OBJECTIVE:To provide reference for further implementation and improvement ofTwo Tax Billmedicines distri-bution reform. METHODS:Literature review and field investigation were used to systematically sort out,summarize and analyze the effect ofTwo Tax Billmedicines distribution reform and potential problems. Corresponding policy suggestions were proposed. RESULTS & CONCLUSIONS:TheTwo Tax Billmedicines distribution reform boosted the integration and re-combination of medicines distribution system,and effectively rectify the medicines distribution system. However,this study found no evidence of its effect on reducing the high medicines prices. We suggest that,the implementation ofTwo Tax Billmedicines distribution re-form should firstly ensure the stable supply of medicines,especially the availability of essential medicines in rural remote areas, and those medicines with high risk of shortage. It is also suggested to strengthen the audit of tax receipts;establish long-term mecha-nism to promote on-time payment to suppliers by hospitals;break down local protection;set up efficient multi-ministerial coordina-tion mechanism.

Objective To investigate clinical predictors in early pregnancy for adverse pregnancy outcomes in women complicated with chronic nephropathy. Methods One hundred and eighty-four pregnancies complicated with chronic nephropathy who delivered between January, 2005 and January, 2014 in Peking University First Hospital were retrospectively analyzed. Two hundred pregnant women without chronic nephropathy were selected as the control group. Pregnancy outcomes were compared between the two groups. Relationships between clinical predictors in early pregnancy [age, history of kidney disease, drugs used before pregnancy (in three months), results of renal biopsy, blood pressure, serum creatinine, urea nitrogen, 24 h urinary protein, estimated glomerular filtration rate, stage of chronic nephropathy] and adverse pregnancy outcomes [maternal mortality;pregnancy complicated by severe preeclampsia;renal function decline, early preterm birth, very low birth weight infants, fetal loss after 20 weeks of pregnancy] were assessed by logistic regression analysis. T-test, Chi-square test and multivariate regressions were used for statistical analysis. Results There were 174 and 197 livebirths in the study and the control groups, respectively. The most popular type of chronic nephropathy in pregnant women was IgA nephropathy (38.6%, 71/184). Compared with the control group, the risk of premature labor [9.8%(17/174) vs 3.0%(6/197), χ2=7.184], low birth weight infants[18.4%(32/174) vs 7.1%(14/197),χ2=9.813], very low birth weight infants [5.7%(10/174) vs 1.5%(3/197),χ2=4.536], fetal loss after 20 weeks of pregnancy [5.7%(10/184) vs 1.5%(3/200), χ2=4.536] and severe preeclampsia [17.9%(33/184) vs 1.5%(3/200), χ2=33.544] increased significantly in the study group (all P < 0.05). Twenty-four-hour urinary protein content (OR=1.84, 95%CI: 1.36-2.50, P=0.001) and mean arterial pressure (OR=1.04, 95%CI: 1.00-1.07, P=0.027) in early pregnancy were risk factors for adverse pregnancy outcomes. The risk of renal function decline increased significantly in patients with higher stages of chronic nephropathy in early pregnancy (OR=6.50, 95%CI: 3.34-8.21, P<0.01). Mother complicated by preeclampsia during pregnancy was an independent risk factor (OR=11.10, 95%CI: 4.48-27.20, P<0.01). Compared with women whose 24 h urinary protein content less than 1g in early pregnancy (122 livebirths within 126 cases), the risk of premature labor [17.3%(9/52) vs 6.6%(8/122), χ2=4.780], increased significantly in women whose 24 h urinary protein content were more than or equal to 1 g in early pregnancy (52 livebirths within 58 cases) (P<0.05). Conclusions Elevated urinary protein level and mean arterial pressure in early pregnancy were risk factors of adverse pregnancy outcomes. The risk of renal function decline increased in patients with higher stages of chronic nephropathy in early pregnancy. Mother complicated by preeclampsia during pregnancy was an independent risk factor for poor prognosis of the fetus.

Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

AIM:To investigate the potential of hematopoietic stem cells(HSCs)derived from mouse embryonic stem cells(ESCs)to reconstruct hematopoiesis in vivo.METHODS:Using a three-step method,a mice embryonic stem cell line,E14.1 was induced into hematopoietic stem cells.The cell markers with CD34+/Sca-1+ were identified by flow cytometry analysis,then HSCs(1?109 cells/L)from third-step were injected into SCID mice for observing teratoma formation.To validate function of HSCs,colonogenic cell assay was conducted and the hematopoiesis in lethally irradiated mice was reconstituted.RESULTS:The method of three-step differentiation,combined to use more hematopoietic stimulating factor promoted the E14.1 cell differentiation into HSCs with highest percent of CD34+/Sca-1+ cells(as high as 58.64%?4.20%)with more CFU-E,CFU-GM and CFU-GEMM populations.The cells showed the character of hematopoietic progenitors by Wright-Giemsa staining.Positive selected CD34+/Sca-1+ cells by magnetic sorting from third-step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 d after transplantation,with 71.4%(5/7)successful engraftment rate.Three recipients showed that the cell population of the peripheral blood leukocytes,red blood cells and hemoglobin approached to normal index at 40 d after transplantation,but followed relative slow renew in platelet count.Survival rate in transplant group was 43%,compared to 100% mortality in control mice.Karyotyping assays confirmed the female mice with XY.CONCLUSION:The three-step differentiation and the culture conditions described here support the differentiation of mouse ESCs into HSCs.HSCs derived from mouse ESCs can reconstruct hematopoiesis.

Objective To evaluate the effect of pressure load of avascular necrosis of the femoral head(ANFH) by percutaneous injection of polymethylmethacrylate(PMMA).Methods Models of ANFH were established in 20 adult white rabbits with 40 femoral heads which were induced by injection of hydrochloride(HCL) and hormone.DR was performed to observe the bony density of femoral head in all rabbits,the cystic degeneration and collapse of femoral head were detected in 21 femoral heads,then DSA was done to observe the blood supply and they were divided into three groups at random,group A in 5 was examined by pathology,group B in 8 was injected with PMMA and group C was as control.The test of pressure loading was performed in group B and C.Results Of 21 femoral heads with cystic degeneration and collaps,8 were in the HCL-induced group and 13 were in the hormone-induced group.DSA showed that the blood supply was decreased,and the arteria were decreased in diameter.After the percutaneous injection of PMMA,all of 8 rabbits in experimental group were successful.The distinction between the means of two samples was significant in loading test(P

AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.

Objective To investigate the possible effect of ginsenoside Rg1 and oligo Aβ1-42 on PKA/CREB pathway.Methods The damage was induced by oligomeric Aβ1-42 in primary cortical neuron.Neurons were incubated with or without glutamate,or incubated in Aβ,or pre-incubated in Rg1 and then co-incubated in Aβ.The proteins of p-CREB,t-CREB,PKA Ⅱ α and BDNF were detected by Western blot.Results After the treatment with Oligo Aβ1-42 for 2 h,the p-CREB/t-CREB level induced by glutamate was obviously lower (P＜ 0.001).However,in neurons pre incubatedwith 2.5,5.0,10.0 μmol/L of ginsenoside Rg1 and then co-incubated with 5μmol/L of oligo Aβ1-42,the p-CREB/t-CREB induced by glutamate was significantly increased as compared with that of Aβ1-42 group (P＜0.05).Upon Aβ1-42 exposure for 2 h,cortical neurons showed a statistically significant increase in PKA Ⅱ α as compared to the control group (P ＜ 0.001).Pre-treatmentwith varying doses of ginsenoside Rg1 (2.5,5,10μmol/L) showed a decrease in PKA Ⅱ α as compared to neurons treated with Aβ1-42 alone for 2 h (P＜0.001).Furthermore,BDNF level significantly increased in Rgl-pretreated cells as compared to cells treated with Aβ1-42 alone for 24h (P＜0.05).Conclusions Ginsenoside Rg1 attenuates the oligo Aβ142 inhibition of PKA/CREB pathway.

This paper analyzes and discusses the research advances of acupuncture in the treatment of childhood cerebral palsy from the perspectives of clinical treatment and its mechanism. Acupuncture has great advantages in the treatment of childhood cerebral palsy,however, there are methodological problems in current researches, clinical researches in particular. High-quality clinical researches should be conducted to improve clinical efficacy fully.

DPP-4 inhibitors are new oral hypoglycemic drugs and hot spots developed and launched in recent years, and they pro-vide new choices for the clinical treatment of type 2 diabetes. In China, DPP-4 inhibitors that are approved to use in the treatment of type 2 diabetes are all imported products currently. In the paper, the current intellectual property situation of DPP-4 inhibitors that are developed and approved at home and abroad is researched and analyzed. Reasonable use of the patent information of DPP-4 inhibitors that is about to expire or have failed can provide good guidance for the subsequent development of DPP-4 inhibitors in domestic with promising curative effect and good market prospects, and can generate new patents in order to enhance the market competitiveness.

Aim To explore the possible protective effect of ginsenoside Rg1 on oligomeric Aβ1-42 induced apoptosis and its possible mechanism. Methods The damage was induced by oligomeric Aβ1-42 in primary cortical neurons. Cells were incubated in the absence or presence of Aβ, or co-incubated in sp600125 with Aβ, or pre-incubated in ginsenoside Rg1 then co-incu-bated in Aβ. The p-JNK, JNK, caspase-3 activity and TUNEL-positive cells were detected. Results In Aβ1-42 treated group, the ratio of p-JNK/JNK level was increased more than that in non-treated group for 15 min. However, in neurons preincubated with (2. 5, 5, 10 μmol·L-1 ) ginsenoside Rg1 and then co-incuba-ted with 5 μmol·L-1 oligomeric Aβ1-42 , the p-JNK/JNK ratio, caspase-3 activity and TUNEL positive neu-rons were significantly decreased compared with those of Aβ1-42 treated group. Conclusion Ginsenoside Rg1 can attenuate the oligomeric Aβ1-42-induced apop-tosis by JNK pathway.