Objective To construct the recombinant plasmid that highly expressed human subcellular PTEN,in order to provide a basis for further study on its anti-tumor effect. Methods PTEN cDNA was amplified by RT-PCR based on mRNA of placenta.The PCR product was ligated into T-vector,and transfected into E.coli;the obtained T-PTEN plasmid was identified with restrictive digestion and sequencing.PCR was used to incorporte nuclear signal of localization(NSL) into PTEN when T-PTEN was used as template.Then the PCR product was ligated into T-vector,and transfected into E.coli,and T-NSL-PTEN plasmid was obtained.pcDNA3.1 and T-NSL-PTEN were ligated after digested with EcoRⅠand BamHⅠ,and transfected into E.coli,the recombinant vector pcDNA3.1-NSL-PTEN was obtained,and identified with digestion and sequcncing.Results The recombinant expression vector DUM-PTEN and PUM-NSL PTEN were identified by restrictive digestion and DNA sequencing.As expected,by EcoRⅠ and BamHⅠ digestion,it showed the band of 1 200 bp.The sequencing result showed the NSL was incorporated successfully.The recombinant pcDNA3.1-PTEN was obtained with 1 200 bp,the sequencing result showed that its sequence was same as target gene;the recombinant pcDNA3.1-NSL-PTEN was comfirmed by restrictive digestion and sequencing,and the NSL was incorporated successfully. Conclusion The recombinant expression plasmid pcDNA3.1-NSL-PTEN is constructed successfully which can highly express human subcellular PTEN.

Objective To explore the clinical application value of serum amyloid protein(SAA) and C reactive protein(CRP)in early diagnosis of hand‐foot‐mouth disease(HFMD)in children .Methods The serum/plasma levels of CRP and SAA in 81 children with HFMD(HFMD group)and 125 healthy children(healthy control group)were detected by using dot immunogold filtration as‐say .Data including white blood cell (WBC) count ,levels of CRP and SAA were obtained and analyzed .And diagnostic efficacies of these indicators were evaluated by using receiver operating characteristic (ROC) curve .Results The serum levels of SAA and CRP and WBC count in HFMD group were significantly higher than those in healthy control group ,and all had statistically significant differences(P<0 .05) .The area under the ROC curve(AUC) of SAA(0 .921) was higher than that of CRP(0 .728) ,SAA/CRP ra‐tio(0 .846) and WBC count(0 .721) for diagnosing HFMD ,there were statistically significant differences(P<0 .05) .Compared with CRP ,WBC count and SAA/CRP ratio ,SAA had better diagnostic efficacy .Conclusion As a new biomarker of infections ,SAA can effectively diagnose HFMD early and provide more sensitive reference to clinicians for early diagnosis ,which might be worthy of clinical application widely .

Tibetan medicineLiu-Wei Mu-Xiang(LWMX) pill, a traditional compound medicine, has special therapeutic effects in the treatment of gastrointestinal anabrosis, especially chronic gastric ulcer. It has been widely applied in the clinics in Tibetan region. Nevertheless, the current standards for quality control are insufficient and incomplete, which makes it difficult to ensure its quality. This review focused on two closely related perspectives, which were the effective material basis and quality control, to summarize and review the recent advancement in relevant studies. This review also pointed out that the burning questions for Tibetan medicine standards were the origin confusion and“different medicines with the same name”. It offered meaningful suggestions for the consideration to improve its quality standards.

An ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF-MS)-based chemical analytic technology was used to evaluate the chemical constitution of Radix Aconiti Lateralis Preparata in the process of decocting, so as to provide a scientific basis for processing Radix Aconiti Lateralis Preparata.

To evaluate whether Shenfu injection (SFI) protects against cardiac myocyte injury induced by Fupian injection (FPI) in vitro.

OBJECTIVE Tostudythehepatotoxicityofveratrinehydrochloride(VH)anditsmecha-nismoninductionofapoptosisinvitro.METHODS HepG2cellswereexposedtoVH0.1-0.6g·L-1 for 24 h,cell viability was examined by CCK-8 assay,and the morphologic changes in HepG2 cells were quantified.After the treatment with VH 0.1 -0.5 g·L-1 for 24 h,cell membrane injury was examined by detecting the release rate of lactate dehydrogenase (LDH).The effect on reactive oxygen species (ROS),mitochondrial membrane potential and apoptosis was detected by flow cytometry.The mRNA expression of p53,Bax,cytochrome c,caspase 9,caspase 3 was evaluated by real-time PCR. RESULTS HepG2cellviabilitywassignificantlyreducedfollowingexposuretoVH0.1-0.5g·L-1. The IC50 value was 0.4 g·L-1 .The 95%confidence limit was 0.2558-0.6965 g·L-1 .The LDH release rate,ROS and apoptosis rate of HepG2 cells were significantly increased after exposure to VH 0.1 -0.5 g·L-1 for 24 h (P<0.05,P<0.01 ),and the mitochondrial membrane potential markedly declined (P<0.05,P<0.01 ).The expression of p53,Bax,cytochrome c,caspase 9 and caspase 3 was increased(P<0.05,P<0.01).CONCLUSION VHhascytotoxicpotential.Damagetocell me mbrane and mitochondria and initiation of apoptosis-related genes of caspase 9 and caspase 3 mRNA expression may be the mechanis m of apoptosis.

OBJECTIVE To invesigate the prevalence of SHV-producing Enterobacteriaceae isolates in Hunan Province and identify the subtype of SHV encoding gene and the epidemiological aspect of SHV-producing isolates. METHODS Isolates were collected and identified as well as subjected to ESBLs detection.PCR and DNA sequencing were used to determine the genotype of SHV enzymes.The homology of SHV-producing strains were detected by RAPD. RESULTS Twenty-six of 171 Enterobacteriaceae isolates were confirmed to produce blaSHV genes,and 5 subtypes of SHV-type ?-lactamases were determined,including 9 strains of SHV-28,7 strains of SHV-12,7 strains of SHV-1,2 strains of SHV-11 and 1 strain of SHV-5.There were 6 RAPD types in 19 isolates of SHV-producing Klebsiella pneumoniae,5 RAPD types in 5 isolates of SHV-producing Enterobacter cloacae. CONCLUSIONS SHV-12 is the predominant genotype of SHV ESBLs producing Enterobacteriaceae in Hunan Province,and clone spread has played a certain role in SHV-type ?-lactamase producing K.pneumoniae.SHV ESBLs are not found in Escherichia coli.

Objective To study the prognostic and predictive significance of 21-gene assay ( oncotype DX)in breast cancer.Methods Real-time quantitative PCR(RT-QPCR)was used to detect 21 gene expression in breast cancer tissues (100 cases)and recurrence score(RS)was calculated.Results Among the 100 cases, 52 cases had low RS , 22 cases had middle RS , and 26 cases had high RS .The recurrence rate of five years was 1.92%,4.55%and 15.38%respectively.21 gene expression had nothing to do with patients'age, tumor size, histological grade , lymph node metastasis state , ER expression , or PR expression .It was associated with HER 2 expression .Conclusions 21 genes is a good prediction factor in breast cancer and its prognosis .

Objective To discuss the significance of serum IgE before and after inhale glucocorticoid treat-ment of children's asthma. Methods 520 children with asthma were seleceted from the outpatient. Different type of fluticasone propionate were given to different age groups: Aerosol type by a spacer in less than 5 years old,and in-halant (Seretide) 5 years and the above. The dosage was between 200 μg/day to 375 μg/day. IgE was tested before and 3 months after the treatment. Results Serum IgE decreased significantly in 3 months treatment [ from (496.12±24.75) kU/L to (390.71±18.71) kU/L] (t=7.337,P<0.01). The change of IgE was related to clinical effect and age. The level increased in those less than 3 years [(307.05±34.71)kU/L vs (483.09±41.78) kU/L] (t=2.963,P=0.004),but decreased between 4 to 5 years old group [(543.46±51.03) kU/L vs (316.93±29.30) kU/L] (t=3.368,P=0.000) ,and decreased between 6 to 14 years old group[ (586.30±37.19)kU/L vs (387.61±27.60) kU/L] (t=4.827,P=0.000). In fluticasone group IgE level changed from (468.91±32.81) kU/L to (359.03±22.79) kU/L after treatment (t=5.988,P<0.01),which decreased from (586.30±37.19) kU/L to (387.6±27.60) kU/L in Salmeterol group (t=4.827,P<0.01). In 260 cases of IgE below 300 kU/L 109 cases (41.92%,109/260) increased while in 260 cases of IgE above 300 kU/L,total IgE lev-el increased in 45 cases (16.15% ,45/260) after treatment,with statistical significance(χ<'2>=37.789,P=0.000). Conclusion Inhale glucocorticoid can make the level of IgE decreased.

In the present study, an ultra performance liquid chromatography coupled with time-of-fight mass spectrometry (UPLC-TOF/MS) based chemical profiling approach was used to evaluate chemical constitution between co-decoction and mixed decoction of ginseng and Radix Aconiti Praeparata. Two different kinds of decoctions, namely co-decoction of ginseng and Radix Aconiti Praeparata: water extract of mixed two herbs, and mixed decoction of ginseng and Radix Aconiti Praeparata: mixed water extract of each individual herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-TOF/MS analysis. The datasets of t(R) m/z pairs, ion intensities and sample codes were processed with supervised partial least squared discriminant analysis (OPLS-DA) to holistically compare the difference between these two decoction samples. Significant difference between the two decoction samples was showed in the results of positive ion mode. The contents of hypaconitine and deoxyaconitine decreased, while that of benzoylmesaconine, benzoylhypaconine and dehydrated benzoylmesaconine increased in the samples of co-decoction of ginseng and Radix Aconiti Praeparata. The content of diester-diterpenoid alkaloids decreased, while that of monoester-diterpenoid alkaloids increased, which is probably the basis of toxicity-attenuated action when combined ginseng with Radix Aconiti Praeparata.