Objective To study the effective methods for extraction of clenbuterol from biological material. Methods Clenbuterol in blood samples was extracted with 5 different methods,the extracts from various extraction methods were determined by gas chromatography-mass spectrometry(GC-MS?雪. The efficiency of 5 extraction methods was compared with each other. Results The standard curve showed good linear relationship with a range from 0.01-10.0 mg/L clenbuterol hydrochloride(r=0.999 9?雪. The detection limit was 0.005 mg/L. After adding 1.0 ?g/ml clenbuterol hydrochloride into blood sample,the blood sample was immersed and extracted in 0.01 mol/L HCl and absolute alcohol.The extracts were extracted by chloroform at pH value of 11. The average recovery and RSD of this method were 96.5% and 5.21% respectively(n=6?雪. Conclusion The method showed easy operation and reliable results ,it was suitable for determination of clenbuterol in biological material without deviation.

Objective To determine whether tissue factor(TF) is present in the amniotic fluid of normal term pregnancy. Methods Tissue factor antigen levels in amniotic fluid, blood plasma, centrifugal supernatant and sediment of 16 normal term pregnancies were detected with immunoenzymatic method(ELISA). Results Tissue factor antigen levels in the blood plasma, the whole amniotic fluid, the centrifugal supernatant and the sediment were [(31?9) ng/L0, [(404?186) ng/L], [(348?177) ng/L] and [(1360?639) ng/L], respectively. Tissue factor antigen concentrations in three states of the amniotic sediment(P

OBJECTIVE To study the cytotoxicity of polyamide amine dendrimers (PAMAM)-hyaluronic acid(HA)as a non-viral gene delivery vector in vitro. METHODS PAMAM-HA was synthesized by our laboratory. Cytotoxicities of various polymers on HeLa cells,Bel-7402 cells and HepG2 cells were evaluated by MTT assay. Apoptotic rates were determined by flow cytometry. RESULTS The cytotoxicity of PAMAM-HA polymer increased with the concentration of polymer(50-800 mg · L-1),the time of action(24-72 h), the number of generations of PAMAM(G4,G5) and decrease in the molecular mass(3850,17 200)and the graft density(5%-25%)of HA. After incubation with PAMAM G4-HA3850-5%or PAMAM G5-HA3850-5%for 24 h,the apoptotic rate of hepatoma cells—Bel-7402 cells was 4.5%and 9.9%,respectively. After incubation with complexes of PAMAM G4-HA3850-5%/DNA or PAMAM G5-HA3850-5%/DNA for 24 h,the viability of HeLa cells was more than 80%,which was lower than that of PAMAM G4/DNA和PAMAM G5/DNA. CONCLUSION Cytotoxicity of PAMAM modified by HA of different grafting density and quantity can be reduced ,suggesting the PAMAM is a promising non-viral gene vector.

Objective To explore the impacts of catechol-O-Methyltransferase (COMT) gene polymorphisms on the outcomes of methylphenidate treatment for attention deficit hyperactivity disorder (ADHD) in children. Methods One hundred seventy-seven ADHD children of Chinese Han descent received open-labelled dose titration with methylpheni?date to achieve optimal response in 2~4 weeks. The behavior changes were evaluated by using ADHD diagnostic scale (parent version) before and after treatment. COMT gene rs4680 (Val158Met) and rs165599 were genotyped using fluores?cent real-time PCR. The genotype distribution and treatment outcomes including remission, response and non-response were analyzed. Results The treatment response differed significantly among patients with different genotypes of rs4680 (P<0.01). Thirty-eight (40.9%) patients with G/G genotype had remission and 19 (20.4%) had good response;Thirty-five (48.6%) patients with G/A genotype had remission and 16 (22.2%) had good response. In contrast, only one patients with A/A genotype had remission and none of them had good response. The changes of the ADHD diagnostic scale total scores, inattention scores and hyperactivity impulsive scores were statistically different among different genotypes of rs4680 (P<0.05). The hyperactivity impulsive scores were significantly higher in the G allele carrier than A allele carrier (P=0.04). The treatment response was not significantly different among different rs165599 genotypes (P>0.05). Conclu?sion COMT gene rs4680 (Val158Met) polymorphism is associated with methylphenidate response in a Han Chinese popu?lation. Patients with G allele is more likely to benefit from methylphenidate treatment in comparison with A/A genotype.

Objective: To evaluate the clinical efficacy and side effects of icotinib combined with a subarachnoid space implantable pump for the treatment of leptomeningeal metastases (LM) from lung adenocarcinoma. Methods:Seven cases of LM from lung adeno-carcinoma with epidermal growth factor receptor mutation were included in the study from March 2011 to September 2015. With the aid of anesthetists, all patients were implanted with subarachnoid space implantable pumps to drain the cerebrospinal fluid (CSF) and then treated with icotinib (125 mg, three times a day) until disease progression or unacceptable toxicities. After 4 weeks, the efficacy and tolerability of icotinib were evaluated on the basis of symptoms, tumor markers from CSF, and brain gadolinium-enhanced magnet-ic resonance imaging scans. Results:Among the seven patients evaluated, no patient had complete response, two patients had partial response, four had stable disease, and one had progression. The patient who was progressive died at a month after therapy. The sur-vival time of all the other patients was more than 4 months. The common adverse effects of icotinib were skin rash and diarrhea, main-ly in grades 1 and 2. No infection of the local subarachnoid drainage tube was found in the abdominal wall. Conclusion:Icotinib com-bined with a subarachnoid space implantable pump for the treatment of LM from lung adenocarcinoma may be effective, and the tox-icities are tolerable. This method could also obviously alleviate the clinical symptoms and improve the quality of life of the patients, worthy of further study.

Atherosclerosis (AS) is a common underlying disease that hazard to human health.It is closely associated with stroke,hypertension,coronary artery disease,and diabetes.AS on the premise of vascular intimnal injury,excessive smooth muscle cell proliferation,inflammatory cell infiltration,oxidative stress,apoptosis and other mschanisms participate together.In eucaryotic organisms,most proteins are degraded by ubiquitin-proteasome system (UPS).An increasing number of evidence indicates that UPS is associated with the occurrenee and development of AS and complications.This article reviews the relationship between.UPS and AS,as well as the effect of ubiquitin-proteasome inhibitor on AS.

Objective To explore the effect of dexmedetomidine on hemodynamics in hypertensive patients undergoing thyroid surgery with local anesthesia.Methods Sixty patients with preoperatively diagnosed class Ⅰ to Ⅱ hypertension undergoing selective thyroidectomy were randomly divided into D (dexmedetomidine) and M (midazolam) groups (30 patients in each group).Doses of dexmedetomidine 0.5 μg/kg were finished in 20 minutes injection in patients of D group before the start of the surgery,then sequentially maintained at the rate of 0.1 ～ 0.5 μg/ (kg · h) with decreasing speed of 0.1 μg/(kg · h) when systolic pressure kept down to 110 mmHg or heart rare down to 60 bpm or Ramsay score of 3 points.The patients in M group were injected with midazolam 0.04 mg/kg 20 minutes before the start of surgery,then maintained at the rate of 0.02 ～ 0.05 mg/(kg · h),decreasing speed of 0.1 mg/(kg · h) to keep Ramsay score of 3 points if necessary.Two groups of patients with Ramsay score of 3 points but a high blood pressure (higher than 30％ of basic level) or heart rate (more than 100 bpm) were treated with drugs during operation.Record the systolic pressure,diastolic pressure,heart rate and Ramsay score at the time of before medication (T0),the injection of local anesthetics (T1),the start of surgery (T2),pain compliments required additional local anesthetics (T3),dealing with gland (T4) and the end of surgery,VAS in 8 h after operation and adverse reaction were recorded.Results In D group,the dosages of urapidil and esmolol [(10 ±5)mg and (0 ±0)mg] were significantly less than those in patients of M group [(60 ± 10) mg and (80 ±5) mg,t =14.82,t =19.78,P ＜ 0.05].Blood pressure and heart rate were all significantly decreased at the time of T1,T2 and T5 when comparing with T0(P ＜0.05 orP ＜0.01),and only heart rate was significantly decreased at the time of T3 and T4 (P ＜ 0.05).While in M group,blood pressure and heart rate were higher than basic levels at the time of T3 and T4 (P ＜ 0.05).Besides,lower blood pressure and heart rate were less than those in M group at all observed time expect T0 (P ＜ 0.05).Conclusions Good sedation effects can be produced by both dexmedetomidine and midazolam in hypertensive patients undergoing thyroid surgery with local anesthesia,but dexmedetomidine was determined more suitable in sedation and anti-hypertension in patients with light to moderate hypertension for better hemodynamic stability effect with local anesthesia.

Objective To investigate the effects of oestrogen-receptor(ER), progesterone-receptor(PR), vascular endothelial growth factor(VEGF) and kinase insert domaincontaining receptor(KDR) in uterine bleeding induced by intrauterine device(IUD). Methods Immunohistochemical method was used to detect the expressions of ER, PR, VEGF and KDR in endometrium from IUD bleeding group and normal control group, respectively. Microvessel density(MVD) was evaluated by using anti factor Ⅷ-related antigen (F8-RA) antibody to mark stromal microvessels. Results The expressions of ER, VEGF and KDR were significantly higher in IUD-induced endometrium bleeding group than those in control group(P0.05). MVD was significantly higher in IUD-induced endometrium bleeding than that in normal endometrium(P

Objective To study the associations between neo-adjuvant chemotherapy (NACT)sensitivity and expression of relative proliferation and apoptosis genes in cervical cancer tissues. Furthermore,the potential roles of relative genes expression as monitor in the chemotherapy against cervical cancer tissues were studied. Methods Fifteen pathologically proved Ⅱ A stage cervical cancer patients with HPV 16 positive were divided into effective group and non-effective group according to the clinical response to NACT, the changes of HPV16-E6, p53 genes and proteins expressions were measured by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot. At the same time, MTT assay was used to detected proliferative activity to paclitaxel liposome and carboplatin (paraplatin) in HeLa cells. Results In effective group, the overall response rate was 73.3 % (11/15) and the HPV16-E6 mRNA and protein level after NACT was significantly decreased than that before NACT (t=3.359, t=3.614, P＜0.05), while p53mRNA and protein level increased significantly (t =5.852, t=2.838, P ＜0.05). However, there have no significant different between pre-NACT and Post-NACT of the two genes in the non-effective group. After using chemotherapy HeLa cells growth decreased. Conclusion HPV16-E6 and p53 expreesion level are useful paraneters in evaluating NACT response in cervical cancer tissues. The detection of the two genes expression is a method for predicting efficacy in NACT.

Objective:To investigate the association between LPHN3 andattention-deficit/hyperactivity disor-der(ADHD)in Chinese Han children.Methods:Based on the Diagnostic and Statistical Manual of Mental Disor-ders,Fourth Edition (DSM-IV)diagnosis criteria,921 normal controls and 1052 ADHD children were included in the study.The Clinical Diagnostic Interview Scale (CDIS )was used to assess symptoms and ADHD sub-types.ADHD was divided into three subtypes,namely ADHD inattentive type (ADHD-I),ADHD hyperactive-im-pulsive type (ADHD-HI),and ADHD combined type (ADHD-C).The ADHD rating-scale was used to assess ADHD symptoms.Including inattentive symptoms,hyperactive symptoms,impulsive symptoms and ADHD total symptoms.Three single nucleotide polymorphisms (SNPs)of LPHN3 were genotyped.Case-control studies were conducted to investigate the association of each SNP with the ADHD and the subgroups using chi-square test.Results:rs11131347 was associated with ADHD P <0.05,OR =0.86(0.76 -0.98)],but the difference didn't survive significance after corrections,The frequency of minor allele C in cases and control was0.409 vs.0.445.For different genders,rs11131347 was significantly associated with ADHD boys [P <0.05,OR =0.82(0.71 -0.96)], The frequency of minor allele C in cases and control was0.402 vs.0.449.For different subtypes,rs11131347 was associated with ADHD-C [P <0.05,OR =0.85(0.74 -0.98)]and ADHD-C boys[P <0.05,OR =0.82(0.70 -0.97)],but none of the difference survived significance after corrections,The frequencies of minor allele C in cases and control were respectively 0.407 vs.0.445 and 0.401 vs.0.449.Genotypes distribution analysis indicated that rs11131347 was associated with ADHD in general (recessive model,P <0.05),ADHD boys (additive model,P <0.05;dominant model,P <0.05;recessive model,P <0.05),ADHD-C(recessive model,P <0.05)and ADHD-C boys (dominant model,P <0.05 ),however,none of the difference survived significance after correc-tions.rs11131347 was nominal associated with impulsive scores(P <0.05).Conclusion:These findings suggest that the polymorphism of LPHN3 is probably involved in the pathological mechanisms of ADHD and its core symptoms of impulsivity.