AIM: To investigate the use of therapy ultrasound to enhance nonviral gene delivery. METHODS: Endothelial cells (EC) and vascular smooth muscle cells (VSMC) were cultured in 6-well plates. Plasmid (pcDNA3.1/His/LacZ) with or without microbubbles at the different concentrations was transfected into the cells with the use of ultrasound for 1 min at 2 MHz, 1.8 mechanical index (MI). Additional controls included ultrasound alone, microbubble alone and microbubble plus plasmid. The rate of blue cells and the activities of ?-Gal were measured. In addition, cell viability was detected with different time from 1 to 30 min of ultrasound irradiation and the different concentrations of microbubbles. RESULTS: In the group of ultrasound with microbubble, the rate of blue cells and activity of ?-Gal markedly increased by 60% and 9-fold, respectively. Microbubbles at concentration of 10% led to the highest transfection effect. Ultrasoud exposure at 1 to 30 minute had no cell toxic effects, while microbubbles at the concentration of 50% had significant effect on cell survival. CONCLUSIONS: Albumin-coated microbubbles markedly enhance gene delivery by therapeutic ultrasound-mediated microbubble destruction, which can be used as a safe and practicality vectors in gene therapy.

Objective: To investigate the impact and its possible mechanism of hydrogen sulfide (H2S) on myocardial collagen remodeling in experimental rats with diabetic mellitus (DM).
Methods: Rat’s DM model was established by intraperitoneal injection of STZ at 40 mg/kg. A total of 40 SD rats were randomly divided into 4 groups:Control group, DM group, DM+NaHS group, in which NaHS worked as exogenous donor of H2S and NaHS control group. n=10 in each group, all animals were treated for 8 weeks. The cardiac collagen deposition was observed by Masson staining, protein expressions of cardiac collagen types I, III, IV and transforming growth factorβ1 (TGF-β1), connective tissue growth factor (CTGF) were examined by Western blot analysis.
Results: Compared with Control group, DM group showed increased protein expressions of cardiac collagen types I and III, up-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups. Compared with DM group, DM+NaHS group presented reduced cardiac collagen expression, decreased expression of collagen types I and III, down-regulated expressions of TGF-β1 and CTGF, P<0.05;while the expressions of collagen type IV were similar between 2 groups.
Conclusion: H2S may improve the myocardial collagen remodeling in experimental DM rats, the mechanism might be related to the down-regulation of TGF-β1 and CTGF expression.

OBJECTIVETo provide evidence for intervention measures on safe injection through a survey on the safety of injection in the health facilities in Wulong county, Chongqing city, China.

METHODSAn investigation on injection safety was conducted among health care providers with a uniform questionnaire.

RESULTSGlass syringes which were not appropriately sterilized was found in 52.2% (12/23) of the health facilities. Injection practices were not correct in 31.4% (11/35) of the health facilities. Used disposable syringes were discarded directly without any disposal in 63.6% (21/33) of the health facilities. At least one of the unsafe injection practices mentioned above existed in 77.1% (27/35) of the health facilities.

CONCLUSIONSThe main unsafe factors in the practice of injection are included: the practice to change needle but not syringe in penicillin allergic test and the injuries caused by used disposable syringes in medical staff. The important measures to improve injection safety were to train the medical staff on the knowledge about safe injection and the risks caused by unsafe injection, and to equip the health facilities with safe tools to destroy the used disposable syringes.

Adult ; China ; epidemiology ; Cross-Sectional Studies ; Disinfection ; statistics & numerical data ; Disposable Equipment ; statistics & numerical data ; Equipment Reuse ; statistics & numerical data ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Injections ; adverse effects ; statistics & numerical data ; Male ; Risk Factors ; Rural Health Services ; Safety ; Sampling Studies ; Surveys and Questionnaires ; Syringes

OBJECTIVEAn epidemic of severe acute respiratory syndrome (SARS) hit Beijing, China, between March and July 2003 with an attack rate of 1.9 per 10 thousand. (2,521 cases). To control the epidemic of SARS, a total of 30,173 residents were quarantined either in their residence or in quarantine sites. In order to understand the personal need of being quarantined and to estimate the risk of developing SARS during the quarantine period, a survey on the quarantined residents of Haidian District, Beijing, China was carried out.

METHODS33 precincts in Haidian District divided into five groups (7 in north, 6 in south, 7 in west, 6 in east and 7 in central of Haidian District) according to the location of the precincts were involved. The director of Center of Disease Control and Prevention of Haidian District was asked to select 1 precinct from each group according to the workload of the precinct quarantine officers. From those 5 precincts we obtained lists of all quarantined persons from the precinct quarantine officers. All quarantinees were asked to complete a self-administered questionnaire. The SARS patients were diagnosed and verified according to the diagnosis criteria released by Chinese Ministry of Health which was equivalent to the SARS 'probable case' definition of WHO. All SARS cases had been reported to the relative authorities.

RESULTSBy May 23, 2003, 5,186 persons had been quarantined in Haidian district, accounting for 0.23% of all residents. 1,028 of sampled quarantined residents completed the questionnaire. Of those who completed the questionnaire, 2.3% (95% CI: 1.5% - 3.5%) developed SARS while under quarantine. The median quarantine period was 14 days (range 1 day to 28 days). 61% of the quarantinees had a direct contact history with SARS patients, and all secondary SARS patients developed through contact to these quarantinees. The remaining 39% quarantinees who did not have a direct contact history with SARS patients had not developed SARS during the period under quarantine. 37% of the quarantees had direct contact during original patients' symptomatic period with an AR of 6.3%. Persons who looked after the illed SARS patient(s) during their symptomatic period, had an highest attack rate of 31% (95% CI: 20% - 44%). 63% (95% CI: 60% - 66%) of the total quarantined persons did not have direct contact with a SARS patient during his/her symptomatic period, with an attack rate of 0% (95% CI: 0% - 0.73%).

CONCLUSIONOnly those persons having direct contact with ill SARS patients need to be under quarantine. The overall cost for quarantine on SARS prevention could be reduced by as much as 63% if the quarantine program was limited to this group. No evidence was found that SARS patient was infective during the incubation period.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; prevention & control ; Female ; Humans ; Infant ; Male ; Middle Aged ; Quarantine ; methods ; organization & administration ; psychology ; Risk Factors ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; psychology ; Time Factors

OBJECTIVETo evaluate the infectivity of severe acute respiratory syndrome (SARS) during its incubation period by investigating chains of transmission and individuals isolated for medical observation with a view to providing scientific evidence for updating protocols of medical isolation.

METHODSIndividuals related with the two SARS chains of transmission in Beijing in 2003 and a group of individuals isolated for medical observation in Haidian district of Beijing during the SARS outbreak were selected as subjects of study. Contactors with SARS patients and those with symptom development following the contacts were investigated via questionnaire. Serum samples were collected from super transmitters and tested for SARS-CoV antibody by neutralization test and enzyme linked immunosorbent assay (ELISA).

RESULTSA total of 1112 contactors were investigated in three surveys. Of them, 669 had a history of close contact with symptomatic SARS patients, 101 developed symptoms with a rate of 15.1%, 363 had a history of close contact with patients in their incubation period, none of whom developed symptoms (0%). Serum samples were collected from 32 highly-exposed individuals, of whom 13 developing SARS symptoms after contact had serum samples positive for SARS-CoV antibody. Samples collected from the asymptomatic contactors were all negative for SARS-CoV antibody.

CONCLUSIONSARS cases are infectious only during their symptomatic period and are non-infectious during the incubation period. Isolation for medical observation should be placed for individuals who are in close contact with symptomatic SARS patients. The results of our study are of decisive significance for the Ministry of Health to the definition of SARS close contactor.

China ; epidemiology ; Disease Outbreaks ; Humans ; Infectious Disease Incubation Period ; SARS Virus ; physiology ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology

Objective To analyze the characteristics of clinical manifestations, risk factors, therapies and acute outcomes in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage. Methods Seventy-five patients with cerebral venous sinus thrombosis were included in the study. According to the radiological findings on the brain image, patients were divided into two subgroups:cerebral hemorrhage group and non-hemorrhage group. The demo?graphic data, potential risk factors, clinical manifestations, radiological features, therapeutic strategies and acute out?comes were compared between two subgroups, and high risk factors were also analyzed. Results There were seventy-five patients with cerebral venous sinus thrombosis in the present study. Twenty-eight patients of them (37.2%) had cerebral hemorrhage whereas the remaining forty-seven patients (62.7%) did not have cerebral hemorrhage. Pregnancy/puerperi?um were significantly higher in patients with cerebral hemorrhage (with vs without;28.6%vs. 6.4%, P=0.015), while in?fection was markedly higher in patients without cerebral hemorrhage (with vs without;7.1% vs. 29.8%, P=0.021). Head?ache (92.9% vs. 70.2%, P=0.021), unconsciousness (25.0% vs. 6.4%,P=0.034), seizures (53.6% vs. 19.1%, P=0.002) and motor deficits (35.7% vs. 12.8%, P=0.019) were more common in patients with cerebral hemorrhage. Moreover, mul?tiple sinus involvement (1.4% vs. 44.7%, P=0.024) was significantly higher and the acute outcomes(mRS≥3: 46.4%vs.17.0%, P=0.006)were poorer in patients with cerebral hemorrhage. Binary Logistic analysis showed that pregnancy/pu?erperium (P=0.004) and multiple sinus involvement were positively, whereas infection was negatively correlated with cere?bral venous sinus thrombosis and hemorrhage ( P=0.007;P=0.03). Conclusions Pregnancy/puerperium, headache, uncon?sciousness, seizures, motor deficits and multiple sinus involvement are more frequently in patients with cerebral venous sinus thrombosis and hemorrhage, and the acute outcomes are poorer in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage.

OBJECTIVETo establish the determination method for complanatoside A in seeds of Astragalus complanatus.

METHODAn HPLC method has been developed to separate complanatoside A on ZORBAX EXTEND-C18 (4.6 mm x 250 mm, 5 microm) column with acetonitrile-water-phosphoric acid (20:80:0.2) as mobile phase and UV detection at 267 nm.

RESULTThe good linearity of complanatoside A ranged 0.086-0.430 microg, r = 0.9999. An average recovery of 99.8% (n = 5) was obtained with a RSD of 1.0%.

CONCLUSIONThe established method is proved to be stability, fast, accurate and can be used for quantification of Complanatoside A in Semen Astragali Complanati.

Astragalus Plant ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Flavonols ; analysis ; Glucosides ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Seeds ; chemistry

OBJECTIVETo establish a high-performance liquid chromatography-mass spectrometry (LC/MS)-based method for efficient determination of melamine in urinary calculi in children taking melamine-contaminated formula milk powder.

METHODSLC/MS was employed to determine the contents of melamine in urinary calculi surgically removed from 17 children with a history of taking melamine-contaminated milk powder and in 4 samples of uric acid stone from adults. The positive ionization mode of electro-spray ionization source was used, and the limit of melamine determination was 0.1 mg/kg.

RESULTSNo melamine was detected in the 4 uric acid stone samples from adults. Melamine was detected in 4 samples of urinary calculi from the 17 children, with the concentration ranging from 0.8 to 64 mg/lkg.

CONCLUSIONLC/MS is simple and effective for detecting melamine in urinary calculi, which is helpful to the treatment and follow-up.

Chromatography, High Pressure Liquid ; methods ; Dairy Products ; adverse effects ; Female ; Food Contamination ; Humans ; Infant ; Male ; Mass Spectrometry ; methods ; Triazines ; analysis ; Urinary Calculi ; chemically induced ; chemistry

BACKGROUNDA high mortality rate of pancreatic cancer becomes a bottleneck for further treatment with long-term efficacy. It is urgent to find a new mean to predict the early onset of pancreatic cancer accurately. The authors hypothesized that genetic variants of cationic trypsinogen (PRSS1) gene could affect trypsin expression/function and result in abnormal activation of protease activated receptor-2 (PAR-2), then lead to pancreatic cancer. The aim of this study was to elaborate some novel mutations of PRSS1 gene in the patients with pancreatic cancer.

METHODSTotally 156 patients with pancreatic cancer and 220 unrelated individuals as controls were enrolled in this study. The mutations of PRSS1 gene were analyzed by direct sequencing. K-ras Mutation Detection Kit was used to find the general k-ras gene disorder in the pancreatic cancer tissue. Then the clinical data were collected and analyzed simultaneously.

RESULTSThere were two patients who carried novel mutations which was IVS 3 + 157 G > C of PRSS1 gene in peripheral blood specimens and pancreatic cancer tissue. What's more, it was surprising to find a novel complicated mutation of exon 3 in PRSS1 gene (c.409 A > G and c.416 C > T) in another young patient. The complicated mutation made No. 135 and No. 137 amino acid transfer from Thr to Ala and Thr to Met respectively. No any mutation was found in the normal controls while no mutations of k-ras gene were detected in the three patients.

CONCLUSIONMutations of PRSS1 gene may be an important factor of pancreatic cancer.

Adult ; Asian Continental Ancestry Group ; Female ; Humans ; Male ; Mutation ; Pancreatic Neoplasms ; genetics ; Trypsin ; genetics

OBJECTIVETo explore the effects of hydrogen sulfide (H(2)S) on myocardial fibrosis and expressions of MAPK1/3 and MMP-8 in diabetic rats.

METHODSForty adult male SD rats were randomized into 4 groups, namely the control group, diabetes mellitus group (STZ group), diabetes mellitus with H(2)S treatment group (STZ+H(2)S group), and normal rats with H(2)S treatment group (H(2)S group). Diabetes was induced by intraperitoneal injections of 40 mg/kg streptozotocin (STZ). The rats in the control group received daily intraperitoneal injections of saline, and those in STZ+H(2)S group and H(2)S group were given NaHS (100 µmol/kg) injections. After 8 weeks, the pathologies of cardiac fibrosis were examined with HE staining, and the expressions of collagen I, MAPK1/3 and MMP-8 were analyzed with Western blotting.

RESULTSCompared with the control group, the diabetic rats showed increased collagen content and obvious interstitial fibrosis in the myocardial tissue with significantly increased expression levels of collagen I, MAPK1/3 and MMP-8 (P<0.05); all these changes were obviously reversed by treatment with H(2)S (P<0.05). Collagen I, MAPK1/3 and MMP-8 expression levels and the degree of myocardial fibrosis were comparable between H(2)S group and control group (P>0.05).

CONCLUSIONHydrogen sulfide can attenuate cardiac fibrosis in diabetic rats, and the mechanism may involve the inhibition of MAPK1/3/MMP-8 signal pathway.

Animals ; Collagen Type I ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Fibrosis ; Hydrogen Sulfide ; pharmacology ; Injections, Intraperitoneal ; Male ; Matrix Metalloproteinase 8 ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley