Objective:To explore the value of clinical pharmacist in clinical treatment through the pharmaceutical care on a patient with Japanese B encephalitis complicated with pulmonary infection. Methods:Clinical pharmacist participated in the whole treatment process and gave suggestions on the selection,dosage and course of the drugs prescribed for anti-virus, anti-epilepsy and anti-infection by focusing on the drug interactions and adverse reactions. Results:The treatment course of the patient was smooth, and the pathoge-netic condition was brought under control gradually while no obvious adverse drug reactions occurred. Conclusion:The work of clinical pharmacist can help to optimize treatment programs to ensure the safety and effectiveness of medication.

Objective:To explore the value of clinical pharmacists in the clinical treatment of a patient with Guillain Barré syn-drome through pharmaceutical care. Methods:Clinical pharmacists participated in the entire treatment process and gave rational sug-gestions about the selection,dosage and course of the drugs,such as glucocorticoid,immunoglobulin and antibiotics. Meanwhile,clini-cal pharmacists focused on the adverse drug reactions. Results:The patient′s condition was controlled and improved gradually. During the hospitalization,the patient developed skin rash and abnormal liver function,while the symptom was improved after the anti allergy and liver protection treatment,and no obvious effect on the primary disease was shown. Conclusion:Clinical pharmacists can help to optimize treatment regimen in order to ensure the safety and effectiveness of patients’medication.

AIM: To optimize the extraction process for Banxiaxiexin Decoction (Rhizoma pinelliae, Radix Scutellariae, Rhizoma coptidis, Radix Ginsenp, Rhizoma zinpiberis, Fruents Jujubae, and Radix Glycyrrhizae). METHODS: The content of berberine、baicalin and total solid in extract liquor were determined by orthogonal design and single factor experiment in combination with glycyrrhizic acid content and identification of Rhizoma zingiberis, Radix Ginseng and Rhizoma Pinelliae. RESULTS: The extracting was arrived at in the condition of adding eighteen times of 70% alcohol as much as crude drug and refluxing 2 times, 2h and 1h, respectively. CONCLUSION: The extraction is stable and feasible.

BACKGROUND:Previous studies have demonstrated that normal rat liver homogenate supernatant can induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels with partial hepatocyte functions. However, whether fibrotic liver homogenate supernatant can work or how the inducing effect is remains unclear. OBJECTIVE:To investigate the differentiation potential of human umbilical cord mesenchymal stem cels into hepatocytes under the normal liver and fibrotic liver microenvironment in vitro. METHODS:Liver fibrosis was induced in the SD rats by repeated intraperitoneal injections of 3% thioacetamide at a dose of 200 mg/kg body mass, twice a week for 4 weeks, and then fibrotic liver tissues and normal liver tissues were used to prepare liver homogenate supernatants. Passage 3 human umbilical cord mesenchymal stem cels were used and divided into standard control group (cels were cultured in DMEM/F12 with 10% fetal bovine serum), fibrotic liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 50 g/L fibrotic liver homogenate supernatants), normal liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 100 g/L normal liver homogenate supernatants). The morphological changes of the cels in each group were recorded under inverted microscope; the protein levels of CK18, AFP, CYP3A4, CYP2E1, CYP2D6 and TPH2 were evaluated using western blot assay. Furthermore, the concentration of albumin in the cels was measured. RESULTS AND CONCLUSION:After a 7-day inducement, the stem cels in liver homogenate supernatants groups lost their fusiform shape and changed into hepatocyte-like cels with the morphous of round shape. Compared with the standard control group, the hepatocyte-like cels in the two liver homogenate supernatants groups exhibited human hepatocyte biomarkers, CK18 and AFP. The standard control group cels could express a little amount of CYP2E1, while cels in the two liver homogenate supernatants groups could express CYP3A4, CYP2E1, CYP2D6, TPH2. Compared with the standard control group, the expression level of CYP2E1 in the two liver homogenate supernatants groups increased significantly (P < 0.01), and however, the relative levels of CYP3A4, CYP2E1, CYP2D6, TPH2 in the two liver homogenate supernatants groups showed no statistical significance (P > 0.05). At the same time, compared with the standard control group, the concentration of albumin in the two liver homogenate supernatants groups markedly increased (P < 0.01), but there was no difference between the two liver homogenate supernatants groups (P > 0.05). Experimental findings demonstrated that both of normal liver tissue and fibrotic liver tissue microenvironments could induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels. To achieve the same effect, compared with normal liver tissue, fibrotic liver tissue required lower concentrations, suggesting that fibrotic liver tissue microenvironment may be more conducive to differentiation of umbilical cord mesenchymal stem cels into hepatocytes.

Objective To select microRNAs (miRNA) associated with human cutaneous malignant melanoma (MM). Methods Total RNA was extracted from 6 tissue samples of MM and 9 human control samples of pigmented nevi, and small RNAs of less than 200 bp were enriched, miRNA microarray was used to select differentially expressed miRNAs between tissue samples of MM and pigmented nevi from 468 candi-dates. The expression of differentially expressed miRNAs was confirmed by fluorescence based real-time quan-titative PCR (qPCR) in all of these samples. Those miRNAs that were identified as differentially expressed with both miRNA microarray and qPCR were considered as significant miRNAs. Results Between the tissue samples of MM and pigmented nevi, 12.18% to 86.33% of miRNAs differentially expressed by more than 2 folds, 1.28% to 19.02% by more than 5 folds, and 0.43% to 5.34% by more than 10 folds. The expression of miRNA-21 was obviously up-regulated, while that of miRNA-320 and miRNA-494 was down-regulated in the MM samples. Conclusion There is an increase in the expression of homo sapiens miRNA-21 but a decrease in that of miRNA-320 and miRNA-494 in MM tissues.

Objective To compare three methods for the extraction of mycobacterial DNA.Methods Two commercial DNA extraction kits and an ordinary freeze-thawing method were used to extract DNA from the pure suspensions of three species of Mycobacteria (M.tuberculosis,M.leprae and M.smegmatis) at different densities (1 × 10 to 1 × 105 cells/ml),simulated clinical specimens containing different concentrations of mycobacterial cells (1 × 10 to 1 × 104 cells/ml).The purity and concentration of the extracted DNA were evaluated.Then,PCR was performed to amplify the 16S rRNA region of Mycobacteria.The performance of the three methods was compared by the purity and concentration of extracted DNA as well as the results of PCR.Further more,76 clinical skin specimens suspected to be infected with Mycobacteria were used to further validate the performance of these methods.Results All the extracted DNA samples could be detected by PCR.The highest purity of DNA was obtained by the kit A,followed sequentially by the freeze-thawing method and the kit B.When pure suspensions were used,the detection limit was consistently 1 × 102 cells/ml for all the three methods.With simulated specimens,the detection rate was consistently 100％ for all the three methods at the concentration of 1 × 103 cells/ml,60％ (12/20),55％ (11/20) and 55％ (11/20) for the kit A,kit B and freeze-thawing method respectively at the concentration of 1 × 102 cells/ml.The analysis of clinical specimens showed that the kit B could be used to extract DNA from paraffin-embedded specimens,with the detection rate similar to that of kit A and freeze-thawing method.Conclusions The kit A could rapidly yield high-quality genomic DNA of Mycobacteria by repeated cleaning of columns,and may serve as the optimal method for scientific and clinical studies,and the kit B is suitable for extracting mycobacterial DNA from fresh tissue specimens besides paraffin-embedded specimens.

Objective To discuss the experience of using the new dressings in the treatment of refractory wounds in the department of orthopedics. Methods Wound care team gave the treatment and nursing to 7 cases of refractory wounds in the department of orthopedics, used the new type of wound dressings. Results Among 7 cases, 5 cases were cured and discharged. The granulation growth of the wound was good, and the wound was healed after the transfer surgery of the joint flap. One patient had reached the healing rate of 90% after discharging, and recovered after being followed up for 22 days. Conclusions The correct assessment of the wound, reasonable choice of the new dressing, a good job of anti-infection treatment and the skin protection around the wound can effectively promote healing of the refractory wound after fracture, reduce the cost of health care, and bring the satisfaction to the patients and their families.

OBJECTIVE To study effective and convenient method for paraffin oil sterilization.METHODS By using carrier qualitative germicidal test,to compare pressure steam sterilization,dry heat sterilization and cobalt-60(gamma)-ray radiation sterilization to test the sterilizing effect and operating procedure.RESULTS Pressure steam sterilization was unable to achieve 100% sterilizing effect,whether we extended the time or use the intermittent(sterilization).After dry heat or radiation sterilization processes,no microorganism was found.CONCLUSIONS Effect of sterilization with dry heat or radiation sterilization is trustable,but its packing,operation and equipment are requested strictly,and pressure steam sterilization may be not good for paraffin oil.

Objective To investigate the predominant genotypes of β-lactamase in Klebsiella pneu-moniae isolates and the mechanism of antibiotic and histidine kinase inhibitor in affecting β-lactamase gene expression .Methods Tube dilution method and E-test were used to detect the susceptibility of K.pneumon-iae isolates to β-lactam antibiotics .The major genotypes of β-lactamase in β-lactam antibiotic-resistant iso-lates were identified by PCR and sequencing method .Disk diffusion test was performed to analyze the activity ofβ-lactamase .The effects of cefotaxime and histidine kinase inhibitor closantel on the expression of β-lac-tamase gene were evaluated by real-time fluorescent quantitative RT-PCR.Results All of the 118 β-lactam antibiotic-resistant K.pneumoniae isolates expressed β-lactamase, 90.7%(107/118) of which were KPC-2, TEM-1, CTX-M-14, SHV-11 and/or OXA-1 gene positive.The positive rates of TEM-1 (71.0%) and SHV-11 (64.5%) were significantly higher than that of the other three genotypes ( P<0.05).68.2%(73/107) of the isolates possessed two or more than two β-lactamase genotypes, and 30.8%(33/107) iso-lates were identified as TEM-1+SHV-11 genotype.Except for KPC-2 mRNA, the levels of TEM-1, SHV-11, CTX-M-14 and OXA-1 mRNA were significantly up-regulated by cefotaxime at MIC/4 (P<0.05), but were inhibited by 100μmol/L closantel (P<0.05).Conclusion TEM-1 and SHV-11 are the majorβ-lactamase genotypes carried by K.pneumoniae strains isolated from Zhejiang area , and TEM-1 plus SHV-11 ( TEM-1+SHV-11) is the predominant carrying mode of the β-lactamase genotypes .Sublethal dose of cefotaxime can enhance the expression of β-lactamase genes in K.pneumoniae, while the histidine kinase inhibitor , closan-tel, can block the increased expression of β-lactamase genes induced by cefotaxime .

Objective To optimize the concentration of a microRNA-21 (miR-21) inhibitor and a miR-494 mimic for the transfection of A375 human melanoma cells,and to estimate the effect of the miR-21 inbihitor and miR-494 mimic on the proliferation of A375 cells.Methods A miR-21 inbihitor and a miR-494 mimic were designed and constructed.To optimize the concentration of the miR-21 inbihitor and miR-494 mimic for transfection,six concentrations (70-250 nmol/L) of the inbihitor and mimic were transfected into A375 cells separately by using LipofectamineTM2000.Then,quantitative fluorescence-based PCR was performed to determine the expression of miR-21 and miR-494 in A375 cells.Some A375 cells were classified into five groups:Mock blank control group remaining untransfected,miR-21 inhibitor group transfected with the miR-21 inhibitor,miR-21 control group transfected with the miR-21 inhibitor negative control,miR-494 mimic group transfected with the miR-494 mimic,and miR-494 control group transfected with the miR-494 mimic negative control.Mter another 48-hour culture,the cells were collected for the analysis of cell apoptosis and cycle by using flow cytometry.Meanwhile,Cy5-labelled miR-494 mimic negative control was transfected into A375 cells for the evaluation of the transfection efficiency by using an inverted fluorescence microscope.Results miRNAs were successfully extracted from A375 cells.As quantitative PCR revealed,the A375 cells transfected with the miR-21 inhibitor at 120 nmol/L showed the lowest expression level (2-△△Ct) of miR-21 (average:0.80; range:0.65-0.92),and those transfected with the miR494 mimic at 250 nmol/L displayed the highest expression level of miR-494 (average:126.82; range:111.52-144.22).The transfection efficiency in A375 cells was higher than 90％.Compared with the corresponding negative control groups,the miR-21 inhibitor group and miR-494 mimic group showed increased apoptosis rate ((27.74 ± 1.39)％ vs.(12.93 ± 0.65)％,(34.30 ± 2.35)％ vs.(15.54 ± 1.02)％,both P ＜ 0.01),percentage of G1-phase cells ((61.61 ± 3.25)％ vs.(50.34 ± 5.62)％,(61.05 ± 3.17)％ vs.(49.95 ± 2.58)％,both P＜ 0.05),but decreased proliferation index ((38.39 ± 3.25)％ vs.(49.66 ± 5.62) ％,(38.95 ± 3.17)％ vs.(50.05 ± 2.58)％,both P ＜ 0.05).Conclusions Both the miR-21 inhibitor and miR-494 mimic can promote the G1-phase arrest and apoptosis in A375 cells,and miR-21 may act as a protooncogene accelerating the proliferation of A375 cells,while miR-494 may founction as a tumor suppressor inhibiting the proliferation of A375 cells.