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.

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 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.

Objective To study CT findings and prognosis of hypoxic -ischemic encephalopathy in full term neonates .Methods Two hundred and sixty-eight cases of full-term neonates diagnosed clinically as hypoxic-ischemic enceohathy(HIE) were examined.Seventy-nine cases were observed by CT follow-up one or four times.Nineteen normal neonates with same age were selected as contral group.Results Two hundred and thirteen cases(79%)were abnormal on CT.The CT value of the low density area of brain white matter were lower than 18 HU in two hundred cases(94%).The low density area were enlarged in one hundred and ninty two cases(90%).Subarachnoid hemorrhage(SAH) were found in seven cases(3.3%) and intraventricular hemorrhage(IVH) in four cases(2%), and infarction in one case (0.5%).CT follow-up study of 51 cases were carried out around one year of age . 27 of them were abnormal on CT,mostly showing external hydrocephalus,minority of them with cerebral atrophy and softening .23 cases were studied by CT follow-up at or after two years old,3 of them had external hydrocephalus and 1 of them had cerebral softening.Conclusion CT findings of HIE mostly had :①brain edema showing lower density area of enlarged brain white matter;②brain hemorrhage mostly showing subarachnoid hemorrhage and intraventricular hemorrhage;③seldom brain infarction.Prognosis of HIE which was mild and moderate degree based on CT findings was better ,and severe degree was bad.

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 symptoms and affecting factors of food intolerance among 903 adults.Methods Food special antibody(IgG)in human serum were examined by ELISA.Food Intolerance Health Assessment Questionnaire was used to assess positive rate of food intolerance and food intake habits.Chi-square test and Mann-Whitney test were used for data analysis.Results The ratio of food intolerance was 0.33％ to 28.13％.Intolerant foods include crab(28.13％),yolk(27.02％),milk (13.07％),soybean(11.96％),shrimp,tomato,corn,beef,rice,chicken,mushroom,wheat,and pork.The symptoms of nervous system(55.89％),digestive system(45.69％)and respiratory system(15.64％)were commonly seen.Prevalence of food intolerance in mental labors was lower than manual labors.Conclusion The most common intolerant food includes crab,yolk and milk.The symptoms of food intolerance are different.Manual labors are more susceptible to food intolerance.

Objective The treatment of limb bone fracture in combination with main vessel injury wasanalyzed retrospectively.Methods Twenty ccsese were respectively conducted by interlocking intramedullary pin fixation,plate and screw fixation,plate and screw fixation orextemal brace fixation according to the injury conditions of the fractures.Then,vascahr injaries were flexibly dwelt with direct repair,end-to-end anastomosis or blood vessel grafting.Results The-limb save rate in this study is 70%.and the reaoons which csllsed the amputation included:Long ischemia time and serious tissue damage.Conclusion Fractures of the extremities with major vascular injuries should be diagnosed promptly to be conducted properly.In this way,better outcome would be obtained.

Objective To investigate the practical anatomy and clinical applications of the lateral sural nerve-lesser saphenous vein vascular plexus island flap with median sural nerve preserved.Methods Ten lower limbs injected with color red latex and 20 fresh limbs were observed.Based on anatomical studies,the island flap pedicled with the vascular axis of lateral sural nerve and lesser saphenous vein was designed for clinical application in 63 cases.In procedure,the median sural nerve were preserved in donor site and the distal end of the lesser saphenous vein were not ligated.Results The accompanying arteries formed a favourable vascular plexus around the nerve and the vein,lied along the lateral sural nerve and the lesser saphenous vein and nourished the skin through cutaneous branches.The distance of the lateral ankle to the confluent of lateral and median sural nerve was (8.5± 0.8) cm.63 patients with soft tissue defects in the distal leg and foot were repaired.Their appearance and function were satisfaction without feeling lose in the heel and lateral side of the foot.Conclusions The lateral sural nerve-lesser saphenous vein vascular island flap is a reliable source of soft tissue to cover defects in the lower leg and foot.This flap has sufficient blood supply and high survival rate without the sacrifice of the median sural nerve.

Objective To evaluate the application of modified Abbe flap in repairing moderate defects of the upper lip and time point to divide the pedicle.Methods Classic Abbe flap was modified in its design,pedicle cutting and dividing time,which was used to repair moderate defect of the upper lip in 12 cases.Surgery was divided into two phases:one with modified Abbe flap surgery was performed for the combined nasal deformity repair simultaneously,and then the pedicle was divided 9days after surgery.Results 12 patients underwent modified Abbe flap.No vascular complications were found in these flaps.Upper lip shape was well and satisfactory functional recovery,corresponding improvement in nasal appearance.Conclusions The surgery that the modified Abbe flap with the pedicle is divided 9 days after surgery is very simple.On one hand,it greatly improves the patient's appearance and function of the upper lip,improve the overall shape of midface,on the other hand,dividing pedicle time is significantly shorter than in the past,specially reducing the suffering of patients and duration.It is particularly suitable for unilateral and bilateral cleft lip of the upper lip on secondary moderate deformities and combined nasal deformities.