Construction and evaluation of a pUC-type prokaryotic promoter reporter system based on lacZ gene / 生物工程学报

To construct a prokaryotic promoter report system with wide applicability, a series of pFGH reporter vectors based on lacZ gene and pUC replicon were constructed from plasmid pFLX107 through the replacement of multiple cloning sites and sequence modifications. The plasmid with the lowest background activity was selected as the final report system with the lacZ gene deletion strain MC4100 as the host bacterium, following by testing with inducible promoter araBAD and the constitutive promoter rpsM. The background activity of pFGH06 was significantly lower than that of other plasmids of the same series, and even lower than that of reference plasmid pRCL at 28 °C (P<0.01). Further evaluation tests show that the plasmid pFGH06 could be used to clone and determine the activity of inducible promoter or constitutive promoter, and the complete recognition of the target promoter could be achieved through blue-white selection in the simulation test of promoter screening. Compared with the reported prokaryotic promoter report systems, pFGH06 has the advantages of smaller size, more multiple clone sites, adjustable background activity, high efficiency of promoter screening and recognition, thus with a wide application prospect.

Clinical significance of NT-proBNP in evaluation of cardiac function in patients with severe burn / 实用医学杂志

Objective To explore the clinical significance of N-terminal pro-brain natriuretic peptide(NT-proBNP)in the evaluation of cardiac function in patients with severe burns. Methods A total of 78 patients with severe burns in intensive care unit(ICU)in our hospital were selected. Patients were divided into a heart failure group(n = 31)and a non-heart failure group(n = 47)according to the heart failure complication condition. The changes of NT-proBNP level in different time ,heart function index ,relevant index and mortality were compared between the two groups. Results The levels of NT-proBNP ,troponin(CTnT),creatine kinase(CK),creatine kinase isoenzyme (CK-MB),stroke volume index (SVI),cardiac index (CI) and central venous pressure (CVP),and the score of acute physiologic and chronic health conditions(APACHE II)in the heart failure group were shown to be significantly increased than those of the non-heart failure group (P < 0.05). The mean arterial pressure(MAP)in the heart failure group was significantly lower than the non-heart failure group(P < 0.05). Mortality rate in the heart failure group was shown to be significantly higher than the non-heart failure group (P < 0.05). Conclusion Heart failure is a common complication in patients with severe burns. The mortality rate in the population with heart failure is relatively high ,for which NT-proBNP can effectively assess the conditions and prognosis.

Establishment of a diet-induced obesity model in zebrafish larvae / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a diet-induced obesity model in zebrafish larvae.</p><p><b>METHODS</b>At 7 days post-fertilization (dpf), 200 zebrafish larvae with normal development were randomly allocated to two groups with the feeding quantity of 30 mg per day (normal feeding group) or 180 mg per day (overfed group) for 20 days. The weight, length, BMI, triglyceride (TG) and total cholesterol (TCH) of each group were measured. Whole-mount Oil Red O staining, frozen Oil Red O staining and hematoxylin-eosin (HE) staining were used to estimate the rate of hepatic steatosis and liver histology of the zebrafish. The dynamic change of hepatic lipid droplets and distribution of adipose tissue were observed with Nile Red staining in overfed zebrafish in vivo.</p><p><b>RESULTS</b>The weight, length, BMI and TG of overfed zebrafish were significantly increased compared with those in normal feeding group. Whole-mount Oil Red O staining showed that the percent of hepatic steatosis in overfed group (89.4%) was markedly higher than that in normal feeding group (20.7%). Macrovesicular steatosis was observed in the liver of the overfed larvae. Nile Red staining visualized hepatic lipid droplets and the distribution of larval adipose tissue, which increased with feeding time in the overfed zebrafish. Starving larvae showed depletion of fat and hepatic lipid, and adipose tissue was induced after refeeding.</p><p><b>CONCLUSIONS</b>We successfully established an diet-induced obesity model in zebrafish larva, in which Nile Red staining allows in vivo observation of the adipocytes and hepatic lipid droplets.</p>

Study on the clinic application value of ultrasonography examination of optic nerve sheath diameter in brain injury / 中国医师杂志

Objective To explore the clinic application value of ultrasonography examination of optic nerve sheath diameter(ONSD) in brain injury.Methods From July 2008-June 2011,90 cases of brain injured patients were chosen as experimental group including light (A group),medium (B group),and heavy (C group) brain injured patients according to the admission GCS score ;50 cases of conventional physical examination and 90 cases of volunteers 50 in neurosurgical outpatient were chosen as control group.The ONSD of both groups were measured 3 mm behind the globe through orbital using color sonographic with different time after admission.3 times measurements were carried out for every optic nerve sheath.All client's ONSD mean and standard deviation were calculated.In 0.5 h after color dopplar ultrasound examination,lumbar vertebra puncturing measured intracranial pressure in different groups.Results After admission (1d,3 d,7 d,14 d),the ONSD of A group was (4.54 ±0.32)mm,(4.42 ±0.30)mm,(4.44 ±0.32) m,and (4.43 ± 0.25) mm,respectively; The ONSD of B groups was (4.48 ± 0.28) mm,(4.52 ± 0.24) mm,(4.46 ±0.28)mm,and (4.38 ±0.22)mm,respectively; The ONSD of C group was (5.67 ±0.35)mm,(6.36 ± 0.42) mm,(5.65 ± 0.23) mm,and (4.76 ± 0.35) mm,respectively.After admission (1 d,3 d,7 d,14 d),the intracranial pressure (IP) of A group was (82 ± 11) mmH2O,(79 ± 12) mmH2O,(90 ±15) mmH2O,and (86 ± 14) mmH2O,respectively; The IP of B group was (78 ± 15) mmH2O,(85 ± 10)mmH2O,(78 ± 16) mmH2O,(80 ± 11) mmH2O,The IP of C group was (225 ± 26) mmH2 O,(288 ± 23)mmH2O,(256 ± 23) mmH2O,(122 ± 18) mmH2O,respectively.Group D had the ONSD average of (4.58± 0.41)mm and IP of (88 ± 10)mmH2O after eyeball 3-mm place.No difference was found between A and B,A and D,or B and D (P＞0.05) ; A difference was found between A and C,B and C,or D and C (t =12.24～24.67,P＜0.01).Conclusions The ONSD and IP in light medium brain injured patients had no change.In patients with severe brain injury,IP changed with the time after injury,the ONSD increased with the IP,the ultrasonography examination of ONSD with the important value in the diagnosis and treatment can respond the IP increase,which is a non-invasion,convenient,fast,and feasible method for evaluation of cranial high pressure.

Questionnaire survey on obstructive sleep apnea hypoventilation syndrome and related complications / 中华全科医师杂志

Objective To investigate relationship between obstructive sleep apnea hypoventilation syndrome(OSAHS) and hypertension.Methods Questionnaire survey was conducted among 1573 local permanent residents in Guangzhou during March and June 2006 to understand prevalence of hypertension and snoring,and polysomnography monitoring was performed for 347 of them with moderate and severe snoring to estimate prevalence of OSAHS.Risk factors for hypertension was screened with multivariate logistic regression analysis.Results In total,1547 copies of valid questionnaires were collected,and 184 cases (11.9%) of hypertension were found with a prevalence of 10Department of Endocrinology,Shanxi Provincial People's Hospital,Taiyuan 030012,China 0%,49 of diabetes with a prevalence of 2.7%,and 212 of dyslipidemia with a prevalence of 10.4%.Prevalence of OSAHS was found in 34 of them (2.2% ) with Epworth sleepiness scale (ESS) score equal to or greater than nine.Prevalence of hypertension in OSAHS group was 32.5%,much higher than that in non-snoring group (8.3% ) and snoring group ( 17.1% ).Results of logistic regression analysis showed that age,gender,OSAHS,diabetes and dyslipidemia all were high-risk factors for hypertension,with OSAHS ranking the third.Conclusion OSAHS is one of high-risk factors for hypertension.

Sodium Channel Current In Hypertrophied Myocardiocytes in Spontaneously hypertensive Rat / 中华高血压杂志

Objective To study the change in sodium channel current(ⅠNa)in the developement of the left ventricular hypertrophy in spontaneously hypertensive rat(SHR).Methods ⅠNa and membrane capacitance(MC)were measured in myocytes of 10,24 and 34-week SHR by the whole-cell patch-clamp technique.Systolic blood pressure(SBP)and left ventricular mass index(LVMI)were detemined.10-week old Wistar rats served as controls.Results LVMI and MC of SHR were significantly higher than those of Wistar rat(P0.05),however the density of ⅠNain 34-week SHR were higher than those of Wistar rat(-18.3?1.9)vs(-15.3?2.0)pA/pF,P

Roles of Rho-kinase in rat cardiac fibroblasts proliferation and collagen synthesis induced by angiotensin Ⅱ / 中国病理生理杂志

AIM: To investigate the role of Rho-kinase signal pathway in rat cardiac fibroblasts (CFBs) proliferation and collagen synthesis induced by angiotensinⅡ (AngⅡ). METHODS: CFBs of neonatal Sprague-Dawley (SD) rats were isolated with the method of trypsin digestion and differential anchoring velocity. The CFBs were stimulated with AngⅡto induce fibrosis. Proliferation of CFBs was observed by MTT coloricmetric assay. Synthesis of collagen was detected by the hydroxyproline. The expression of Rho-kinase mRNA was examined using RT-PCR analysis. The extent of phosphorylation of myosin-binding subunit (MBS-P) of myosin phosphatase was quantified by Western blotting analysis, which was used to evaluate the activity of Rho-kinase.RESULTS: (1) Stimulation of neonatal SD rat CFBs with AngⅡ (10-7 mol/L) significantly increased CFBs proliferation and collagen synthesis (P