BACKGROUND:Chondromodulin-Ⅰ is expressed mainly in the cartilage, but it is little expressed in mesenchymal stem cells. Combined with the previous study of our group, we concluded that chondromodulin-Ⅰmaybe play an important role in inducing mesenchymal stem cells into chondrocytes accurately.
OBJECTIVE:To construct an expression plasmid stably carrying chondromodulin-Ⅰ to up-regulate the expression of chondromodulin-Ⅰ in bone marrow mesenchymal stem cells.
METHODS:Specific primers were designed in rat cartilage for chondromodulin-Ⅰ gene, then the pcDNA3.1 (+) plasmid expression vector was digested by enzyme and directional connected gene to construct pcDNA3.1(+)/ChM-Ⅰ expression vector. Bone marrow mesenchymal stem cells were obtained from rats using the method of density gradient centrifugation combined with adherent culture. Recombinant plasmid pcDNA3.1(+)/ChM-Ⅰ was transfected into rat bone marrow mesenchymal stem cells with liposome method, and G418 selection was used for stable screen of transfected cells. Reverse transcription-PCR and western blot were used to detect chondromodulin-Ⅰ expression in celllines.
RESULTS AND CONCLUSION:The positive clones were digested by enzyme and were identified and sequenced. The results showed that the reality length and sequence of chondromodulin-Ⅰ gene were consistent with the theoretical values, and reading frame was correct. Reverse transcription-PCR and western blot results showed that the expressions of chondromodulin-ⅠmRNA and protein were markedly up-regulated in bone marrow mesenchymal stem cells. Recombinant plasmid pcDNA3.1(+)/ChM-I was successful y constructed, and transfected into rat bone marrow mesenchymal stem cells. After G418 selection, expression of chondromodulin-Ⅰ was up-regulated stably in rat bone marrow mesenchymal stem cells.

Objective To characterize the antibiotic resistance,homology and carbapenemase genotypes of imipenem resistant Acinetobac1ter baumannii isolated from our hospital,and analyze the clonal relatedness of the test strains.Methods Ninety five strains of imipenem resistant A.baumannii were isolated from August 2003 to December 2004 in the First Affiliated Hospital, College of Medicine,Zhejiang University.The MICs of 16 antimicrobial agents against these strains were determined by agar dilution and E-test method.The homology of these isolates was analyzed by pulse-field gel electrophoresis(PFGE).The coding gene of carbapenemases was amplified.PCR products were purified,cloned and sequenced.Plasmid DNA was extracted and purified.Conjugation and Southern blot were performed to locate the position of oxa 23 gene.Results The resistance rates to ampicillin-sulbactam and cefoperazone sulhactam were 67.9% and 30.2%.Polymyxin E had the lowest resistance rate of 17%. The resistance rate to other antimicrobial agents was higher than 90%.The 95 strains,isolated from 10 clinical units,were classified into 6 clones.Clones A and B were predominant clones.All strains produced carbapenemases which were confirmed as OXA 23 by PCR and sequencing analysis.No plasmid was extracted and conjugation was not successful.Southern bolt showed that oxa-23 gene was located on Apal-digested chromosomal segments about 220 kb and 200 kb in Clones A and B,re spectively.Conclusions OXA 23-producing A.baumannii has become one of the most important multi-resistant pathogens in our hospital.Clones A and B have widely spread in our hospital.Oxa-23 gene is located on chromosomal DNA.

Objective:To examine the oncogenic mutations involved in melanoma in Southern China and to provide a theoretical basis for the development of melanoma molecular targeted therapy strategy. Methods:The Sequenom platform (OncoCarta Panel v1.0 and MassARRAY System) was used to determine the prevalence of oncogene mutations in 28 acral melanoma samples, 28 mucosal mel-anoma samples, and 30 non-chronic sun-induced-damage (no-CSD) melanoma samples from Southern China. Results:At least one mu-tation was detected in 33 of the 86 melanomas (38.4%) with mutations observed in BRAF (16.3%), NRAS (10.5%), KIT (5.8%), EGFR (4.7%), HRAS (2.3%), KRAS (2.3%), MET (2.3%), and PIK3CA (1.2%). In BRAF, the age of patients with mutations was significantly lower than those without BRAF mutation (45.7±15.3 vs. 55.9±12.7, P=0.01). Patients with mutations in NRAS were more likely to have ulceration compared with patients without NRAS mutations (88.9%vs. 48.1%, P=0.049). Conclusions:This study represents a compre-hensive and concurrent analysis of the major recurrent oncogenic mutations involved in melanoma cases from Southern China areas. The data have implications for both clinical trial designs and therapeutic strategies.

Objective: To observe the tolerability of Chinese melanoma patients to four-week high-dose interferon alfa-2b(INTRON A(R)，Schering-Plough)therapy. Methods:A total of 29 patients with high risk melanoma[American Joint Committee on Cancer Staging(AJCC)ⅡB-ⅢC]who received adjuvant interferon therapy in our hospital between September 2007 and May 2009 were retrospectively reviewed.Patients received 4 hours of intravenous infusion of interferon alfa-2b fdose range，22.00 million international unit(MIU)to 33.75 MIU]Ⅳ 5 days/week for 4 weeks.The adverse events were evaluated with National Cancer Institute Common Toxicity Criteria(NCI 2.0 version). Results: The average daily dose was 17.63 MIU/(m~2·d).The therapy was ended in two patients because of poor wound healing or intolerability to severe fatigue.The most common adverse events were myelosuppression.Grade 3/4 neutropenia was observed in 69% (20/29)patients and was rapidly reversed after conventional support interventions.Grade 1/2 abnormal hepatic function occurred in 18 cases(62%).Twenty-six patients were followed up for 3 to 22 months.Five patients developed early progression:one with local recurrence，two with regional lymph node metastasis one with in-transit metastasis in the affected limb,and one with distant metastasis. Conclusion: High-dose interferon alfa-2b regimen can be well tolerated by Chinese patients but cannot effectively inhibit subclinical lesions.

Base Sequence ; Binding Sites, Antibody ; Biochemistry ; methods ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Genetic Vectors ; Immunoglobulin Fc Fragments ; genetics ; metabolism ; Immunoglobulin G ; genetics ; metabolism ; Ligands ; Molecular Sequence Data ; Plasmids ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; Staphylococcal Protein A ; genetics ; metabolism

OBJECTIVETo study the rehabilitation effects ergometry on COPD patients.

METHODSThirty COPD out-patients in our Hospital were randomly divided into 2 groups. Rehabilitation group, 15 patients, performed leg ergometry exercise of 80% peak Watt x 30min/d x 3d/w x 12w. Another 15 patients were control group without exercise. All patients received conventional therapy. Pulmonary function testing (PFT), cardiopulmonary exercise testing (CPET), arterial blood gas analysis (ABG), Borg and CAT sores were done at both baseline and 12 w.

RESULTSThere was no statistically difference in lung function testing, blood gas analysis and cardiopulmonary exercise test when pre- exercises between 2 sub-groups. The IC, peak VO2 and peak, W of rehabilitation group significantly increased (P < 0.05); and Borg and CAT.scores significantly decreased (P < 0.05) from baseline; and other PFT and ABG did not change (P > 0.05). While there was no difference in control group (P > 0.05).

CONCLUSIONLeg submaximal ergometry rehabilitation improves health condition and ameliorate dyspnea symptoms in COPD patients.

Blood Gas Analysis ; Dyspnea ; therapy ; Exercise Test ; Exercise Therapy ; Humans ; Pulmonary Disease, Chronic Obstructive ; therapy ; Respiratory Function Tests

OBJECTIVETo study and evaluate the curative effect and mechanism of Qiangxin Granule (QXG) in intervening chronic heart failure (CHF) rats with Xin-qi deficiency complicated blood stasis and edema syndrome (XQD-BS-ES).

METHODSTotally 72 SD rats of clean grade were randomly divided to the normal control group (n =10) and the model group (n = 62). The XQD-BS-ES rat model was established by adriamycin plus propylthiouracil method. Survived modeled rats were then randomly divided to 5 groups i.e., the model group (n = 11, administered with normal saline by gastrogavage), the Western medicine (WM) group (n =11 , administered with perindopril and hydrochlorothiazide by gastrogavage), the low dose QXG (QXG(L)) group (n = 11, administered with 9.26 g/kg QXG by gastrogavage), the middle dose QXG (QXG(M)) group (n = 11, administered with 18.52 g/kg QXG by gastrogavage), the high dose QXG (QXG(H)) group (n = 11, administered with 37.04 g/kg QXG by gastrogavage). After 4 weeks of treatment, left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS), brain natriuretic peptide (BNP), heart rate (HR), respiratory rate (RR), urine output, ear temperature, exhaustive swimming test (EST), tri-iodothyronine (T3), tetra-iodothyronine (T4), thyroid stimulating hormone (TSH), as well as heart, lung, liver weight index and their pathological sections, and high sensitivity C-reactive protein (HS-CRP), angiotensin II (Ang II), carbohydrate antigen 125 (CA125) were detected and compared.

RESULTSCompared with the normal control group, LVEF, LVFS, BNP, HR, RR, urine output, ear temperature, EST, T3, T4, TSH, HS-CRP, Ang II, and CA125 changed significantly in the model group (P < 0.01). Compared with the model group after treatment, LVEF, LVFS, BNP, urine output, EST, T4, heart and liver weight index, HS-CRP, Ang II, CA125 were significantly improved in each QXG group (P < 0.05, P < 0.01). Moreover, TSH was improved in the QXGL and QXG(M) groups (P < 0.05); ear temperature and T3 in the QXG(M) were also improved (P < 0.05); the lung weight index decreased in the QXG(M) and QXG(H) groups (P < 0.01). Compared with the WM group, T4 and CA125 were obviously improved in all QXG groups (P < 0.01); BNP and ear temperature were obviously improved in QXG(L) and QXG(M) groups (P < 0.05, P < 0.01); LVEF, LVFS and TSH were obviously improved in the QXG(M) group (P < 0.05, P < 0.01). And as far as each treatment group, LVEF, LVFS, urine output increased significantly after treatment (P < 0.01); EST obviously increased in QXG(M) and QXG(H) groups (P < 0.01); ear temperature increased in all QXG groups (P < 0.05, P < 0.01). Moreover, compared with the model group, pathological changes of heart, lung, and liver were improved to some degree in each treatment group, especially in the QXG(M) group.

CONCLUSIONSGood curative effect was shown in each QXG group. QXG could improve LVEF, LVFS and BNP of CHF rats of XQD-BS-ES, as well as T3, T4, TSH, EST, urine output, and ear temperature. Moreover, QXG showed superiority than WM group in this respect.

Angiotensin II ; Animals ; C-Reactive Protein ; Chronic Disease ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Edema ; Heart ; Heart Failure ; drug therapy ; Heart Ventricles ; Medicine, Chinese Traditional ; Natriuretic Peptide, Brain ; Qi ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Thyrotropin ; Ventricular Function, Left

OBJECTIVETo establish and evaluate chronic heart failure (CHF) rat model of Xin-qi deficiency complicated blood stasis and edema syndrome (XQD-BSES).

METHODSTotally 40 SD rats were randomly divided into the normal control group (Control), the propylthiouracil (PTU) group, the adriamycin (ADR), and the ADR + PTU group. Normal saline was used as equivalent solvent of each group. Rats in the Control group were intragastrically and intraperitoneally injected with normal saline. Rats in the PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with normal saline. Rats in the ADR group were intragastrically injected with ADR solution and intraperitoneally injected with normal saline. And rats in the ADR + PTU group were intragastrically injected with PTU suspension and intraperitoneally injected with ADR solution. The dose of PTU was 0.2% of daily forage weight, once daily. The dose of ADR was 3.5 mg/kg, once per week. The modeling lasted for 6 weeks. Left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS), brain natriuretic peptide (BNP), heart rate (HR), respiratory rate (RR), urine output, ear temperature, exhaustive swimming test (EST), Tri-iodothyronine (T3), tetra-iodothyronine(T4), thyroid stimulating hormone (TSH) as well as heart, lung, liver weight indices and their pathological sections were integrated and compared.

RESULTSCompared with the Control group, LVEF, LVFS, BNP, HR, RR, heart, lung, liver weight indices, urine output, ear temperature, EST, and T3, T4, and TSH changed significantly in the ADR group, the PTU group, and the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure occurred in pathological sections of heart, lung, and liver. Compared with the ADR group, LVEF, LVFS, BNP, and lung, liver weight indices, urine output, ear temperature, T3, T4, and TSH changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were more serious in pathological sections of heart, lung, and liver. Compared with the PTU group, LVEF, LVFS, BNP, HR, RR, urine output, EST, T4, heart and lung weight indices changed significantly in the ADR + PTU group with statistical significance (P < 0.05), and pathological changes of heart failure were quite serious in pathological sections of heart, lung, and liver.

CONCLUSIONADR + PTU was an appropriate method to establish CHF rat model of XQD-BSES.

Animals ; Edema ; Heart Failure ; diagnosis ; Heart Ventricles ; Humans ; Judgment ; Models, Animal ; Qi ; Rats ; Ventricular Function, Left

Objective To investigate the nutritional status of iodine among residents in Chongqing, and to facilitate scientific prevention and control of iodine deficiency disorders. Methods Select 9 towns in each of the 40 districts (counties) in Chongqing, and collect 40 resident edible salt samples in each of the selected town to detect salt iodine by direct titrimetry. Select 5 towns on the site of the east, west, south, north and middle of every district (county), select 20 children aged 8 to 10 in each of the selected town to collect urine samples and detect urinary iodine by As-Ce catalytic spectrophotometric assay. Results The median of iodine of 14 217 salt specimens by household was 292 mg/kg with a coverage rate of qualified iodized salt of 98.90%( 14 061/14 217). The consumption rate of qualified iodized salt was 95.59%( 13 590/14 217). The median of urinary iodine for 4050 children aged 8 to 10 was 247.20μg/L, of which < 50 μg/L accounted for 4.60%(186/4050), 50-99μg/L accounted for 7.28% (295/4050), 100 - 199 μg/L accounted for 26.44% (1071/4050), 200 - 299 μg/L accounted for 25.58% (1036/4050), 300 μg/L or more, accounted for 36.10% (1462/4050). However, no significant difference was observed between different age groups(x2 = 3.77, P > 0.05). At district (county) level, the median of urinary iodine in 10(25.00%) districts (counties) was 100 - 200 μg/L, that in other 23(57.50%) districts (counties) was 200 - 300 μg/L, and that in other 7(17.50%) districts/counties was greater than 300 μg/L, and statistical significance was observed between different districts/counties (x2 = 441.95, P < 0.01). Conclusions Current iodine nutrition among residents in Chongqing is adequate. While there is excess, need to reduce the amount of salt iodization.

[Objective]To investigate the efficacy and cost analysis ofmicrosurgical clipping and vascular embolization in the treatment of patients with cerebral aneurysms.[Methods]Two hundred and fitly-one patients with cerebral ameurysms,admitted to our hospital from January 2007 to December 2010,were collected in our study;170 patients (174 aneurysms) received microsurgical surgical clipping (clipping group) and other 181 patients (185 aneurysms) were performed vascular embolization (embolization group).Duration of hospitalization,vascular occlusion rate,intraoperative rupture rate,hospital mortality and happening of complications were compared between the 2 groups;and the efficacy were evaluated by modified Rankin scale (mRS) and cost analysis of the 2 treatments were compared 3 months after the surgeries.[Results] The length of hospital stays and cerebral aneurysm rupture rate in patients of the embolization group were significantly shorter/lower than those in the clipping group (P＜ 0.05);the scores of mRS showed no significant differences between the 2 groups (P＞0.05).The cost and sensitivity analysis indicated that the ratio of average cost/good prognosis rate in the clipping group was smaller as compared with that in the embolization group.[Conclusion] The efficacies of microsurgical clipping and embolization in the treatment of cerebral aneurysms are considerable,and both of them have their advantages and disadvantages respectively;however,the microsurgical clipping is cost-effective and economical.