Objective To express the major allergen of Blattella germanica (Bla g 2) in Pichia pastoris and obtain the soluble protein. Methods The known Bla g 2 gene was used to design the primers which had the restriction enzyme sites. PCR method was applied to obtain the Bla g 2 gene. The gene fragment was then cut and ligated with the Pichia expression vector pGAPZaA, resulting in a recombinant plasmid pGAPZaA-Bla g 2. The linearized pGAPZaA-Bla g 2 was transformed into Pichia pastoris GS115 through electroporation, then screened to positive transformants, and the protein was expressed in YPD medium. Purification of the recombinant protein was achieved by metal (Ni2+) chelating affinity chromatography and Western-blotting assay indicated its IgE binding capacity. Results With the expressed reeombinanl protein, SDS-PAGE showed the presence of the product in the supernatant of the culture with Mr 45 000. After 3 days culture, the recombinant protein occupied 50% of the total proteins in the supernatant. The recombinant protein was purified and Western-blot demonstrated an adequate IgE binding capacity of the product. Conclusion A recombinant protein of Bla g 2 has been obtained, which is soluble in the supernatant and therefore can avoid a process of denaturalization and renaturation of the recombinant.

Objective To evaluate the diagnostic value of the diameter ratio of left subclavian artery to aortic isthmus(LSA/AoIS)in fetuses with aortic coarctation(COA).Methods A retrospective study of 79 fetal echocardiographic data was undergone on 49 COA fetuses.The COA cases were divided into four groups according to different gestational week (cases in each group including follow-up review of different gestational weeks):group 1 ,24-27+6 W;group 2,28 -31+6 W;group 3,32 -35+6 W;group 4,36 -39+6 W.The normal fetuses with gestational age matched were also divided into four control groups.The diameter of left subclavian artery(LSA) and aortic isthmus (AoIS) were measured,the ratio of LSA/AoIS of each group were calculated,and the data between the COA and control groups were compared.ROC curve analysis was used for LSA/AoIS to predict the demarcation point of postnatal COA.Results ①There was no statistic difference in the LSA diameter between COA groups and control groups (P >0.05):group 1 (1 .87±0.42)mm vs (1 .75 ±0.25)mm;group 2 (2.25 ±0.36)mm vs (2.21 ±0.22)mm;group 3 (2.74±0.32)mm vs (2.90 ±0.29)mm;group 4 (2.83 ±0.28)mm vs (3.06 ±0.30)mm;②The ratio of LSA/AoIS was significantly increased in COA groups than those in control groups:group 1 (0.88± 0.15) vs (0.66±0.06),group 2 (0.85±0.13)vs(0.64±0.05),group 3 (0.94±0.17)vs(0.73±0.07), group 4 (0.94±0.18)vs(0.70±0.07),the difference was statistically significant(P <0.05);③The cut-off value of LSA/AoIS ratio were 0.78,0.73,0.83,0.78 in each group,respectively.Conclusions The ratio of LSA/AoIS can be used as a useful echocardiographic parameter which suggest the presence of aortic coarctation.This ratio≥0.8 may have a certain diagnostic significance.

Objective To investigate the difference in cardiac structure and hemodynamics characteristics between fetuses with isolated persistent left superior vena cava (LSVC)and normal fetuses by using fetal echocardiography,in order to explore whether isolated PLSVC has some influence on the development of fetal heart structure development.Methods Twenty-two fetuses diagnosed with isolated PLSVC in the second and third trimester were enrolled in this study.The PLSVC fetuses were divided into two groups:group 1 ,twenty-two fetuses in the second trimester (23-27+6 W);group 2,twenty fetuses in the third trimester (28-39+6 W).Forty-nine fetuses without intra-cardiac and extra-cardiac anomalies with gestation age matched were involved as normal control groups:25 fetuses in the second trimester,and 24 fetuses in the third trimester.The parameters of cardiac structure and hemodynamics of the four groups were measured,including left and right sideheart size,the diameter of foramen ovale(FO),aorta(AO),aortic isthmus and pulmonary artery (PA).The ratio of RV/LV,RA/LA and PA/AO were calculated.The hemodynamic parameters included the flow velocity across mitral valve(MV),tricuspid valve(TV)and FO. All the parameters of PLSVC groups were compared with the normal groups respectively.Results In the second trimester group,the ratio of RV/LV and PA/AO of the PLSVC fetuses were significantly larger than normal,while the AO diameter,aortic isthmus diameter were smaller than normal group (P <0.05). However in the third trimester group,only the ratio of PA/AO of the PLSVC group was significantly greater,and the aortic isthmus diameter was still smaller than the normal group (P <0.05).Conclusions Isolated PLSVC connected with coronary sinus is associated with structural differences from normal.A dilated coronary sinus may have an influence on the development of fetal left heart structures.

BACKGROUND: Both animal experiments and clinical practice have confirmed that mild or moderate hypothermia is effective in reducing secondary brain injury, but its effect on homeostasis is not very clear.OBJECTIVE: To investigate the effect of a combined therapy of mild hypothermia and hibernation on the homeostasis of patients with severe brain injury.DESIGN: A randomized controlled study.SETTING: Neurosurgical Institute of Beijing; Neurosurgical Department of the First Clinical Medical College Affiliated to Harbin Medical University;and Neurological Department of the Second Clinical Medical College Affiliated to Harbin Medical University.PARTICIPANTS: The study was conducted at the Department of Neurosurgery, the First Affiliated Hospital of Harbin Medical University, from June to December 2002. Totally 24 patients (aged 35-60 years) with severe cerebral hemorrhage or brain injury were randomly divided into combined therapy group and normothermia group. Their Glasgow Coma Scale scores ranged from 3 to 8. The subjects signed the informed consent.METHODS: Within 10 hours of their injury, patients in hypothermia and hibernation combination group were given half dosage of No. 1 hibernation cocktail (chlorpromazine 25 mg, pethidine hydrochloride 50 mg, and promethazine 25 mg), and were cooled by cooling blankets to make their body temperature dropped to 32-34 ℃ (rectal temperature). Their temperature was kept within this range for 5 days, at 35 ℃ for 24 hours, and then was slowly increased to their normal level. The body temperature of patients in normothermia group was maintained at 37-38 ℃. The mean arterial pressure and heart rate of all patients were measured continuously by HP monitor. On the 3"d and 7th days of hospitalization, intracranial pressure and creatine phosphate kinase were measured via lumbar puncture.Femoral artery puncture was performed to check the partial pressure of arterial O2 and CO2. Platelets count and blood electrolytes K+ and Na+ concentration of each patient were measured, too. On the 7th day Glasgow Outcome Scale scores of each patient and mortality of each group were recorded.activity of creatine phosphokinase, platelets count, blood K+ and Na+ conand CO2 of patients in combined therapy group on the 3rd and 7th days of hospitalization.intracranial pressure, creatine phosphokinase and platelets count: The decreased values of intracranial pressure, creatine phosphokinase and platelet number in combined therapy group were all significantly higher than those in normothermia group [(104.09±54.90), (58.75±25.33) mm H2O; (26.95±19.22), (10.17±7.18) μkat/L; (89.82±46.36)×109/L, (48.83±44.59)×109/L,the mean arterial pressure, blood electrolytes, and partial pressure of artewas significantly lower than that of normothermia group (25.0%, 66.6%,P ＜0.05).CONCLUSION: This combined therapy of hypothermia and hibernation can effectively decrease intracranial pressure and creatine phosphokinase,but has no significant effects on the mean arterial pressure, blood electrolytes concentration, and partial pressure of arterial O2 and CO2. It has the risk of disturbing the patients' hematopoiesis.

Objective To explore the incidence of G-6-PD deficiency in Dongguan District,Dongguan,Guangdong province. Methods Cord blood was obtained in 9676 cases of live births. G6PD/6PGD ratio was measured. Results Two hundred and sixty-five cases were diagnosed as G-6-PD deficiency giving an incidence of 2.74%. The incidence in male (4.07%,230/5646) was significantly higher than that in female (0.875,35/4030) ( P

Humans ; Pneumoconiosis ; therapy ; Self Care

Objective To discuss the drug resistance and the vicissitude of Pseudomonas aerugi-nosa in Intensive Care Unit (ICU), in order to provide basis for rational use of antibiotics in clinic. Methods Identification of bacteria was performed by applying ATB Expression Identification System produced by BioMerieux Company; drug susceptibility tesr was done by using K-B method. The drug resistance of Pseudomonas aeruginosa was compared between ICU and general wards; and then the difference of drug resistance was investigated between 2006 and 2007. Resnlts The more sensitive an-tibiotics to Pseudomonas aeruginosa in ICU of our hospital were as follows, amikacin (resistance rate 6.7%-9.4%), eiprofloxacin (resistance rate 7.7%-24.7%), cefoperazone/sulbactam (resistance rate 11.5%-34%), ceftazidime (resistance rate 11.5%-37.7%), aztreonam (resistance rate 20.2%-40.6%). The drug resistance rate of ICU was higher than that of the general wards, and the drug re-sistance rates to ciprofloxacin, ceftazidime, cefoperazone/sutbaetam,meropenem, imipenem and aztreo-nam in 2007 were significantly higher than those in 2006 (P<0.01). Conclusion The drug resistance rate of Pseudomonas aeruginosa in ICU of our hospital was very high,and it is rising year by year. So it is important to retrospect the results of the drug resistance periodically and provide the up-to-the-mi-nute data of epidemiology and vicissitude of drug resistance.

Objective To observe the effect of Danhong Injection(DI)on the thyroid hormone level in patients with angina due to coronary heart disease(CHD).Methods Sixty CHD angina patients were equally randomized into the treatment group and the control group.The two groups received routine western medicine such as aspirin,angiotensin convert enzyme inhibitor(ACEI),statins,beta-blockers and nitrate esters,and the treatment group was additionally given intravenous drip transfusion of DI 20 mL,qd.Twenty days constituted one treatment course.After treatment,the therapeutic effect was evaluated and the changes of serum thyroid hormone level were observed.Results In the control group,13 patients were markedly effective,9 effective,8 ineffective,and the total effective rate was 73.3%;in the treatment group,19 patients were markedly effective,6 effective,5 ineffective,and the total effective rate was 83.3%;the difference between the two groups was significant(P

Objective To study the chemical constituents from the root of Rumex gmelini.MethodsThe compounds were isolated and purified by chromatography,polyamide column chromatography,and preparation HPLC etc.Their structures were elucidated by physicochemical and spectroscopic evidences.Results Ten compounds were identified as:nepodin (Ⅰ),emodin (Ⅱ),citreorosein (Ⅲ),chrysophanol 8-O-?-(6'-acetyl) glucopyranoside (Ⅳ),chrysophanol 8-O-?-D-glucopyranoside (Ⅴ),resveratrol (Ⅵ),9,9'-dianthranone-2,2'-dimethyl-5,5'-bis (?-D-glucopyranose)-9,9',10,10'-tetrahydro-4,4'-dihydroxy-10,10'-dioxo (trivial name:rumoside A) (Ⅶ),emodin-8-O-?-D-glucopyranoside (Ⅷ),resveratrol-3-O-?-D-glucoside (Ⅸ),and rutin (Ⅹ).Conclusion Compound Ⅶ is a new compound,named rumoside A.Compounds Ⅴ,Ⅷ,and Ⅹ are separated from R.gmelini for the first time.Compounds Ⅲ and Ⅳ are the compounds which have been found in the plants of Rumex L.for the first time.

ObjectiveTo explore environmental conditions under which bone marrow stromal cells could be induced into osteogenic phenotype.MethodsRat bone marrow stromal cells were isolated and proliferated in vitro, and the 3rd passage was divided into the group A (control group), group B (cells cultured in the medium containing dexamethasone, β-glycerol disodium phosphate salt hydrate, vitamin C and active form of vitamin D3), and group C (on the bases of group B, the cells were cultured additionally with fracture hematoma extract). On the post-induction day 5, 8, and 11, the morphological changes were observed and the osteogenic markers such as alkaline phosphotase (ALP), collagen type Ⅰ (Col Ⅰ) and osteocalcin (OCN) were assayed with immunohistochemical staining, the calcification was manifested with von Kossa staining.ResultsIn the group A, no evident osteogenic effects had been observed. In the group B, on 5th day post-induction, some bone marrow stromal cells underwent a morphological change, and mild expression of ALP and Col Ⅰ was observed but with no calcification effect. On 8th day post-induction, the ratio of morphologically changed cells increased, and the expression of ALP and Col Ⅰ increased still with no evident calcification. On 11th day post-induction, anti-OCN staining was positive and the calcium nodes were showed by von Kossa staining. The phenotype changes in the group C were similar to group B, but were more evident.ConclusionBone marrow stromal cells can be induced into osteogenic phenotype in vitro with small molecular inducers. Fracture hematoma extract can enhance this effect thus might be used as an addictive in osteogeneration.