BACKGROUND: There are currently many cryopreservation methods for the aminotic membrane, which have varying effects on the ultrastructure and biological activity of amniotic membrane, but on no one is effective.
OBJECTIVE: To compare the effects of different cryopreservation methods on the ultrastructure and viability of aminotic membrane and to seek the ideal cryopreservation method.
METHODS: Aminotic membrane separated from the fresh placenta was preserved respectively with deep-frozen cryopreservation and vitrification, and everyway was run for 3 and 6 months. Fresh aminotic membrane was used as control. The ultrastructure of aminotic membrane was observed by transmission electron microscopy, and the viability of aminotic membrane was assessed by microcomputer analysis system for biological oxygen consumption, and immunohistochemical staining combined with image analysis system was used for lactate dehydrogenase activity.
RESULTS AND CONCLUSION:After 3 and 6 months of crypreservation, the damage to the ultrastructure of aminotic membrane by vitreous cryopreservation was slighter than that of amniotic membrane cryopreserved at-80℃. Compared with the fresh aminotic membrane, the gray value of lactate dehydrogenase and partial pressure of oxygen were significantly decreased in the cryopreserved aminotic membrane by deep-frozen cryopreservation at 3 and 6 months (P < 0.05) and by vitreous cryopreservation at 6 months (P < 0.05), but there was no statisticaly significant difference in the change rate of oxygen partial pressure and the gray value of lactate dehydrogenase between the fresh aminotic membrane and the cryopreserved aminotic membrane by vitreous cryopreservation at 3 months. The present study led to the conclusion that vitreous cryopreservation protocol alows to not only maintain the integrity of AM, but also to preserve the viability of the cels. So the vitreous cryopreservation is superior to the deep-frozen cryopreservation for cryopreservation of aminotic membrane.

Objective To evaluate the safety and effectiveness of the treatment by selective β 1 receptor blockers on patients with chronic obstructive pulmonary disease (COPD).Methods Eighty cases of COPD Ⅲ (stable period) inpatient with or without coronary heart disease were collected in The Second People's Hospital of Shanghai from September 2012 to November 2013.The patients were randomly divided into testing group (Metoprolol treatment group) and control group (regular treatment group) with 40 cases for each group.Metoprolol group therapy based on the use of conventional metoprolol tablets,an initial dose of metoprolol 12.5 mg/d,titrated to the appropriate dose based on heart rate and tolerance of the morning resting heart rate of 55 to 60 times/min that reached the target dose of metoprolol continuous medication for 12 months.Blood gas analysis were recorded before and after treatment,pulmonary function,and 6 min walk test (6MWT) and were chronic lung disease Assessment Test (CAT) Rating.The control group was administrated regular treatment while the testing group added small dose of Metoprolol with titration to an appropriate dose on this basis.12 months in a row,and assessed the end stage.Results (1) After the application of selective β receptor blockers on testing group,no statistically significant difference (P＞0.05) in the values of FEV1 in anticipation value％ (testing group:(45.45 ± 4.68) ％ vs.(43.32 ± 4.84) ％;control group:(44.23 ± 4.68) ％ vs.(42.58 ±4.24)％),PaO2(testing group:(75.92± 10.78) mmHg vs.(74.86± 11.21) mmHg;control group:(70.23 ±6.45) mmHg vs.(72.36±7.28) mmHg) and PaCO2(testing group:(46.28±8.28) mmHg vs.(47.46±10.22) mmHg);control group:(44.54 ± 8.89) mmHg vs.(42.36 ± 7.45) mmHg) before and after treatment.But the 6MWD (testing group:(287 ± 23) m vs.(384± 34) m;control group:(284 ± 25) m vs.(295 ±21) m) and COPD appraisal test(CAT) (testing group:(21±7) score vs.(17±6) score);control group:(22 ±5) score vs.(20± 6) score) had improved significantly compared with that before treatment,with significant difference(t=4.903,4.784;P＜0.05).Conclusion Selective β receptor blockers have no effect on the airway resistance of COPD patients and reduction on pulmonary function.It can also increase the exercise tolerance and enhance the living quality for improving clinical prognosis.

Objective To evaluate the outcomes of the payment reform at public hospitals in Sanming city.Methods Interrupted time series analysis was used to compare changes of the average days of stay,per capita hospitalization expense,outpatient expense per visit,proportion of medical expense and that of drugs during hospitalization at 21 public hospitals at or above county level before and after the DRGs reform.Results Comparisons before and after the reform found the average days of stay at the original momentum,poor control in curbing the proportion of medical expense and that of drugs during hospitalization,adropping followed by rising trend in the outpatient expense per visit,and minimal drop of the abovementioned proportions.Conclusions The rapid growth of outpatient and hospitalization costs at tertiary hospitals may be incurred by unreasonable cost transfer,structural trend of hospitalization expense makeup,and rationality pending scrutiny.

Tripartite-sector reform (a synergistic reform in public health services,medical insurance and medicine production-circulation) in Sanming city was described in the paper which centers on medical insurance.Tapping full potentials of the medical insurance,the city achieved efficient synergy among healthcare,medical insurance and medication systems.This reform has trimmed out inflated drug pricing to some extent for rooms of maneuver of medical service pricing changes,thus curbing excessive growth of medical costs successfully.The authors proposed areas of further improvements including the relationship between achieving such objective as curbing medical expenditure,and advancement of technical/medical service capacity;that between integrative control of medical insurance expenditure and protection of people's health;the equilibrium of interests between medical insurance,healthcare and medication.All these will contribute to the goal of healthy patients flow and a hierarchical medical system.

Objective To investigated to implement condition of preventive measure and control effect for endemic fluorosis in Fengshun County from 2005 to 2006.Methods It was investigated according to the National Surveillance Program of Endemic Fluorosis.Hupo,Daizai and Anquan Villages of Tangxi Town in Fengshun County were selected as monitoring spots.The usage of reforming water facilities,fluoride content in drinking water and urine of children aged 8-12 years and the prevalence rate of dental fluorosis of children were investigated.Resul tsin 2005 and 2006.a total of 18 reforming water facilities were surveyed and six of which were damaged or out of service.In 2005,the fluoride content in drinking water in the 3 villages was 2.10,1.22 and 0.15 mg/L The prevalence rate of dental fluorosis of children aged 8-12 years was 54.23%(64/118),38.91%(79/203) and 9.10%(6/66).The urine fluoride content of children was 0.95,0.90 and 1.05 mg/L,respectively.In 2006,the fluoride content in drinking water in Hupo,Daizai and Anquan Village was 2.01,1.57 and 0.21 mg/L.The prevalence rate of dental fluorosis of children aged 8-12 years was 26.47%(27/102),12.50%(23/184)and 6.15%(4/65),respectively.The urine fluoride content of children was 0.97,0.61 and 0.59 mg/L.Conclusions The outcome of surveillance data in Fengshun County has reached the sanle level as that of non-disease area.However,the management of reforming water facilities should be improved.

To explore the plasticity of human amniotic mesenchymal cells(hAMCs) into cardiomyocyte-like cells,hAMCs were isolated from human amnion with collagenase digestion.Phenotype of the isolated cells was analyzed by flow cytometry(FCM).hAMCs were treated with 5-azacytidine and basic fibroblast growth factor(bFGF) to investigate their ability of differentiation into cardiomyocytes.The induced differentiated cells were evaluated by immunofluorescence for desmin and ?-actin expression and by RT-PCR for Nkx2.5,GATA-4 and alpha-myosin heavy chain(?-MHC) mRNA expression.The results showed that,after primary culture,hAMCs could reach a confluence of 80% with swirl like growth at 6 days.The cells proliferated rapidly after passages with a 100% confluence at 3~4 d.hAMCs were positive expression of CD44 and vimentin,but negative for CK19.After induced differentiation at 8~10d,the differentiated cells have close-up arranged with long spindle-shape.At 2 weeks and 4 weeks,induced cells expressed ?-actinin and cardiac-specific transcription factor Nkx2.5.Expressions of GATA-4 and desmin can be detected but ?-MHC can not in the hAMCs both before and after the induction.In conclusion,hAMCs have the ability of differentiation into cardiomyocyte-like cells,which means that hAMCs can be regarded as candidate cells for cellular cardiomyoplasty(CCM).

Objective To analyze the association of miR-326 mRNA expression level on regulatory T cells between clinical manifestations of patients with systemic lupus erythematosus (SLE), in order to inves-tigate the role of miR-326 on Treg cells and pathogenesis as well as its association with disease activity of SLE. Methods Twenty milimeter anti-coagulated peripheral blood was obtained from 52 SLE patients and 20 healthy controls. Treg were purified by MACS from peripheral blood, in which the quantity of miR-326 and Ets-1 mRNA were assessed by real-time polymerase chain reaction (PCR). Data were analyzed by using Mann-Whitney U test and Spearman correlation analysis. Results Compared with healthy controls [0.921(0.345, 1.879)], the expression of miR-326 mRNA level was significantly higher in Treg from SLE patients [2.927 (0.518, 8.662) (Z=-3.756, P<0.05). The difference between new-onset SLE patients [8.878(5.922, 12.466)] and recurrence group [3.512(0.582, 11.223)] with healthy controls was significant (Z=-2.135, Z=-4.928, P<0.05). The expression level of miR-326 in new-onset SLE patients was higher than inactive SLE patients (Z=-4.928, P<0.05). Significant difference of the expression level of miR-326 mRNA was found between new-onset SLE patients with serous cavity effusion [7.606(0.656, 16.795)] and new-onset SLE patients without it [1.840(0.576, 13.025)](Z=4.263, P<0.05). Our analysis showed that significant positive correlation was found between the expression of miR-326 mRNA in Treg with CRP (rs=0.481, P<0.05) and anti-C1q antibody (rs=0.729, P<0.05) from new-onset SLE patients. Conclusion The expression level of miR-326 is upregulated in Treg from SLE patients and is associated with disease active index, which suggests that miR-326 in Treg may participate the pathological process and disease activity in SLE.

Objective: To investigate the clinical features and diagnostic bases of childhood leukoencephalopathy with cerebral calcifications and cysts (LCC). Methods: The clinical data involving manifestations and laboratory examinations of 4 children with LCC admitted to Beijing Children's Hospital Affiliated to Capital Medical University from 2012 to 2017 were retrospectively summarized. Each patient had a follow-up visit ranging from 4 months to 5 years and 9 months after initial examination. Results: Patients consisted of 2 males and 2 females, whose age of onset was respectively 2 years and 9 months, 6 years and 2 months, 7 years and 10 months, and 5 years and 1 month. The main clinical symptoms of these cases included headache, dizziness, partial seizure and claudication, and two of these cases had insidious onset. Cerebral calcifications and cysts with leukoencephalopathy were detected by neuroimaging in all patients. In addition, multifocal microhemorrhages and calcifications were observed by magnetic susceptibility-weighted imaging (SWI) series in 3 patients. Brain biopsy performed on 1 case disclosed a neuronal reduction in the cerebral cortex, loosening of focal white matter, multifocal lymphocyte infiltration, fresh hemorrhages, and gliosis, as well as angiomatous changes of blood vessels with hyalinized thicken-wall, stenotic or occlusive lumina and calcification deposits. The compound heterozygous mutations of n.*10G>A and n.82A>G in SNORD118 were identified in 1 case by target-capture next-generation sequencing. Sanger sequencing verified that the variant n.*10G>A was a novel mutation and it was of paternal-origin, while the variant n.82A>G was of maternal-origin, which had already been reported to be pathogenic to LCC. Follow-up study had shown continued partial seizure in 1 case and remissive claudication in another, while the remaining 2 cases had a relatively favorable outcome without obvious neurological symptoms at present time. Conclusions The clinical manifestations of LCC are nonspecific, and the onset of the disease tends to be insidious. The triad neuroimaging findings of cerebral calcifications, cysts and leukoencephalopathy are essential to the diagnosis of the disease, and the signals of microhemorrhages revealed by SWI series provide another eloquent reference for the diagnosis. As biopsy is invasive and usually unavailable in the early stage, gene assessment, instead of pathological data, should be the gold standard in the diagnosis of LCC.

Objective To compare target dosimetric distribution and normal tissue radiation between different static intensity?modulated radiation therapy ( IMRT) plans and volumetric modulated arc therapy ( VMAT) and to identify the best IMRT plan for patients with primary mediastinal B?cell lymphoma ( PMBCL) . Methods A total of 16 patients ( 8 males and 8 females) with early?stage ( Ann?Arbor stageⅠ) PMBCL were enrolled in this study,with doses of 45 Gy for primary gross tumor volume ( PGTV) and 40 Gy for planning target volume (PTV).Four plans were designed for each patient,consisting of static IMRT (5F?IMRT,7F?IMRT,9F?IMRT) and VMAT,and the target dosimetric distribution,normal tissue radiation dose,and efficiency of each plan were evaluated. The difference of dose was analyzed by analysis of variance. Results The mean conformity index ( CI) and homogeneity index ( HI) for PGTV in 5F?,7F?,9F?IMRT and VMAT were 1. 01 and 1. 10, 1. 01 and 1. 10, 1. 01 and 1. 10, and 1. 01 and 1. 11 ( P= 0. 963 and 0. 843) ,respectively,while these two indices for PTV were 1. 04 and 1. 22,1. 03 and 1. 19,1. 03 and 1. 17, and 1. 08 and 1. 14( P=0. 964 and 0. 969) ,respectively. The parameters of volume and dose were similar on normal tissue ( P= 0. 192?1. 000 ) . The treatment time and number of monitor units in 9F?IMRT were significantly higher than those in other static IMRT plans and VMAT ( P=0. 000,0. 000) ,and among these plans,VMAT had the lowest number of monitor units ( 13 345. 0 MU) and the shortest treatment time ( 5. 9 min) . Conclusions The target volume coverage of 7F?and 9F?IMRT is better than that of 5F?IMRT and VMAT.For early?stage PMBCL,VMAT is not superior to IMRT in terms of dosimetry,especially with a larger area of low?dose radiation to the breast,but it is highly efficient in practice.

Objective To prospectively evaluate the efficacy and toxicity of hypofractionated intensity-modulated radiotherapy (IMRT) for prostate cancer.MethodsFifty-two consecutive patients with localized prostate cancer were enrolled in this study between Feb.2009 and Mar.2011.All patients received hypofractionated IMRT (2.7 Gy/fx,25 fractions,total 67.5 Gy) to the prostate and seminal vesicles.32 high risk patients also received prophylactic irradiation to the pelvic lymph nodes concurrently (2 Gy/fx,25 fractions).Imaging-guided radiotherapy was employed in 35 patients.Androgen deprivation therapy was adopted in 48 of 52 patients.ResultsAfter a median follow-up of 13 months,the mean prostate specific antigen (PSA) was reduced from (40.3 ± 36.6) ng/ml before treatment to (0.5 ± 1.7)ng/ml at the last follow-up.By the time of last follow-up,2 patients (4％) failed.One had PSA failure and the other had both PSA failure and pelvic lymph node relapse.25％ of the patients experienced grade 2 acute gastrointestinal (GI) toxicity and 4％ experienced grade 3 GI toxicity.Acute grade 2 and grade 3genitourinary ( GU ) toxicity occurred in 15％ and 2％,respectively.The incidence of late grade 2 and grade 3 GI toxicity was 17％ and 0％,respectively.Late grade 2 and 3 GU toxicity was 8％ and 2％.The potency was unable to evaluate because most of the patients received androgen deprivation therapy.Conclusions The short-term PSA-free survival after 2.7 Gy/fx,25 fractions' hypofractionated IMRT for localized prostate cancer is favorable,and the acute and late GI and GU toxicity are acceptable.A longer time follow-up is warranted to ascertain the long term efficacy and safety of this regimen.