Microwave-assisted extraction (MAE) was used for extraction of effective components of sarcandra glabra (Thunb.), and then chromatographic fingerprint of sarcandra glabra (Thunb.) was studied by high performance liquid chromatography/diode array detector (HPLC/DAD). The conditions of MAE were optimized by an orthogonal experiment, and then the authentication and validation of the chromatographic fingerprint were conducted. Nine peaks were identified as common peaks in the fingerprint chromatograms, and isofraxidin was considered as a reference compound and quantified. Relative standard deviations of retention time and peak area of each component were less than 3 % and 8 %, respectively. Similarity and difference analysis were conducted by use of PCA and relation coefficient. Twenty batches of sarcandra glabra (Thunb.) samples from two different producing areas could be classified into two different groups in the PCA model. The results showed that MAE-HPLC/DAD method was simple, efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra (Thunb.), which could provide more reliable and precise information for quality evaluation.

Microwave-assisted extraction (MAE) was used for extraction of effective components of sarcandra glabra (Thunb.), and then chromatographic fingerprint of sarcandra glabra (Thunb.) was studied by high performance liquid chromatography/diode array detector (HPLC/DAD). The conditions of MAE were optimized by an orthogonal experiment, and then the authentication and validation of the chromatographic fingerprint were conducted. Nine peaks were identified as common peaks in the fingerprint chromatograms, and isofraxidin was considered as a reference compound and quantified. Relative standard deviations of retention time and peak area of each component were less than 3 % and 8 %, respectively. Similarity and difference analysis were conducted by use of PCA and relation coefficient. Twenty batches of sarcandra glabra (Thunb.) samples from two different producing areas could be classified into two different groups in the PCA model. The results showed that MAE-HPLC/DAD method was simple, efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra (Thunb.), which could provide more reliable and precise information for quality evaluation.

Objective T o analyse the m etabolic changes in urine of rats w ith brodifacoum intoxication, and to reveal the m olecular m echanism of brodifacoum-induced toxicity on rats. Methods B y establish-ing a brodifacoum poisoning rats m odel, the urine m etabolic profiling data of rats w ere acquired using high performance liquid chromatography-timeofflightmassspectrometry (HPLC-TOF-M S).The orthogo-nal partial least squares analysis-discrim ination analysis (O PLS-D A ) w as applied for the m ultivariate statistics and the discovery of differential m etabolites closely related to toxicity of brodifacoum . Results O PLS-D A score plot show ed that the urinary m etabolic at different tim e points before and after drug adm inistration had good sim ilarity w ithin tim e period and presented clustering phenom enon. C om paring the urine sam ples of rats before drug adm inistration w ith w hich after drug adm inistration, tw enty-tw o m etabolites related to brodifacoum-induced toxicity w ere selected. Conclusion T he toxic effect of brodi-facoum w orked by disturbing the m etabolic pathw ays in rats such as tricarboxylic cycle, glycolysis, sphin-golipid m etabolism and tryptophan m etabolism , and the toxicity of brodifacoum is characterized of accu-m ulation effect. The m etabonom ic m ethod based on urine H PLC-TO F-M S can provide a novel insight into the study on m olecular m echanism of brodifacoum-induced toxicity.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Multiple Organ Failure ; complications ; drug therapy ; prevention & control ; Phytotherapy ; Systemic Inflammatory Response Syndrome ; complications ; drug therapy ; prevention & control

Multidrug resistance proteins (MRP2, ABCC2) may play a role in drug resistance in epilepsy by limiting gastrointestinal absorption and brain access of antiepileptic drugs (AEDs). We sought to investigate the effects of ABCC2 polymorphisms on plasma carbamazepine (CBZ) concentrations and pharmacoresistance in Chinese patients with epilepsy. ABCC2 rs717620, rs2273697, rs3740066 polymorphisms were genotyped by polymerase chain reaction amplification followed by restriction fragment length polymorphism analysis or direct automated DNA sequencing in 80 patients treated with CBZ monotherapy. There were no differences in CBZ maintenance doses or adjusted plasma CBZ concentrations among the ABCC2 rs717620, rs2273697 and rs3740066 genotypic groups. No associations between all the studied genotypes and haplotypes involving the three SNPs of ABCC2 and CBZ resistance were observed in this patient cohort. These results suggest that ABCC2 polymorphisms may not contribute to interindividual variabilities in CBZ daily maintenance doses, plasma concentrations, and treatment efficacy.
Epilepsy

Objective To build the experimental basement for the clinical use of cytokines induced killer(CIK)cells from the cord blood mononuclear cells(CBMNC)in tumor adoptive cellular immunotherapy, an effective protocol for their proliferation in vitro and cytotoxicity of CIK cells was established.Methods The lymphocytes from umbilical cord blood were isolated by density gradient centrifugation and suspended in medium with CD_3 mAb,rIL-2,rIL-1 and IFN-? as inducing agents to prepare CIK cells.At the same time, the lymphokine activated killer(LAK)and CBMNC were set as controls,which were only added IL-2 and not any cytokines during the whole culture.The changes of CIK cells before and after induction were observed with microscope and the phenotypes of the cells were analyzed by using flow cytometry.The proliferation of CIK cells were determined by trypan blue exclusion assay and the cytotoxic activity to lung cancer cell were tested with MTF method.Results According to the experiment,combining use of four types of cytokines could generate a great deal of CIK cells possessing highly cytotoxicity.From day 5 CIK cells became to prolif- erate and reached the peak at day 14.During the whole period,the relative percentage of CD_3~+ CD_(56)~+ cells in- creased significantly.Compared with LAK cells,which reached the proliferation peak at day 7 and then showed no evident proliferation.The control cells(CBMNC)showed no evident change of phenotypes and proliferation.CIK cells showed a higher antitumor activity on the tumor cells than LAK cells and CBMNC in vitro.Conclusion Umbilical cord blood can generate a great deal of CIK cells combining used with cy- tokines.Compared with classic LAK cells,umbilical cord blood CIK cells have the advantages of rapid prolif- eration speed and powerful cytotoxicity.CIK cells will be promising as a new strategy for the adoptive cellular immunotherapy of tumor.

To investigate the clinicopathological features of endometrial endometrioid adenocarcinoma(EEA)with adnexal involvement.Methods The clinicopathological data of 86 EEA patients who underwent surgical treatment at our center from January 2000 to December 2015 were analyzed retrospectively.The clinicopathological features were compared between patients with occult adnexal involvement and those with gross adnexal involvement.Results A total of 86 EEA patients with adnexal involvement(mean age:58.1 years)were included in this study,accounting for 5.4%(86/1592)of the EEA patients during the same period.Among these 86 patients,there were 13 premenopausal patients(15.1%)including 2 premenopausal patients aged under 40 years.Gross adnexal involvement was found in 47 patients(54.7%),while occult adnexal involvement was found in 39 patients(45.3%)in pathology evaluation.Ovarian metastasis was found in 34 patients(39.5%),followed by both ovarian and tubal metastasis in 19 patients(22.1%)and tubal metastasis in 33 patients(38.4%).The expressionss of estrogen receptor(χ=8.086,P=0.042)and progesterone receptor(PR)(χ=9.149,P=0.026)were significantly different between gross adnexal involvement group and occult adnexal involvement group,whereas no significant difference was found in other clinicopathological features(all P>0.05).The non-conditional Logistic regression analysis showed that,compared with PR no-expression group,the rate of occult microscopic adnexal involvement in PR low-expression group was 6.375 times of that of the gross adnexal involvement(P=0.005,95%CI:1.768-22.976),and the rate of occult microscopic adnexal involvement in the PR high-expression group was 3.719 times of that of gross adnexal involvement(P=0.048,95%CI:1.009-13.702). Conclusion PR expression level is remarkably lower in EEA patients with gross adnexal involvement than those with occult adenxal involvement.

By retrospectively reviewing the current status of traditional Chinese medicine (TCM) and Western medicine treatments of chronic nonbacterial prostatitis (CNP), the TCM understanding of its etiologies and pathogenesis, the therapeutic principles and the mechanisms of acupuncture treatment of CNP, the clinical study strategies of acupuncture treatment for CNP were further proposed, which could provide more scientific basis and support for the definite longer-term therapeutic efficacy of acupuncture treatment of CNP. Breakthrough in the treatment of CNP will be achieved with the application of acupuncture therapy both in clinical practice and experimental research.

SUMMARY To explore the clinical pathological characteristics and improve the recognition in the diag-nosis and treatment of basal cell carcinoma (BCC)of prostate.Three cases of BCC of prostate were re-ported and the relevant literature was reviewed to investigate the diagnosis and treatment of this disease. We analyzed three cases of prostatic BCC.Their ages were within a range of 57 to 83 years.One of them complained of hematuria and two complained of dysuria.All of them presented with prostatic hyperplasia. Two of them presented with high prostate specific antigen (PSA)and one with normal PSA.Case 1 had prostate cancer invasion of bladder,rectal fascia,with lymph node metastasis,bone metastasis and lung metastases.The patient received bladder resection +bilateral ureteral cutaneous ureterostomy +lymph node dissection on November 2,2014 .Postoperative pathological diagnosis showed BCC.Reexamination of pelvic enhanced MRI in January 8,2015 suggested pelvic recurrence.Abdominal enhanced CT showed multiple liver metastases and pancreatic metastasis on July 11,2015.Prostate cancer specific death occurred in October 2015.Case 2 was diagnosed as BCC in prostate biopsy on March 27,2015. Positron emission tomography and computed tomography (PET-CT)showed pulmonary metastasis and bone metastasis.Then the patient received chemotherapy,endocrine therapy and local radiation therapy. Reexamination of PET-CT on January 11,2016 showed that the lung metastase tumors and bone metas-tase tumors were larger than before.Up to January 10,2016,the patient was still alive.Postoperative pathological changes of transurethral resection of prostate (TURP)in case 3 showed BCC might be con-sidered.The PET-CT suggested residual prostate cancer,which might be associated with bilateral pelvic lymph node metastasis.In April 20,2016,the review of PET-CT showed pelvic huge irregular hybrid density shadow,about 14.5 cm ×10.0 cm ×12.9 cm in size,and tumor recurrence was considered. Then the patient received local radiation therapy.The patient survived in the followed upon January 10, 2016.BCC of prostate is a rare subtype.Due to the local infiltrative and distant metastatic potentiality, active management is preferred and a life-long follow-up is necessary.

Enhanced green fluorescent protein( EGFP), myc epitope and polyhistidine metal-binding tag are often used as a marker for recombinant fusion protein in many gene expression vectors, each marker has its own function, EGFP emits green fluorescence for direct detection, myc epitope facilitates recombinant fusion protein detection using its antibodies, polyhistidine tag allows purification of recombinant fusion protein using resin.Hitherto, no a plasmid vector can integrate all of these functions. In this study we constructed a novel eukaryotic expressive plasmid, designated as pcDNA6/myc-his-EGFP B, which integrated the functions of EGFP, myc epitope and polyhistidine tag. Importantly, a linker octo - peptide in N terminal of EGFP was designed using LINKER program. A DNA fragment encoding a putative protein containing a signal peptide of human interleukin 2(IL-2) in N terminal was cloned into pcDNA6/myc-his-EGFP B in frame with the C-terminal peptide to construct pMHES. 2.2.15 Cells were transfected with pcDNA6/myc-his-EGFP B and pMHES, and Balb/c mice were intravenously injected with pcDNA6/myc-his-EGFP B by tails, results revealed that both of the plasmids worked in 2.2.15 Cells and livers of Balb/c mice. Assuming gene of the IL-2 was inserted into pcDNA6/myc-his -EGFP B in frame with EGFP, myc and 6 × His, three-dimensional structure for this putative expression product was simulated using Modeller8V2, results revealed that IL-2, EGFP, myc and 6 × his did not interfere each other and octo- peptide linker owned certain flexibility. The results suggest that pcDNA6/myc-his-EGFP B may be useful as a genetic tool for mammalian cells and a vector for gene therapy.