OBJECTIVESTo investigate whether antisense human telomerase reverse transcriptase (hTERT) could inhibit the activity of telomerase and the proliferation of K562 cells.

METHODSThe antisense plasmid was constructed by reverse insertion of hTERT PCR product into plasmid pLNCX-neo. Then the constructed plasmid was introduced into K562 cells by liposomes-mediated DNA transfection. The inhibition effects of telomerase on the proliferation of K562 cells were analyzed by MTT and colony formation assay, the telomerase activity of K562 cells by TRAP-PCR ELISA methods.

RESULTSThe growth rate of antisense hTERT transfected K562 cells was significantly lower than those of the controls, and the colony formation capacity of the transfected cells decreased significantly (P < 0.01), the colony number is (100.33 +/- 7.57)/10(3) cells, (92.67 +/- 5.86)/10(3) cells and (50.33 +/- 6.11)/10(3) cells for control K562 cells, K562 neo cells and antisense hTERT transfected HL60 cells, respectively. The telomerase activity of antisense hTERT transfected K562 cells was significantly inhibited.

CONCLUSIONThe expression of an antisense sequence to the mRNA sequence of telomerase protein subunit can inhibit the activity of telomerase, slow the cell growth and inhibit the capacity of colony formation of K562 cells.

Cell Division ; drug effects ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA, Antisense ; genetics ; pharmacology ; RNA, Messenger ; genetics ; Telomerase ; drug effects ; genetics ; metabolism ; Transfection

OBJECTIVETo discuss the mechanism of scar hypertrophy in adenosine receptor A(2A) (A(2A) R) knockout mice.

METHODSAnimal models of hypertrophic scar were established in 12 A(2A) R knockout mice and 12 wild-type mice as control. The thickness and the size of transverse section of the hypertrophic scar were observed by H-E staining. The hydroxyproline (HYP) in the scar was measured colorimetrically. The TGF-beta expression was tested by Western blotting method.

RESULTSThe hypertrophic scar in wild-type mice was more severe than that in knockout mice. Compared with self-control, the increase of the thickness and the size of transverse section of hypertrophic scar was markedly higher in wild-type group than in the knockout group (P < 0.01). There was significant difference in HYP content between the two groups (P < 0.01). Compared with self-control, the increase of TGF-beta expression in wild-type group was much more than that in knockout group (P < 0.01).

CONCLUSIONSThe TGF-beta expression decreases in the A(2A) R knockout mice. The scar hypertrophy is also much less in the A(2A) R knockout mice.

Animals ; Cicatrix ; metabolism ; pathology ; Disease Models, Animal ; Mice ; Mice, Knockout ; Receptor, Adenosine A2A ; genetics ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVETo observe the anti-tumor effect of silencing the expression of HIF-1alpha on cervical cancer in nude mice and to explore its mechanism of action.

METHODSHuman cervical cancer cell line Siha cells were divided into 3 groups: mock control group, control group transfected with scrambled sequence plasmid, and experimental group transfected with pU-HIF-1alpha-shRNA eukaryotic expression plasmid. Cultured cells of the three groups were inoculated in nude mice to establish cervical cancer-bearing nude mice. HIF-1alpha RNAi assay was performed to evaluate the tumor-suppressive effect of HIF-1alpha silencing on cervical cancer-bearing nude mice. Immunohistochemistry and Western blot were used to observe the distribution and protein expression of HIF-1alpha and GLUT1, while RT-PCR was adopted to detect the gene expression of HIF-1alpha, GLUT1 and HKII. The product of glycolysis (lactic acid) and apoptosis in tumor cells were detected by colorimetry and semi-quantitative TUNEL staining, respectively.

RESULTSThe tumor growth in experimental group was significantly slower than that in the two control groups (P < 0.05). On the 50th day after transplantation, the tumor weight in the experimental group was (1.90 +/- 0.28) g, significantly lower than (2.95 +/- 0.77) g in the control group and (2.54 +/- 0.56) g in the mock group (P < 0.01). In the experimental group, the gene and protein levels of HIF-1alpha were 0.45 +/- 0.04 and 1.25 +/- 0.92, and the levels of GLUT1 were 0.32 +/- 0.02 and 1.25 +/- 0.48, respectively. Both indicators in HIF-1alpha and GLUT1 were lower than that in the two control groups (P < 0.05). The expression levels of HKII gene and lactic acid in the experimental group were lower than that in the two control groups (P < 0.05), but the apoptotic cells were much more numerous in the experimental group than that in matched control groups (P < 0.01).

CONCLUSIONThe gene therapy by siRNA targeted silencing of HIF-1alpha may down-regulate its downstream genes GLUT1 and HKII expression, therefore, to reduce the tumor glycolysis activity and promote tumor cell apoptosis, and exert a tumor-suppressing effect in vivo.

Animals ; Apoptosis ; Cell Line, Tumor ; Female ; Gene Silencing ; Genetic Therapy ; Glucose Transporter Type 1 ; genetics ; metabolism ; Hexokinase ; genetics ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Plasmids ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Random Allocation ; Transfection ; Tumor Burden ; Uterine Cervical Neoplasms ; metabolism ; pathology ; therapy

OBJECTIVEThe structure-activity relationship between traditional Chinese medicine (TCM) compounds and antibacterial activity was studied by chemoinformatics approach.

METHODCytoscape and its plug-in ChemViz were applied to compute the 2D chemical structure similarity and topological parameter TPSA (topological molecular polar surface area), which measures cell permeability of chemicals, between TCM compounds and clinical antibacterials. The overall degree of structure similarity was then calculated and represented by E-value for the eight categories of TCM compounds and the known antibacterials.

RESULTOur results indicated that flavonoids showed good structural similarity with antibacterials and appropriate cell permeability, compared with those of the TCM compounds of the other categories. As flavonoids were featured by good drug safety, it suggested that they can be regarded as the preferred lead compounds skeleton structure source for further antibacterials synthesis.

CONCLUSIONThe application of chemoinformatics helps explore the structure-activity relationship between TCM compounds and the antibacterial activity and search for suitable antibacterial lead compounds skeleton structure source.

Anti-Bacterial Agents ; chemistry ; pharmacology ; Informatics ; methods ; Medicine, Chinese Traditional ; Software ; Statistics as Topic ; Structure-Activity Relationship

This study is to investigate the protective effect of quercetin against adriamycin-induced cardiotoxicity and its mechanism. The cardiotoxicity was induced by intraperitoneal injection of adriamycin (ADR) at a single dose of 20 mg x kg(-1). Mice were randomly divided into 5 groups (n=20): normal control group, ADR 20 mg x kg(-1) group, quercetin (50, 100, and 200 mg x kg(-1) groups, intragastric administration, once a day, for 7 days before ADR administration). The health conditions, electrocardiogram, activity of iNOS, SOD and LDH, levels of NO and MDA in serum or tissue homogenate, the ultrastructure and the expression of p53 protein in cardiac tissue of mice were observed. Compared with the normal control group, ADR decreased the amplitude of ECG's R wave (P < 0.001), increased the incidence of arrhythmia (to 60%), injured myocardial ultrastructure, increased the activity of LDH and iNOS, and levels of NO and MDA, decreased the activity of SOD, and increased the expression of p53 (P < 0.001). Compared with ADR 20 mg x kg(-1) group, the quercetin decreased the levels of LDH, iNOS, NO and MDA, increased the activity of SOD, restored the amplitude of R wave, decreased the incidence of arrhythmia and p53 expression (P < 0.001 , P < 0.01 or P < 0.05), and markedly reduced the myocardial ultrastructure injury. Quercetin had protective effect against adriamycin-induced cardiotoxicity. The mechanism may be related to its enhancing myocardial SOD activity, decreasing iNOS activity and inhibiting myocardial apoptosis.
Animals ; Apoptosis ; drug effects ; Arrhythmias, Cardiac ; blood ; chemically induced ; metabolism ; pathology ; Doxorubicin ; Female ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Myocardium ; metabolism ; ultrastructure ; Myocytes, Cardiac ; metabolism ; ultrastructure ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type II ; blood ; Protective Agents ; pharmacology ; Quercetin ; pharmacology ; Random Allocation ; Superoxide Dismutase ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective To investigate the clinical effect of simultaneous integrated boost intensity-modulated radiation therapy (SIB-IMRT)and whole brain radiation therapy (WBRT) plus sequential boost conformal radiation therapy (SBCRT) in the treatment of multiple metastasis tumor of brain.Methods A total of 98 patients with multiple metastasis tumor of brain in the Radiation Oncology Center of the First Affiliated Hospital of Xinxiang Medical University from August 2014 to July 2015 were divided into observation group (n =60) and control group (n =38) according to the treatment plan.The patients in the observation group were treated with SIB-IMRT,the whole brain planned target dose was 2 Gy every time,and the target dose of the metastatic target volume was 3 Gy every time for 20 times (5 times weekly).The patients in the control group received WBRT plus SBCRT,the WBRT dose was 3 Gy every time for 10 times(5 times weekly),then the metastatic tumor target area was treated with SBCRT,the prescribed dose was 3 Gy every time for 10 times.All patients were followed up from the end of treatment to December 2016.The effective rate,disease control rate and one-year survival rate were compared between the two groups.Results The patients in the two groups were successfully treated with radiotherapy.Ninety patients were followed up,eight patients were lost to follow-up,the follow-up rate was 91.8％ (90/98).The effective rate,disease control rate and oneyear survival rate in the observation group were significantly higher than those in the control group (x2 =5.371,4.352,6.002;P ＜ 0.05).The median progression free survival time in the observation group was significantly longer than that in the control group (x2 =6.537,P ＜ 0.05).There were no significant differences in the incidence of bone marrow suppression,digestive system reaction and nervous system damage between the two groups (x2 =1.821,2.032,3.782;P ＞ 0.05).Conclusion SIB-IMRT can improve the effective rate,disease control rate and one-year survival rate of patients with multiple metastasis tumor of brain.

OBJECTIVE To observe the clinical effect of Chinese medicine wet compress in the intervention of diabetic foot ulcers of meridians and collaterals heat-stasis accumulation type after interventional vascular surgery.METHODS 5 7 patients with diabetic foot ulcers were randomly divided into the treatment group(29 cases)and the control group(28 cases).Both groups were given arterial interventional therapy of lower extremity and the basic treatment.On the basis of the control group, the treatment group was also given Chinese medicine wet compress treatment.The clinical effect was observed after 8 weeks of treatment.RESULTS The total effective rate of the treatment group and the control group was 93.10% and 78.57% respec-tively,the treatment group was better than the control group(P<0.05).After the interventional vascular surgery,the ankle-brachial indexes(ABI)in both groups were significantly decreased than those of the pre-treatment(P<0.01),and P>0.05 compared between groups.TCM syndrome scores in both groups were improved after the treatment(P<0.01),and the treat-ment group was improved more significant than the control group(P<0.01).The wound area and wound volume of the two groups were significantly decreased than those of the pre-treatment(P<0.01),the treatment group was improved more sig-nificant than the control group(P<0.05).After the treatment,the wound healing rate of the treatment group was better than that of the control group compared between two groups(P<0.05).CONCLUSIONS After the interventional vascular surger-y,blood supply state of diabetic foot ulcers patients is improved,but the wound can be further healed through combining Chi-nese medicine wet compress.The effect of the combination treatment is better than interventional vascular surgery only.

OBJECTIVETo develop a new transmission tracking analysis technique during incubation period of respiratory infectious diseases, and to discuss its practical value in the field survey of infectious diseases.

METHODSThe classical epidemiological theory was integrated with geographic information system. The transmission tracking analysis technique was established based on the modeling platform ArcGIS Engine Developer Kit 9.3, using the techniques of address matching, shortest path analysis and buffer analysis, and programming by Visual C++. Eight serious sever acute respiratory syndrome (SARS) cases in Shanghai in year 2003 were then chose as prototype to set up the test cases A-H. The electronic map and population density data were separately collected from Institute of Surveying and Mapping in Shanghai and Shanghai statistical yearbook 2003, to calculate and explore the parameters as length of transmission path, area of buffer zone and key departments by single and multi case analysis module.

RESULTSThe single case transmission tracking analysis showed that the length of transmission track of case A was 129.89 km during April 25th to 29th in 2003, including 12 tracing point and 108 intimate contacts, and the total area of buffer zone was 7.11 km(2) including 81 important institutes, naming 72 schools, 6 kindergartens and 3 gerocomiums. The multi-case transmission tracking analysis showed that the 8 cases shared 5 tracks without any temporal communication. However, there was a spatial communication whose length was 1.42 km and area was 0.60 km(2). There were no important institutes found in this communication area.

CONCLUSIONTransmission tracking technique is practicable and efficient to trace the source of infection, analyze the transmission tracks, establish the isolation buffer area and explore the important geographic positions in epidemiological investigation.

Contact Tracing ; methods ; Disease Transmission, Infectious ; statistics & numerical data ; Epidemiological Monitoring ; Geographic Information Systems ; Humans ; Infectious Disease Incubation Period ; Respiratory Tract Infections ; transmission ; Severe Acute Respiratory Syndrome ; transmission ; Software

Objective To explore the molecular mechanism of pituitary growth hormone adenoma cell proliferation.Methods Functional growth hormone-secreting pituitary adenoma(fGH-PA)tissue samples were collected from 12 patients with acromegaly.The exchange protein 1 directly activated by cAMP(Epac1)mRNA expression levels in fGH-PA tissues and rat RGC-5,MMQ and GH3 cells were determined by qPCR.The expression levels of Epac1 in fGH-PA tissues were determined by immunohistochemistry.Western blot was used to determine the expression levels of Epac1 in MMQ and GH3 cells.Overexpression or knockdown of Epac1,or knockdown of cAMP response element-binding protein(CREB)in GH3 cells,cell cycle changes were determined by flow cytometry,cell proliferation ability was determined by CCK-8 assay,and the expression levels of p-CREB,CREB,Cyclin D1,CDK2 and p21 in cells were determined by Western blot.Results qPCR and immunohistochemistry results showed that the expression levels of Epac1 mRNA and protein in fGH-PA tissues were significantly higher than those in adjacent normal tissues(P<0.05).qPCR and Western blot results showed that compared with RGC-5 cells,the expression levels of Epac1 mRNA and protein in MMQ and GH3 cells were significantly increased(P<0.05).After overexpres-sion of Epac1 in GH3 cells,compared with the Control group,the proportion of cells in G0/G1 phase and S phase in the Epac1-OE group were significantly reduced(P<0.05),and the proportion of cells in G2/M phase were significantly increased(P<0.05);the cell prolifera-tion ability were significantly enhanced(P<0.05);the expression levels of p-CREB and Cyclin D1 in cells were significantly increased(P<0.05),the expression levels of CDK2 and p21 were significantly decreased(P<0.05),while there was no significant change in the expression level of CREB between the two groups(P>0.05).After knockdown of Epac1 or knockdown of CREB in GH3 cells,all of the above results were reversed(P<0.05).Conclusion The overexpression of Epac1 in pituitary growth hormone adenoma cells can up-regulate the levels of p-CREB in cells,and promote adenoma cells to pass through G1/S phase checkpoint,resulting in cell cycle checkpoint disorder and massive proliferation,but it does not affect the expression levels of CREB.

Objective To investigate the characteristics of MRSA colonization and their influence to the prognosis of patients in respiratory intensive care unit (RICU),so as to provide the basis for the caring of the RICU patients.Methods A prospective study included 246 cases of patients admitted to RICU in our hospital during Jan 2010 to Neb 2011.Swab specimens from nares,pharynx,axillae,groin,chest and hands of the patients were collected and cultured at the first day in RICU,and then repeated every 3 days.The patients were divided into MRSA colonization group (n =30) and MRSA non-colonization group (n =216) according to MRSA culture results.Results The MRSA colonization rate was 12.19％ in RICU patients,including 16 cases of hospital-acquired colonization.No significant difference was found in the gender,the APACHE Ⅱ score and APACHE Ⅲ between MRSA colonization group and the MRSA non-colonization group (P ＞ 0.05).The mean age in MRSA colonization group was significantly greater than that of the MRSA non-colonization group [(81.13 ± 6.86) vs (74.78 ± 11.89) ; t =2.955,P =0.003)] ; The number of hospitalization and the mortality in one year of the MRSA colonization group was significantly more than those of MRSA non-colonization group [(1.83 ±1.216) vs (1.06 ±0.257),40.0％ vs 16.7％ ;t =2.998,x2 =9.132;P＜0.01)].MRSA was mainly colonized in nares.Conclusions MRSA colonization has an important impact on the prognosis of RICU patients.More than 50％ of MRSA colonization cases are hospital-associated,suggesting that we should strengthen the bedside care,prevention of cross infection.