A strain Escherichia coli with high production of phytase was screened from pig excreta. Phytase gene appA, with 1,299 bp coding region in full length, was cloned from its genome by polymerase chain reaction (PCR) . The gene appA was then cloned into the prokaryotic expression vector pET-28a ( + ) . In the host BL21, the phytase appA was overexpressed by shaker-cultivation (up to 692 U/mL) . The enzymatic analysis of the prokaryotic derived appA phytase revealed that its optimal pH and temperature was 4.5 and 60℃, respectively.

BACKGROUNDOver-expression of epidermal growth factor receptor (EGFR) is thought to be related to cell proliferation, invasion, metastasis, resistance to chemoradiotherapy and poor prognosis of various human cancers. Forty percent to fifty percent of glioblastoma multiforme (GBM) possess deregulated EGFR, which may contribute to the aggressive and refractory course of GBM. Therefore, blockade of EGFR signal transduction may be a promising treatment strategy for GBM.

METHODSMTT assay, cell growth curve assay and tumor xenograft model were used to evaluate the antitumor activity of F90 against SHG-44 in vitro and in vivo. Western blot assay was applied to evaluate the expression of p-EGFR, p-ERK1, p-JNK, p-P38, Bcl2 and P53 proteins.

RESULTSF90 inhibited the cell proliferation in a dose-dependent manner in vitro. The growth of SHG-44 tumor xenografts was suppressed by F90 at a high dose level (100 mg x kg(-1) x d(-1)). Phosphorylation of EGFR and activated downstream signaling proteins, such as ERK1, JNK and P38, were found to be depressed after incubation with F90 for 48 hours in vitro. Down-regulated Bcl2 protein and up-regulated P53 protein were also observed.

CONCLUSIONSThe results demonstrate that F90 is effective in inhibiting the proliferation of SHG-44 cells in vitro and tumor growth in vivo, suggesting that F90 may be a new therapeutic option for treatment of GBM.

Animals ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Glioblastoma ; drug therapy ; pathology ; Humans ; MAP Kinase Signaling System ; drug effects ; Mice ; Mice, Inbred BALB C ; Phosphorylation ; Protein Kinase Inhibitors ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Quinazolines ; pharmacology ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; metabolism ; Tumor Suppressor Protein p53 ; analysis

OBJECTIVE To explore the neuro-protective effects of saffron (Crocus satius L.) on chronic focal cerebral ischemia in rats.METHODS SD rats were randomly divided into 6 groups:sham control group,MCAO group,edaravone group and saffron 30,100,300 mg·kg-1groups.Focal cerebral ischemia was induced by middle cerebral artery occlusion(MCAO).Saffron was administered orally by once daily from 2 h to 42 d after ischemia. At 42 d after cerebral ischemia, neurological deficit score, spontaneous activity test,elevated plus maze test,marble burying test and novel objective recognition test were used to evaluate the effects of saffron on the behevioural change. Infarct volume, survival neuron density, activated astrocyte number, and the thickness of glial scar were also detected. GFAP expression and inflammatory cytokine contents in ischemic peripheral region were detected by Western blot and ELISA,separately.RESULTS Saffron(100,300 mg·kg-1)improved the body weight decrease, neurological deficit and spontaneous activity. Saffron (30-300 mg·kg- 1) increased the traveled distance ratio and total time in open arm, decreased the buried marble number, which indicated that saffron could ameliorate anxiety- and depression-like behaviors. Saffron (100, 300 mg·kg-1)improved the learning and memory function,which manifested by increased discrimination ratio(DR)and discrim-ination index (DI) in T2test. The results of toluidine blue found saffron treatment (100, 300 mg·kg-1) decreased the infarct volume and increased the neuron density in cortex and hippocampal.The activated astrocyte number,the thickness of glial scar and GFAP expression in ischemic peripheral region decreased after saffron. Saffron (100, 300 mg·kg-1) decreased the contents of IL-6 and IL-1β, increased the content of IL-10 in ischemic peripheral region.CONCLUSION Saffron exerted neuro-protective effects on chronic focal cerebral ischemia,which could be related with inhibiting the activation of astrocyte and glial scar,following with the decrease of inflammatory reaction.

Objective:To investigate the effects of chest pain center construction in basic-level hospitals on treatment time and short-term prognosis in patients with acute ST-elevation myocardial infarction.Methods:A total of 162 patients with acute ST-elevation myocardial infarction who received percutaneous coronary intervention (PCI) in The First People's Hospital of Jiande between November 2014 and November 2018 were included in this study. Among them, 66 patients who received treatment in The First People's Hospital of Jiande between November 2014 and October 2016 were included in the control group. The remaining 96 patients who received treatment between November 2016 and November 2018 were included in the study group. The underlying diseases, PCI success rate, first medical contact-to-balloon time, door-to-balloon time, in-hospital mortality, incidence of heart failure on the next day of PCI, length of hospital stay, hospital medical cost were retrospectively analyzed.Results:There were no significant differences in underlying disease composition ratio and PCI success rate between the two groups (both P > 0.05). There were significant differences in first medical contact-to-balloon time [(185.2 ± 53.7) minutes vs. (108.6 ± 46.4) minutes, t = 6.128], door-to-balloon time [(121.5 ± 23.2) minutes vs. (68.7 ± 14.3) minutes, t = 7.341], length of hospital stay [(10.3 ± 3.5) days vs. (7.2 ± 2.8) days, t = 5.128], hospital medical cost [(43 582.0 ± 7 186.5) yuan vs. (35 479.0 ± 4 213.1) yuan, t = 8.361], in-hospital mortality [6.1% vs. 3.1%, χ2 = 4.784], the incidence of heart failure on the next day of PCI [13.6% vs. 4.2%, χ2 = 8.253] between the control and study groups (all P < 0.05). Conclusion:Establishment of a standardized chest pain center construction in basic-level hospital can greatly shorten the first medical contact-to-balloon time, door-to-balloon time and length of hospital stay, improve the cardiac function and prognosis of patients with myocardial infarction, and reduce medical cost.

OBJECTIVETo evaluate the relation of dose intensity and efficacy, toxicity in advanced breast cancer treated with paclitaxel as a single agent.

METHODSSeventy-one patients with advanced breast cancer received paclitaxel as a single agent with different dose intensities. According to the phase I or phase II trial, the standard dose intensity of paclitaxel was defined as 58.3 mg.(m(2))(-1).week(-1). The dose of paclitaxel was 175 mg/m(2) given every three weeks, ranging 33.3 - 70.3 mg.(m(2))(-1).week(-1) [median delivered dose intensity 58.82 mg.(m(2))(-1).week(-1)]. Efficacy and toxicity was evaluated.

RESULTSThe overall response rate in this group of advanced breast cancer was 40.8%. Responses were seen in lungs, soft tissue, bone and liver, with the response rates of 52.0%, 38.0%, 12.5%, 7.7%, respectively. When the relative dose intensity (RDI) was > 1.0, 0.9 - 1.0, < 0.9, the response rates were 44.2%, 47.6%, 0, respectively. The difference between the group (RDI >/= 0.9% - 1.0%) in 7 patients and the group (RDI < 0.9) was significant (P < 0.05). Toxicity was well tolerated, with the efficacy decreased as soon as the RDI had been reduced without embarrassing the toxicity.

CONCLUSIONPaclitaxel as a single agent therapy with standard dose intensity is effective and well tolerated by patients with advanced breast cancer.

Adult ; Aged ; Antineoplastic Agents, Phytogenic ; administration & dosage ; adverse effects ; Bone Neoplasms ; drug therapy ; secondary ; Breast Neoplasms ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Humans ; Leukopenia ; chemically induced ; Liver Neoplasms ; drug therapy ; secondary ; Middle Aged ; Neoplasm Staging ; Paclitaxel ; administration & dosage ; adverse effects ; Remission Induction

OBJECTIVESTo investigate the level of AIDS knowledge among people concerned in Nanjing city in order to provide scientific evidence and constructive suggestions for the government to formulate relevant policies for AIDS control.

METHODSThree sets of questionnaires on AIDS knowledge were designed, the scores calculated, and the results evaluated.

RESULTSOf the 2,500 questionnaires issued to 4 different groups of people, 2,436 were collected back with effective answers, 991 from medical and health-related workers with the mean score of 58, 473 from college students with the mean score of 39.9, 524 from common city residents with the mean score of 42.3, and 448 from those working in high risk environment with the mean score of 47.

CONCLUSIONSThe level of AIDS knowledge among people concerned in Nanjing city was far below the requirement of the nation, especially among medical and health-related workers. Efforts must be made to raise the level of AIDS knowledge of people concerned so as to enhance the prevention and treatment of the disease.

Acquired Immunodeficiency Syndrome ; prevention & control ; Adolescent ; Adult ; China ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Risk Factors ; Surveys and Questionnaires

OBJECTIVEITS2 of DNA barcoding was used to study genetic polymorphism of Platycodon grandiflorum.

METHODTotal genomic DNA was isolated from P. grandiflorum. PCR was used to amplified the region of internal transcribed spacer 2 (ITS2), and PCR products were sequenced. The sequences of ITS2 were analyzed and compared by Clustal. The intraspecies genetic distance was calculated based on Kimura 2-parameter model by using MEGA 5.05. The ITS2 sequence of Codonopsis pilosula was used as the outreach value for plants of the genus, and the phylogenic tree used constructed by Neighbor-Joining (NJ) method.

RESULTThe K2-P's genetic distance of all samples were ranged from 0 to 0.930. The K2-P's genetic distance of samples at the same area were ranged from 0 to 0.178. The K2-P's genetic distance of samples at different areas were ranged from 0.735 to 0.930. The analytical result showed that the degree of genetic variation were heavy in intraspecies of P. grandiflorum and significantly correlated with geographical location.

CONCLUSIONThe DNA barcoding of ITS2 can applied to study the intraspecific genetic diversity, it provides a reference for further development of DNA barcoding technology applications.

China ; DNA Barcoding, Taxonomic ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Molecular Sequence Data ; Phylogeny ; Platycodon ; classification ; genetics ; Polymorphism, Genetic

OBJECTIVETo investigate the predictive value of HER-2 and ER expression for chemosensitivity of taxane in the treatment of advanced breast cancer.

METHODSOf 268 advanced breast cancer patients treated: 71 were by paclitaxel alone, 32 by docetaxel alone, 110 by paclitaxel combined with anthracylines or gemcitabine or platins and 55 by docetaxel-based combinations. HER-2 and ER expression of all patients treated by taxane underwent immunohistochemical (IHC) assay.

RESULTSUnivariate analysis showed: the response rate (RR) in HER-2 overexpression group was 56.7%, and in HER-2 weak expression group 33.3% (P = 0.003). The response rate in ER positive group and ER negative group was 33.3% and 48.9%, respectively, with a significant difference (P = 0.015). The RR was 67.6% in ER negative but HER-2 overexpression group. However, in ER positive but HER-2 weak expression group and the other groups, the RR were around 35% (P < 0. 01). Multivariate analysis showed that overexpression of HER-2 was the only significant factor to predict the chemosensitivity of taxane (P = 0. 007), but the ER, Karnofsky performance score (KPS), anthracylines, metastatic sites were not the statistically significant chemo-sensitivity predictive factors for taxane.

CONCLUSIONER negative and/or HER-2 overexpression, especially latter, may be associated with good response in advanced breast cancers treated by taxane.

Antineoplastic Agents, Phytogenic ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Multivariate Analysis ; Neoplasm Staging ; Paclitaxel ; therapeutic use ; Predictive Value of Tests ; Prognosis ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Remission Induction ; Retrospective Studies ; Taxoids ; therapeutic use

OBJECTIVETo compare therapeutic effects of warm needle moxibustion and medication on osteoporosis and to study the mechanism.

METHODSForty cases were randomly divided into an acupuncture group and a medication group, 20 cases in each group. The acupuncture group was treated by warm needle moxibustion at Dazhu (BL 11), Ganshu (BL 18), Shenshu (BL 23), Zusanli (ST 36), Yanglingquan (GB 34) etc. once other day, for 3 months; and the medication group was treated by oral administration of tablet Caltrate with Vit D2 for 3 months. The changes of bone mass density (BMD), estradiol (E2), osteocalcin (bone growth protein, BGP), urine calcium/creatinine (Ca/Cr) in the two groups before and after treatment and therapeutic effects were investigated.

RESULTSAfter treatment, BMD significantly increased (P<0.05, P<0.01) in the acupuncture group and did not signifi cantly changed in the medication group (P>0.05) with a significant difference between the two groups (P<0.05). After treatment E2 level significantly increased as compared with before treatment in both of groups (P<0.01); after treatment BGP significantly decreased as compared with before treatment in both of groups (P<0.01); after treatment Ca/Cr significantly decreased as compared with before treatment in the acupuncture group (P<0.05) ; af ter treatment, there were significant differences in BGP and Ca/Cr between the two groups (P<0.05 or P<0.01). The clinically controlled rate in the acupuncture group and in the medication group were 35.0%, 5.0%, respectively, the therapeutic effect of the acupuncture group being better than that of the medication group (P<0.01).

CONCLUSIONThe therapeutic effect of warm needle moxibustion on osteoporosis is better than that of oral administration of tablet Caltrate with Vit D2 and it can increase levels of hormones and delay bone loss. It is an effective method for preventing and treating postmenopausal osteoporosis.

Acupuncture Points ; Aged ; Bone Density ; Bone and Bones ; metabolism ; physiopathology ; Calcium ; urine ; Creatinine ; urine ; Estradiol ; metabolism ; Female ; Humans ; Middle Aged ; Moxibustion ; Osteocalcin ; metabolism ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; physiopathology ; therapy

OBJECTIVETo investigate the relative quantification of cytokeratin 19 transcription in oral squamous cell carcinoma tissues by fluorescent quantitive real-time reverse transcription-polymerase chain reaction ((RT-PCR).

METHODSCK19mRNA level was detected by fluorescent quantitive real-time RT-PCR in cancerous and para-cancerous tissues from 31 oral squamous cell carcinoma patients. According to the 2(-DeltaDeltaCt) equation, the relative quantification fold of CK19mRNA level was calculated in cancerous tissues compared with para-cancerous tissues.

RESULTSCK19mRNA levels in cancerous tissues were 2.21 folds higher than those in para-cancerous tissues, and the amplicon was specific. CK19mRNA level in cancerous tissue correlated significantly with pathological differentiation degree, the poorer the differentiation was, the higher the CK19mRNA level became.

CONCLUSIONSFluorescent quantitive real-time RT-PCR is accurate and reliable in the detection of relative quantification of CK19 transcription in oral squamous cell carcinoma tissues.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; Female ; Humans ; Keratin-19 ; genetics ; metabolism ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods