OBJECTIVE:To determine the content of epirubicin in human plasma by RP-HPLC.METHODS:The chromatographic separation was performed on YWG C18 column.The mobile phase consisted of water-acetonitrile(12∶7,with pH adjusted to 3.90?0.05 using 85% phosophoric acid)at a flow rate of 1.0 mL?min-1.The detective wavelength was set at 254 nm.RESULTS:The linear range of epirubicin was 0.12～4.80 mg?L-1(r=0.998 4)with the lowest detective range of 0.01 mg?L-1.The intraday RSD was less than 4.40% and the inter-day RSD was less than 5.60%,and the average recovery rate was 99.40%(RSD=4.02%).The sample solution was stable within 12 h under room temperature.CONCLUSION:The method is easy,precise and reproducible to operate and the result of the determination is accurate.

Objective To discuss the methods and effects of replantation to treat complex thumb severance. Methods 56 cases of complex thumb severance combined with soft tissue defects were treated with simple vessel transplantation, veno flap transplantation, dorsal index digital island flap transplantation, digital artery transfer, transfer of radial index digital artery pedicled island flap, or veno arteriolization. The soft defects were repaired by the transfer of extensor index finger tendon and superficial ring finger flexor tendon. Results 50 cases survived and 6 cases failed, with the survival rate being 89%. 42 cases were followed up for 5 to 25 months. The function of the treated thumbs was assessed as excellent in 10, good in 25 and bad in 7. The excellent and good rate was 83%. Conclusion Complex thumb severance can be treated by unconventional replantation so as to achieve good effects.

This article analyzed the importance of implementing case teaching in health policy course based on its characteristics, and introduced the design and the process of case teaching, from the general idea, teaching content, implementation process and evaluation method. Some problems which should be paid attention to and the relevant suggestions were brought out according to the teaching practice.

Objective To explore the value of the use of the vacuum sealing drainage technology (VSD) in the dermatoplasty for large area of cutaneous defects.Methods By a prospective randomized study,64 patients with large area of cutaneous defects were randomly divided into traditional packaging group (dermatoplasty with the traditional dermatoplastic package method,n =32) and VSD group( dermatoplasty with the vacuum sealing drainage technology,n =32).Operating time,survival.rate of skin graft and wound infection were compared between these two groups.Results There was no significant difference in the operation time between the two groups( [77 ± 16]min vs [83 ± 12] min,P =0.06).The survival rate of skin graft and wound infection in VSD group were significantly better than those in traditional packaging group( survival rate of skin graft:(6±1)％ vs (20±7)％,P＜0.001;wound infection cases:1 vs6,P=0.039).Conclusion Treating large area of cutaneous defects by the dermatoplasty with VSD technology could significantly improve clinical efficacy.

Curcumae Rhizoma comes from Curcuma genus,functional breaking blood stasis,detumescence and acesodyne for treatment of Zhengjia accumulation,amenorrhea,traumatic injury and bruising pain.Modem pharmacological studies have shown that the main monomer composition of zedoary turmeric has a good anti-inflammatory and anti-tumor effects.The main monomer composition of zedoary turmeric copies of curcumol,beta etemene,curcumin anti-inflammatory anti-tumor mechanism of review,provide the basis for the further research progress and clinical application of zedoary turmeric.

Objective To investigate the diagnostic value of hysteroscopy in patients with abnormal uterine bleeding.Methods The clinical data of 547 cases of abnormal uterine bleeding were analyzed retrospectively.The diagnostic curettage was conducted after hysteroscopy,the specimens were examined by histopathology,the detection result of hysteroscopy and histopathological were compared.Results The sensitivity of hysteroscopy in diagnosed abnormal uterine cavity lesions was 95.83％ (161/168),the specificity was 97.63％ (370/379),the false positive rate was 2.37％ (9/379),the false negative rate was 4.17％ (7/168),the positive predictive value was 94.71％ (161/170),the negative predictive value was 98.14％ (370/377),the Jorden index was 0.934.Hysteroscopy diagnosis showed that the endometrial polyps was 54 cases,the pathological diagnosis coincidence rate was 79.63％ (43/54);93 cases of simple hyperplasia,the pathological diagnosis coincidence rate was 90.32％ (84/93);16 cases of atypical hyperplasia,the pathological diagnosis coincidence rate was 87.50％ (14/16);4 cases of differentiated endometrial adenocarcinoma,the pathological diagnosis coincidence rate was 100.00％ (4/4);2 cases of uterine papillary serous carcinoma,the pathological diagnosis coincidence rate was 100.00％ (2/2);1 case of transitional cell carcinoma of endometrium,the pathological diagnosis coincidence rate was 100.00％ (1/1).Conclusion Hysteroscopy can visually observe the morphology of cervical canal and uterine cavity,the diagnostic rate of endometrial polyps,simple hyperplasia,atypical hyperplasia and endometrial carcinoma are higher,so it has important clinical value in the diagnosis of abnormal uterine bleeding.

Background Krüppel-like factor 6 (KLF6) is related to the physiological or pathological process,such as growth,cell differentiation,proliferation,apoptosis,angiogenesis,tissue repair,and so on.But in ophthalmology,it is less reported about the expressing level of KLF6 protein in lens epithelial cclls (LECs) or the effect of KLF6 on the proliferation of human LECs.Objective This study was to investigate whether KLF6 can inhibit proliferation of human LECs.Methods KLF6 eukaryotic expression plasmid (pEGFP-C2-KLF6) was constructed using reverse transcription PCR(RT-PCR) and identified by double enzyme digestion method and PCR.Human LECs strain (HLE-B3) was cultured and passaged using low glucose DMEM containing 10％ fetal bovine serum and then divided into 4 groups.KLE-B3 transfection reagents were added in the culture medium of all groups.In addition,no agent was used in the blank control group;only insulin-like growth factor-1 (IGF-1) was appended to the medium in only IGF-1 group ;null vector was transfected and IGF-1 was appended in the null plasmid transfection+ IGF-1 group;while pEGFP-C2-KLF6 eukaryotic expression plasmid was transfected into the cells,and simultaneously IGF-1 was added in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group.After 24 hours of intervene,water soluble tetrazolium salt-1 (WST-1) test was used to detect the growth status of the cells,and Western blot assay was used to assay the relative expressing level of KLF6 protein in the cells.In the other hand,the cells were cultured at the density of 1 ×104/piece,and 0,0.10 and 0.25 μg pEGFP-KLF6 were transfected into each piece of cells respectively,and then IGF-1 was added with a final concentration of 50 μg/L for 24 hours after cell culture.Expressions of Ki-67 protein and mRNA in the cell pieces were detected by immunocytochemistry and fluorescent semiquantitative PCR,respectively.Results The PCR product bands were consistent with KLF-6 gene in length,and the product fragments were corresponding with expectant ones via PCR and double enzyme digestion method,showing a successful construction of pEGFP-C2-KLF6 eukaryotic expression plasmid.After 24 hours of IGF-1 stimulation,the absorbance values of the cells were 0.86±0.00,2.10±0.01,2.24±0.12 and 1.06±0.02 in the blank control group,only IGF-1 group,null plasmid transfection + IGF-1 group and the pEGFP-C2-KLF6 plasmid transfection + IGF-1 group,with a significant difference among the 4 groups (F =38.322,P ＜ 0.05),and that in the pEGFP-C2-KLF6 plasmid transfection +IGF-1 group was significantly lowed in comparison with the only IGF-1 group and the null plasmid transfection+IGF-1 group (q=6.42,7.31,both at P＜0.05).Western blot assay showed that the relative expressing levels of KLF6 protein were statistically different among the four groups (F =591.858,P＜0.05),and those in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group were 1.47,2.04,3.27 folds higher than those in the blank control group,only IGF-1 group,respectively and null plssmid transfection+IGF-1 group,respectively.Immunocytochemistry revealed that the expressing intensity of Ki-67 protein was gradually weakened with the decrease of pEGFP-C2-KLF6 plasmid dose in the 0,0.10 and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+IGF-1 groups.Fluorescence semiquantitative PCR results showed that the relative expression values of Ki-67 mRNA in the cells were O.15±0.08 and 0.11±0.03 in the 0.10 μg and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+ IGF-1 groups,which were significantly lower than O.77± 0.12 of the 0 μg pEGFP-C2-KLF6 plasmid transfection group,with a statistically significant difference among the three groups (F =54.825,P＜0.05).Conclusions KLF-6 can effectively inhibit IGF-1-induced proliferation of human LECs,and it can be regarded as one of the regulatory factors of the proliferation of HLE-B3 cells.

Objective To explore the writing quality and rational use of drugs for outpatient prescription of western medicine in Yanqing District Hospital of Beijing.Methods From January 2011 to December 2015, 12 000 prescriptions of western medicine were selected randomly.The prescriptions were analyzed by Excel and the Pareto diagram analysis was used to find out the main reasons of irrational prescription.Results From 2011 to 2015, the basic indicators of prescription of western medicine were reasonable:the average number of drugs on each pre-scription was 2.31,the utilization rate of injections was 15.91%,the utilization rate of essential drugs was 38.00%, the utilization rate of antibacterial was 14.56%.The unreasonable rate of prescription was 6.32%,and the main irra-tional reason were dosage inappropriate,indications inappropriate,inappropriate selection of drugs and combination therapy was not suitable.Conclusion By prescription review,we have listed the problems,and provide the basis for rational drug use and improvement measures.

Objective To investigate the humoral and cellular immune responses in patients with acute brucellosis, and evaluate dynamic changes of regulatory T-lymphocytes (Foxp3+ Treg) in the peripheral blood of patients during treatment, in order to clarify the relationship between immunosuppression and the therapeutic effect in human brucellosis.Methods Sixty-five patients with brucellosis hospitalized at the Third Department of Infectious Diseases, Heilongjiang Agriculture and Reclamation Bureau General Hospital between July 2015 and November 2015 were included.Twenty-eight patients were treated with conventional therapy (group A: patients received 3 courses of treatment.Each lasted for 20 days with one-week interval), and 37 patients were treated with conventional therapy in combination with immunopotentiator (group B).Thirty healthy volunteers were enrolled as the controlled group.The ratio of CD3+CD4+ Foxp3+ Treg cells in the peripheral blood of brucellosis patients were measured by flow cytometry (FCM) at the end of each course of treatment.Data in accordance with normal distribution were described as mean±standard deviation.Comparison between two groups was done by two sample t test.Comparison among multiple groups was performed by analysis of variance and SNK test.Data that did not fit the normal distribution were analyzed by multiple-sample nonparametric test.Results After the first (20 d), second (50 d) and third course of treatment (80 d), the ratios of Foxp3+Treg in the peripheral blood of 65 acute brucellosis patients were 2.83%, 3.77% and 4.03%, respectively, which were all significantly higher than control group (1.69%;t=5.97, 9.05 and 5.66, respectively, all P<0.01).At the end of the first course of treatment, the ratios of Foxp3+Treg in group A and B showed no statistically difference (t=0.33, P>0.05), while those were both higher than control group (t=7.09 and 4.94, respectively;both P<0.01).At the end of the second course, the ratio of Foxp3+ Treg in group B was higher than group A (t=2.22, P<0.01), and both of them were higher than control group (t=10.79 and 7.25, respectively;both P<0.01).At the end of treatment, Foxp3+ Treg in group A was also significantly higher than the other two groups (t=6.02 and 6.45, respectively;both P<0.01).Conclusions In patients with acute brucellosis treated with the standard antibiosis treatment in combination with immunopotentiator, the ratio of Foxp3+Tregs significantly increases and maintains at a high level, which suggests that extra immunopotentiator may be not helpful for the treatment of brucellosis at the very early stage.

OBJECTIVETo investigate the effect of functional blocking of endogenous miR-23a with a specific antisense oligonucleotide (ASO) on the proliferation and invasiveness of gastric adenocarcinoma cell line MGC803 in vitro.

METHODSA specific ASO targeting miR-23a, namely ASO-23a, was transfected into MGC803 cells to block endogenous miR-23a. The mRNA level of miR-23a in the transfected cells was detected with quantitative real-time PCR. The changes of cell proliferation following the transfection were detected with MTT assay and colony formation assay, and TUNEL assay and Transwell assay were employed to evaluate the changes in cell apoptosis and invasiveness, respectively.

RESULTSQuantitative real-time PCR demonstrated efficient functional blocking of endogenous miR-23a in MGC803 cells by ASO-23a. Suppression of miR-23a with ASO-23a obviously inhibited cell growth, colony formation and invasiveness of MGC803 cells and significantly enhanced the cell apoptosis.

CONCLUSIONASO-23a can efficiently block the function of endogenous miR-23a in MGC803 cells to inhibit cell proliferation and invasion and promote cell apoptosis.

Adenocarcinoma ; genetics ; pathology ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; MicroRNAs ; genetics ; Oligonucleotides, Antisense ; Stomach Neoplasms ; genetics ; pathology ; Transfection