Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

?AlM: To evaluate the clinical effects and security of posterior chamber implantable Collamer lens ( lCL ) implantation in patients with extreme highly myopia.
?METHODS:ln this study, 18 patients ( 32 eyes ) with extreme highly myopic patients who had undergone posterior chamber lCLs implantation from July 2010 to July 2013 were evaluated. Diopter -10. 5 ~ 19. 0D, and astigmia -0. 5 ~4. 5DC. Changes in intraocular pressure ( lOP ) , refraction, visual acuity and corneal endothelium, anterior chamber depth, iris, high arch, lens were noted at 1d, 1wk, 1, 3mo and 1a after surgery respectively, and follow-up was of 1a.
? RESULTS: Before surgery, the uncorrected visual acuity (UCVA) were 0. 01~0. 05, and the best spectacle-corrected visual acuity ( BSCVA) were 0. 4 ~ 1. 0. One month after surgery, the UCVA were 0. 5~1. 2. The mean vault were 547±222 μm (95%CI 442~672μm) and 528±268μm (95%CI 354 ~635μm) for 1mo and 1a, respectively (P = 0. 81), and there was no significant difference. Anterior subcapsular opacities in 1 eye, mild and transient increase in lOP in 3 eyes, and chronic pigment dispersion in 2 eyes were observed. There was no serious complication.
?CONCLUSlON: Posterior chamber phakic intraocular lens implantation is an effective and safe method for correcting patients with extreme highly myopia.

OBJECTIVETo observe the efficacy of He-wei Jie-gu (Chinese characters) decoction on femoral intertrochanteric fractures after internal fixation in elderly.

METHODSFrom January 2010 to January 2012, 90 elderly patients with osteoporotic intertrochanteric fracture were randomly divided into two groups. The patients in experimental group were treated with open reduction and internal fixation, He-wei Jie-gu decoction and Caltrate D in oral, including 18 males and 27 females with an average age of (77.8±11.3) years old ranging from 65 to 89 years old;the time from injured to operation was 2 to 9 days with an average of (6.6±2.1) days; according to Evans-Jensen classification, 17 cases were in type I, 8 in type II, 15 in type III, 3 in type IV, 2 in type V. Meanwhile, the patients in control group were treated with open reduction and internal fixation, Caltrate D in oral, including 16 males and 29 females with an average age of (81.2±9.6) years old ranging from 67 to 91 years old; the time from injury to operation was 2 to 10 days with an average of (6.8±2.6) days; according to Evans-Jensen classification, 15 cases were in type I, 10 in type II, 14 in type III, 4 in type W, 2 in type V. The VAS score, hip BMD value before and after treatment, fracture healing and the Harris function score after operation were recorded and compared.

RESULTSAll patients were followed up from 3 to 9 months with an average of (5.6±2.8) months. Compared with the control group, the VAS score of the experimental group had an obviously reduction (F=98.138, P<0.01) after treatment; the hip BMD value at 3 months after treatment in the experimental group was higher than that of the control group (P<0.05); the fracture healing rate of the experimental group was higher than that of the control group (P<0.05); Harris score in 3 months of follow-up in the experimental group was higher than that of the control group (t=2.542, P<0.05); all the differences between two groups were statistically significant (P<0.05).

CONCLUSIONHe-wei Jie-gu decoction can reduce postoperative pain and increase bone mineral density after internal fixation of femoral intertrochanteric fracture in elderly, promote the fracture healing and functional recovery in the hip joint.

Aged ; Aged, 80 and over ; Bone Density ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Femur ; injuries ; surgery ; Fracture Fixation, Internal ; Hip Fractures ; surgery ; Humans ; Male ; Osteoporotic Fractures ; drug therapy ; physiopathology ; surgery ; Pain, Postoperative ; drug therapy ; physiopathology

Objective Color doppler ultrasonography of chronic Keshan disease (CKD) was evaluated to provide evidences for clinic diagnosis of the disease. Methods From September to Novermber 2009, according to "Diagnostic criteria of Keshan disease" (GB 17021-1997), 64 cases of CKD were randomly sampled from five Keshan diseased districts in Shandong province, Zoucheng, Sishui, Yishui, Wulian, Jvxian, and Pingyi as patient group. Thirty four healthy volunteers being checked up by Shandong Institute for Endemic Diseases Control and Research were put in control group. All the subjects were examined with Color doppler ultrasonography. The indexes of cardiac structure, left ventricular (LV) systolic function and LV diastolic function were measured.Results Left atrial internal diameter, LV end-diastolic internal diameter, LV end-systolic internal diameter, right ventricular diameter, aorta diameter, right atrial transverse diameter, right atrial long diameter and left ventricle mass of the patient group[(35.38 ± 6.89), (61.57 ± 8.61), (45.39 ± 10.29), (17.22 ± 3.79), (28.69 ± 2.81),(38.00 ± 6.05), (42.68 ± 8.65)mm, (283.22 ± 103.12)g] were higher than that of control group[(26.70 ± 3.27),(45.41 ± 4.93), (26.91 ± 4.35), (13.76 ± 2.27), (24.09 ± 2.89), (31.50 ± 3.32), (35.82 ± 3.14) mm, (156.03 ±39.86)g, t = 6.93, 10.09, 9.98, 4.87, 7.64, 5.81, 4.46, 6.90, all P＜ 0.05]. The LV ejection fraction and fractional shortening of the left ventricular of the patient group[(49.25 ± 14.33)%, (26.11 ± 9.17)%] were lower than that of control group[(73.88 ± 4.04)%, (42.88 ± 3.62)%, t = - 9.79, - 10.22, all P＜ 0.05]. Diffuse hypokinetic motion of the left ventricle reduced in 95% (61/64) of CKD patients, and 5% (3/64) of CKD patients had segmental LV dyskinesia. Seventy five per sent(48/64) of the patients accompanied with mitral regurgitation, and 39% (26/64) of these cases accompanied with tricuspid regurgitation. Meaningful Mitral or tricuspid regurgitation was not found out in control group. Conclusions The CKD patients' bore of atrio-ventricular cavity and LV mass are enlarged, and their motion of ventricle is reduced or partly reduced. They have poor heart function. Mitral regurgitation are more than tricuspid regurgitation. Color doppler Ultrasonography is important in diagnosis of chronic Keshan discase.

Objective:To observe the efficacy of He wei Jie gu(和胃接骨)decoction on femoral intertrochanteric frac?tures after internal fixation in elderly. Methods:From January 2010 to January 2012,90 elderly patients with osteoporotic in?tertrochanteric fracture were randomly divided into two groups. The patients in experimental group were treated with open re?duction and internal fixation,He wei Jie gu decoction and Caltrate D in oral,including 18 males and 27 females with an aver?age age of(77.8±11.3)years old ranging from 65 to 89 years old;the time from injured to operation was 2 to 9 days with an av?erage of(6.6±2.1)days;according to Evans-Jensen classification,17 cases were in typeⅠ,8 in typeⅡ,15 in typeⅢ,3 in typeⅣ,2 in type V. Meanwhile,the patients in control group were treated with open reduction and internal fixation,Caltrate D in oral,including 16 males and 29 females with an average age of(81.2±9.6)years old ranging from 67 to 91 years old;the time from injury to operation was 2 to 10 days with an average of(6.8±2.6)days;according to Evans-Jensen classification,15 cases were in typeⅠ,10 in typeⅡ,14 in typeⅢ,4 in typeⅣ,2 in typeⅤ. The VAS score,hip BMD value before and after treatment,fracture healing and the Harris function score after operation were recorded and compared. Results:All patients were followed up from 3 to 9 months with an average of(5.6±2.8)months. Compared with the control group,the VAS score of the experimental group had an obviously reduction(F=98.138,P<0.01)after treatment;the hip BMD value at 3 months after treat?ment in the experimental group was higher than that of the control group(P<0.05);the fracture healing rate of the experimental group was higher than that of the control group(P<0.05);Harris score in 3 months of follow up in the experimental group was higher than that of the control group(t=2.542,P<0.05);all the differences between two groups were statistically significant(P<0.05). Conclusion:He wei Jie gu decoction can reduce postoperative pain and increase bone mineral density after internal fixa?tion of femoral intertrochanteric fracture in elderly,promote the fracture healing and functional recovery in the hip joint.

Objective To determine the prevalence and distribution of type 2 diabetes mellitus (T2DM) and the relationship between maximum body mass index (MAXBMI) and T2DM. Methods From June to August, 2005, a stratified cluster sampling of 1071 permanent residents in communities, over 20 years old, from 4 districts and 1 county of Mudanjiang was chosen. The prevalence of T2DM, and the association between T2DM and different levels of the MAXBMI, current BMI were studied. Results The prevalence in the communities was 7.09% and in those with past maximum BMI≥28 kg/m~2, it was 12.10%. With the increase of past MAXBMI levels, the risk of T2DM patients also increased significantly(trend X~2=17.387 23, P<0.0001). Data from multifactor analysis showed that MAXBMI in the past was positively related to T2DM (OR=3.06, P=0.0013). In T2DM patients, the group with MAXBMI≥27.4 kg/m~2 had higher 2-hour postprandial blood glucose than those with lower MAXBMI (P=0.0408). When compared with low maximum BMI group in normal blood glucose population, the group with higher MAXBMI (≥ 25.4 kg/m~2) had higher blood glucose and greater change of BMI. Conclusion In both groups that patients with T2DM and with normal glucose, in order to control blood glucose better, researchers should not only concern about the influence of the MAXBMI in the past, but also pay attention to constantly keep BMI at the normal range.

Syncope is a common emergency of children and adolescents,which has serious influence on the quality of life.Neurally-mediated syncope,including postural tachycardia syndrome,vasovagal syncope,orthostatic hypotension and orthostatic hypertension,is the main cause of syncope in children and adolescents.The main manifestations of neurally-mediated syncope are diverse,such as dizziness,headache,chest tightness,chest pain,pale complexion,fatigue,pre-syncope and syncope.Although the clinical manifestations are similar,each subtype of syncope has its hemodynamic feature and optimal treatment option.The diagnosis rate of syncope in children has been greatly improved on account of the development of the diagnostic procedures and methods.In recent years,with the promotion of head-up tilt test and drug-provocated head-up tilt test,the hemodynamic classification of neurally-mediated syncope gets continually refined.In recent years,with the effort of clinicians,an appropriate diagnostic protocol for children with syncope has been established.The initial evaluation consists of history taking,physical examination,standing test and standard electrocardiography.After the initial evaluation,some patients could be diagnosed definitely,such as postural tachycardia syndrome,orthostatic hypotension,and situational syncope.Those with a specific entity causing syncope need selective clinical and laboratory investigations.Patients for whom the cause of syncope remained undetermined should undergo head-up tilt test.The precise pathogenesis of neurally-mediated syncope is not entirely clear.In recent years,studies have shown that neurally-mediated syncope may be related to several factors,including hypovolemia,high catecholamine status,abnormal local vascular tension,decreased skeletal muscle pump activity and abnormal neurohumoral factors.Currently based on the possible pathogenesis,the individualized treatment of neurally-mediated syncope has also been studied in-depth.Generally,the management of neurallymediated syncope includes non-pharmacological and pharmacological interventions.Patient education is the fundamental part above all.In addition to exercise training,the first-line treatments mainly include oral rehydration salts,beta adrenoreceptor blockers,and alpha adrenoreceptor agonists.By analyzing the patient's physiological indexes and biomarkers before treatment,the efficacy of medication could be well predicted.The individualized treatment will become the main direction in the future researches.

Objective To observe the influence of long-term treatment with inhaled corticosteroid on biochemical bone indexes and bone mineral density (BMD) in children with asthma. Methods The design was a randomized, paralleled group study with 3 low dose regiments of 100, 200,300 micrograms of budesonide per day in 45 children with asthma aged 5-8 years old for 12 months. Before inhaled corticosteroid therapy and 6th,12th month,clinical effects were observed and lung function(FEV1) was measured; concentration of serum osteocalcin(OST),insulin-like growth factor-1(IGF-1),bony alkaline phosphatase (BALP) and urinary deoxypyridinolin: creatinine (DPD/Cr) were measured; BMD was examined by dual energy X-ray absorptiometry. Results Clinical evaluation was improved and there was significant increase in FEV1 of asthmatic children. The amount of serum OST was slightly higher,yet no significant compared with that of normal control group. There was significant increase of serum BALP in asthmatic children after treatment; there was significant increase in serum IGF-1 of patients group after treatment compared with in normal children at the same age group; there was significant decrease in urinary DPD/Cr after treatment.There was no significant decrease in BMD before and after treatment at the hip (neck of femur , trochanter of femur ,Ward′s triangle),the lumber area of the spine (L2-4) and forearm (ultradis, distal). Conclusion Long-term treatment with low does corticosteroid dose not restrictedly affect bone metabolism and BMD in children with asthma.

OBJECTIVETo analyze the genotype and phenotype correlation in the hemoglobin E (HbE) carriers, and to investigate the effect of HbE on hematological parameters.

METHODSThe capillary electrophoresis was used to screen total 14 141 samples and blood cell analysis was further processed to the HbE carrying samples. Gap-PCR and reverse dot blot hybridization method were used for the detection of Chinese common mutation of α and β thalassemia.

RESULTSThere is a statistical difference in hematological phenotype index (HGB, MCV, MCH, HbE, HbA(2)) between samples of HbE heterozygous (53 samples), HbE homozygous (2 samples), HbE composite α thalassemia (α-thal, 7 samples) and HbE composite β thalassemia (β-thal, 8 samples). Among the four groups, HbE heterozygous \[HGB (122.7 ± 19.99) g/L, MCV (78.65 ± 5.03) fl\] and HbE composite α-thal \[HGB (113.6 ± 22.68) g/L, MCV (73.50 ± 7.73) fl\] had slight effect on hematological parameters, but HbE composite β-thal \[HGB (76.4 ± 12.30) g/L\], MCV (59.23 ± 5.28) fl\] had the heaviest effect on hematological parameters.

CONCLUSIONCo-existence of HbE heterozygous and other type thalassemias showed variation in their hematological phenotype, so patients should be informed of genetics in prenatal diagnosis.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Genotype ; Hemoglobin E ; genetics ; Heterozygote ; Humans ; Infant ; Male ; Middle Aged ; Phenotype ; Thalassemia ; blood ; genetics ; Young Adult

OBJECTIVETo explore the role of X-linked inhibitor of apoptosis protein (XIAP) in the fibronectin (Fn)-adhesion mediated drug resistance of HL-60 cells.

METHODSCulture plates were coated with Fn and bovine serum albumin (BSA) (as control), respectively. Colorimetric CCK-8 assay was used to determine the effects of Fn on the cytotoxicity of DNR to HL-60 cells. Intracellular DNR accumulation was assayed with flow cytometry. Reverse transcription-PCR and Western blot were used to examine the mRNA expression and XIAP, bcl-2, MRP and mdr1 proteins, respectively. HL-60 cells were added to Fn coated Culture plates. The fully phosphorothioate antisense oligonucleotide (AS-ODNs) and the control ODNs of XIAP were delivered into HL-60 cells in the form of liposome-ODN complexes. IC(50) was calculated by linear regression of survival percent versus drug concentration.

RESULTSHL-60 cells adhered to Fn-coated plates had a significant survival advantage over those grown on BSA coated plates and in suspension when exposed to DNR, the IC(50) of Fn group being significantly higher than that of BSA group and suspension group (0.526 micromol/L vs 0.132 micromol/L, 0.123 micromol/L, respectively, P < 0.05). XIAP was up-regulated significantly in Fn group compared with BSA group and suspension group (P < 0.05), whereas there was no difference in the expressions of bcl-2, MRP and mdr1 among the three groups (P > 0.05). The intracellular concentration of DNR in Fn-adhered HL-60 cells was similar to that in BSA group and suspension group (P < 0.05). AS-ODNs of XIAP down-regulated the XIAP expression in Fn-adhered HL-60 cells. In addition, AS-ODNs sensitized HL-60 cells to the cytotoxic effects of DNR.

CONCLUSIONThe increased XIAP protein level contributes to the drug resistance induced by adhesion to Fn. AS-ODNs of XIAP might reverse the drug resistance.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Cell Adhesion ; Daunorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; physiology ; Drug Resistance, Neoplasm ; genetics ; physiology ; Fibronectins ; HL-60 Cells ; Humans ; Multidrug Resistance-Associated Proteins ; genetics ; metabolism ; Oligonucleotides, Antisense ; genetics ; RNA, Messenger ; genetics ; Transfection ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism ; physiology