Then the colon of the rat was taken and the changes of the tissue structure and the cell apoptosis were observed by electron microscope and flow cytometer respectively.

RESULTSThe colon tissue showed histopathological changes, and apoptosis of epithelial cells increased in the model mice of UC as compared with the normal rat. Electroacupuncture and drug-separated moxibustion could significantly decrease the abnormal apoptosis of epithelial cells.

CONCLUSIONThe adjustment of the abnormal apoptosis of epithelial cells is one of the mechanisms of drug-separated moxibustion and electroacupuncture treating UC.

Acupuncture Points ; Animals ; Apoptosis ; Colitis, Ulcerative ; therapy ; Epithelial Cells ; Humans ; Moxibustion ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the prognostic significance of a new grading and scoring system (based on the new IASLC/ATS/ERS classification) in stage I pulmonary adenocarcinoma, as compared with the WHO grading system.

METHODSThe clinicopathologic characteristics of 125 patients with stage I pulmonary adenocarcinoma primarily treated by surgical resection were reviewed retrospectively. All cases were classified according to the new IASLC/ATS/ERS classification and graded into three prognostic groups based on the new classification, the Sica scoring system and the WHO grading system, respectively. The differences in prognosis of the three groups were analyzed.

RESULTSThere was a statistically significant correlation between the new grading system and the WHO grading system (P = 0.000). Both of them showed negative correlation with overall survival. The new scoring system however better correlated with disease recurrence and/or metastasis (P = 0.855, P = 0.073 versus P = 0.011). According to univariate Log-rank test, the prognosis correlated with tumor size (P = 0.004), clinical stage (P = 0.000), the WHO grading (P = 0.020), the new grading system (P = 0.000), the new scoring system (P = 0.000), vascular invasion (P = 0.021), and recurrence and/or metastasis (P = 0.000). The Cox regression analysis demonstrated that clinical stage (P = 0.014), the new grading system (P = 0.047), the new scoring system (P = 0.043), and recurrence and/or metastasis (P = 0.018) were significantly independent poor prognostic factors.

CONCLUSIONSThe new grading and scoring system shows good correlation with the WHO grading system. Compared with the WHO grading system, the new scoring system based on the new IASLC/ATS/ERS classification provides valuable information in categorizing stage I pulmonary adenocarcinoma cases with different risks of disease recurrence, tumor metastasis and prognosis.

Adenocarcinoma ; classification ; pathology ; surgery ; Adenocarcinoma, Mucinous ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma in Situ ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; classification ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Grading ; methods ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Research Design ; Retrospective Studies ; Societies, Medical ; Young Adult

Animals ; Collagen Type I ; metabolism ; Collagen Type II ; metabolism ; Collagen Type III ; metabolism ; Female ; Kinetin ; pharmacology ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism