This paper reviews and summarizes the background，stages，tasks，achievements and experiences of the China's new round of healthcare reforms. Lessons gained from the reforms are valuable for deepening the healthcare reforms of China.

Objective To analyse MMACHC mutations for 45 pedigrees with combined methylmalonic aciduria and homocyctinuria by Sanger sequencing, and to discuss the utility of prenatal genetic diagnosis for these pedigrees.Method Peripheral blood was collected from 45 probands and their parents from 2012-2015 in Genetic Counselling Clinic of the First Affiliated Hospital of Zhengzhou University, and the DNA were extracted from the blood.Then the coding sequence of MMACHC gene was amplified by PCR, and the PCR products were further sequenced to detect mutations for each pedigree.For 12 families, chorionic villus sampling was performed on the pregnant women to make prenatal genetic diagnosis.Result There were 14 distinct mutations detected in the 45 pedigrees, and the most frequent mutations are c.609G>A(W203X),c.658-660delAAG(K220del)and c.80A>G (Q27A).Two of those mutations have not been reported before:one is a splicing site mutation c.81+1G>A;while the other is a missense mutation c.665A>G,p.Y222C.Most mutations were found in exon 4.Among the 12 pedigrees who received prenatal diagnosis, 2 fetuses were normal, 7 fetuses were carriers of heterozygous mutation, and the other 3 fetuses were patients with compound heterozygous mutation or homozygous mutation.The couples whose fetuses were normal or carriers continued the gestation, while the couples whose fetuses were patients decided to terminate the pregnancy.After delivery, the outcome of the fetuses was the same as the prenatal diagnose results.Conclusion Two novel mutations of MMACHC were identified and prenatal genetic diagnosis helps to avoid the delivery of combined methylmalonic aciduria and homocyctinuria patients.

Vascular endothelial growth factor (VEGF) plays an important role in tissues angiogenesis. The adaptation of animals to hypoxic environment is relative to the microvessel density (MVD) in tissues. To further explore the adaptation mechanisms of plateau zokor (Myospalax baileyi) to the hypoxic-hypercapnic burrows, the VEGF mRNA and the MVD in cerebral tissues of the plateau zokor were studied. Total RNA was isolated from liver, and VEGF cDNA was obtained by RT-PCR, then the VEGF cDNA was cloned and sequenced. The coding sequence of plateau pika (Ochotona curzniae), rat (Rattus norvegicus) and mouse (Mus musculus) VEGF cDNA are obtained from GenBank, and the nucleotide and amino acid sequence homology of plateau zokor VEGF cDNA coding sequence with that of plateau pika, rat and mouse were analyzed and compared by using of bioinformatics software. The VEGF mRNA was detected by real-time PCR, and the MVDs in cerebral tissues of the plateau zokor, plateau pika and Sprague-Dawley (SD) rat were measured by immunohistochemical staining. The results showed that the open reading frame of the plateau zokor VEGF was 645 bp, and the coding sequence of the plateau zokor VEGF cDNA shared 92.1%, 93.6% and 93.8% nucleotide sequence homology to that of the plateau pika, rat and mouse, respectively. The deduced amino acid sequence of the plateau zokor VEGF cDNA was composed of 188 amino acids and the amino acids from 1 to 26 were signal peptide sequence. The plateau zokor VEGF188 was 90.2%, 94.9% and 94.4% homologous to that of plateau pika, rat and mouse. The level of VEGF mRNA in brain of the plateau zokor was significantly lower than that of SD rat, but there was no obvious difference in VEGF mRNA level between plateau zokor and plateau pika. The MVD in brain of the plateau zokor was markedly higher than that of plateau pika and SD rat. In conclusion, plateau zokor enhances its adaptation to the hypoxic environment by increasing the MVD. The level of VEGF mRNA in the brain of plateau zokor is lower than that of SD rat, which may be as a result of inhibition by the higher concentration of carbon dioxide in the burrow.
Adaptation, Physiological ; physiology ; Amino Acid Sequence ; Animals ; Arvicolinae ; physiology ; Base Sequence ; Brain ; blood supply ; metabolism ; Hypoxia ; physiopathology ; Microvessels ; anatomy & histology ; Molecular Sequence Data ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Species Specificity ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Current study was carried out to optimize the priming condition of Oldenlandia diffusa seeds, and improve germination rate and seed vigor of 0. diffusa seeds under drought conditions. Uniform design was used to optimize the concentration and priming time of three priming materials (PEG, KNO3, GA3). Different concentrations of polyethylene glycol (PEG) was used to simulate drought stress. The seedling was cultured in 1/4 Hoagland medium for 30 d. The results showed that seed priming treatment with 366 mg x kg(-1) GA3 for 1h resulted in significant increase in germination rate, germination index, vigor, root length, plant height and biomass of O. diffusa seeds under drought stress (15% PEG), while seed priming with 3.0% KNO3 for 1 h showed little effect on germination and growth of O. diffusa seeds under drought stress. Seed priming treatment with appropriate GA3 concentration and priming time could enhance seed germination and drought resistance of O. diffusa in seedling stage.
Droughts ; Germination ; Oldenlandia ; growth & development ; physiology ; Seedlings ; growth & development ; physiology ; Seeds ; growth & development ; physiology ; Stress, Physiological

OBJECTIVETo investigate the differences in semen quality between samples collected by masturbation in the clinic and at home.

METHODSBased on the WHO guidelines, we analyzed the ejaculates collected by masturbation in the clinic and at home from 342 men under infertility assessment and measured the contents of such biochemical markers in the seminal plasma as neutral α-glucosidase, zinc, and fructose. According to the location of semen collection, we divided the samples into two groups, clinic-collected and home-collected, and analyzed the differences in the semen parameters between the two groups with the SPSS 16.0 software.

RESULTSCompared with the clinic-collected semen, the home-collected samples had significantly higher mean values in semen volume (4.0 vs 4.9%), sperm concentration (41 vs 64 x 10(6)/ml), total sperm count (175 vs 270 x 10(6) per ejaculate), progressive sperm motility (40 vs 52%), total count of progressively motile sperm (82 vs 135 x 10(6) per ejaculate) (all P <0.05). No significant differences were found between the two groups in normal sperm morphology (4.0 vs 5.0%) and the contents of neutral α-glucosidase (26 vs 24 mU per ejaculate), zinc (8.0 vs 8.0 μmol per ejaculate), and fructose (62 vs 60 μmol per ejaculate) (all P >0.05). Abnormal sperm concentration (<20 x 10(6)/ml) was observed in significantly fewer of the home-collected samples than the clinic-collected ones (18% [62/342] vs 30% [103/342], P<0.05), and so was abnormal progressive sperm motility (<32%) (64% [219/342] vs 75% [256/342], P<0.05).

CONCLUSIONOur findings show that semen samples collected by masturbation at home has a higher quality than those collected in the clinic. So the location of semen collection should be taken into consideration in infertility investigation.

Humans ; Infertility, Male ; diagnosis ; Male ; Masturbation ; Semen ; enzymology ; physiology ; Semen Analysis ; methods ; Specimen Handling ; methods ; Sperm Count ; Sperm Motility ; alpha-Glucosidases ; analysis

OBJECTIVETo investigate the expressions and action mechanisms of nerve growth factor (NGF) receptors TrkA and p75NTR in the oncogenesis and progression of prostate cancer (PCa).

METHODSUsing immunohistochemistry, we detected the expressions of TrkA and p75NTR in 62 PCa and 35 benign prostatic hyperplasia (BPH) samples, and conducted statistical analysis on the basis of clinical data.

RESULTSIndependent-samples t-test showed that, along with poorer tissue differentiation or higher clinical stage of PCa, the expression of TrkA was significantly up-regulated, that of p75NTR remarkably down-regulated, and the expression ratio of TrkA to p75NTR markedly increased. The TrkA/p75NTR ratio was 0.32 in the BPH, 0.52 in the PCa tissue with Gleason score of 6, 1.65 in the PCa tissue with Gleason score of 7, 5.75 in the PCa tissue with Gleason score ≥ 8, 0.89 in the clinical stage of pT2, 1.5 in pT3 a, 3.75 in pT3b, and 7.00 in pTxN1.

CONCLUSIONThe abnormally increased expression ratio of TrkA to p75NTR might be one of the essential features of malignant transformation of prostate cells. A higher TrkA/p75NTR expression ratio may be associated with a lower tissue differentiation, a higher clinical stage or Gleason score, and therefore a poorer prognosis.

Humans ; Immunohistochemistry ; Male ; Neoplasm Grading ; Neoplasm Staging ; Nerve Tissue Proteins ; metabolism ; Prognosis ; Prostatic Hyperplasia ; pathology ; Prostatic Neoplasms ; pathology ; Receptor, trkA ; metabolism ; Receptors, Nerve Growth Factor ; metabolism ; Up-Regulation

Background Retinitis pigmentosa (RP)is a common hereditary blinding eye disease in ophthalmology.Current researches documented that RP may have the common pathophysiologic basis to Alzheimer disease and chronic neurodegenerative disease.Understanding this mechanism will offer a new therapeutic target for RP.Objective The purpose of the present study was to investigate the roles of cyclin-dependent kinase 5 (Cdk5)/P25 activation in the apoptosis of retinal neural cells of RCS rats.Methods Eighteen SPF RCS rats and 18 RCS-rdy+ rats were randomized into 17-,25-and 35-day groups respectively and 6 rats for each.The rats were sacrificed at corresponding time points and retinal hemogenete was prepared.Expressions of CdkS,P35,P25 and tau phosphorylation in the retinas were detected by Western blot,and the kinase activity of Cdk5/P25 was analyzed by quantitative colorimetric assay.Results The expressing level of P35 protein(A340) in the retinas of 17-day-old RCS rats was near that of 17-day-old RCS-rdy+ rats(t =0.52,P＞0.05).In 25-and 35-day-old RCS rats,the expressing levels of P35 protein were 2.20±0.48 and 1.23±0.14,which were higher than those of RCS-rdy+ rats(1.43±0.13 and 0.93±0.10),showing significant differences between them(t =3.78,4.28,P＜0.05).The expression of P25 was undetectable at postnatal 17 days in RCS rats and RCS-rdy+ rats,but it showed significantly higher in RCS rats(0.300±0.003 and 0.230±0.004) than that in RCS-rdy+ rats(0.040±0.004 and 0.070±0.004) at postnatal 25 days and 35 days(t=121.81,77.51,P＜0.01).No significant difference was found in the expression of Cdk5 in RCS rats and RCS-rdy+ rats at different ages (t =-0.60,0.19,1.62,P＞ 0.05).The kinase activity of Cdk5/P25 did not show significantly different between RCS and RCS-rdy+ rats at postnatal 17 days(t =0.19,P＞0.05),but significantly higher kinase activity of Cdk5/P25 was seen in RCS rats (0.0058 ±0.0005 and 0.0056±0.0004) than that in RCS-rdy+ rats(0.0038±0.0003 and 0.0032 ±0.0007) at postnatal 25 days and 35 days (t =8.07,5.97,P＜ 0.01).No expression of tau phosphorylation was detected in RCS rats at postnatal 17 days,but significantly higher tau phosphorylation level was seen in RCS rats at postnatal 25 days and 35 days(1.80±0.22 and 1.23±0.17),which were significant different in comparison with RCS-rdy+ rats at postnatal 25 days and 35 days(1.60 ±0.20 and 1.04 ±0.12)(t=4.71,3.17,P＜0.05).Conclusions The Cdk5/P25 kinase activity shows a consistent trend with theexpressions of P25 and tau phosphorylation in the RCS rats,indicating that the upregulation of P25 induces the enhance of enzyme activity of Cdk5,which phosphorylate its substrates to result in more apoptosis of retinal neural cells.

Objective To research the curative effect of chondroitin sulfate and glucosamine on Kashin-Beck disease(KBD). Methods According to Diagnosis for Kashin-Beck disease,80 patients of adult KBD were detected from Guanghui village Shangzhi city Heilongjiang province in July,2007,and they divided into treatment group and control group according to their condition,age and sex,40 person in each group. Treatment group was given chondroitin sulfate and glucosamine sulfate,and control group was given placebo(equivalent amount of starch). Bilateral knees X-ray films were shot before and after treatment (8th month),scale division magnifying glass was used to measure the width of joint space on X-ray films. Results The width of joint space respectively was (4.30±2.14) and (4.10±2.07)mm in control group before and after treatment,and treatment group respectively was (4.17±2.15),(4.16±2.11)mm. Medicine had no obviously role on joint space (F = 0.50,P > 0.05),Time and both of time and medicine had obvious role on joint space(F= 67.66,46.74,all P< 0.05). Joint space of treat group was thinner than control group(P < 0.05) before treatment,but thicker after treatment(P < 0.05). To compare with the width of before treatment,joint space width of control group became obviously narrow(P < 0.05). Conclusions Experimental group taking chondroitin sulfate and glucosamine sulfate alleviated knee joint space narrowing process of adults KBD patients compared with control group. It plays a protection role in articular cartilage and provides evidences for choosing drug and evaluating effect in the treatment of adults KBD.

OBJECTIVE: To access application value of multi-slice spiral computed tomography (MSCT) and coronary artery calcium scoring (CACS) in investigation the coronary artery disease (CAD), and to explore the effective way of virtual autopsy to evaluate the sudden death due to CAD. METHODS: Nine cases of sudden cardiac death were collected to analyze MSCT before the autopsy. The quantitative analysis of the degree of coronary artery calcium was made by Agatston's method. The CACS of all the subjects were calculated based on the diagnostic criteria for CAD, in which calcium scoring was more than 400. The results of CACS were compared with that of the autopsy. RESULTS: Only 2 cases got the high calcium scoring which were more than 400 in the 9 cases died of CAD confirmed by the autopsy. The prediction rate of CACS for CAD was only 22.2%. Pulmonary edema of different severity was found in both autopsy and MSCT. There was a higher morbidity rate in the left anterior descending of coronary artery than the other branches. CONCLUSION Obvious calcification of coronary artery can be detected by MSCT and calculating CACS. To detect subtle calcification needs other technologies such as postmortem angiography.
Autopsy ; Coronary Angiography ; Coronary Artery Disease/diagnostic imaging* ; Death, Sudden/pathology* ; Humans ; Multidetector Computed Tomography/methods* ; Predictive Value of Tests ; Vascular Calcification/diagnostic imaging*

OBJECTIVETo compare the efficacy of decollement, combined tumescent liposuction and curettage, Botox injection in the treatment of osmidrosis.

METHODSFrom September 2006 to April 2009, 350 patients with osmidrosis were treated by decollement method in 180 cases, curettage in 120 cases,and Botox injection in 50 cases. The therapeutic effect and complications were retrospectively analyzed.

RESULTSThe patients were followed up for 3-24 months. The cure rate was 90.6%, 84.8% and 84.2% in decollement, curettage and Botox groups, respectively. The effective rate was 100% in all groups.

CONCLUSIONSAll the three methods are effective for the treatment of osmidrosis. The odor symptom can be improved a lot with decollement method. Considering the complication, downtime and cosmetic results, Botox injection is suitable for patients with minor or median odor, while combined liposuction and curettage is selected for patients with median to severe odor.

Axilla ; surgery ; Botulinum Toxins, Type A ; administration & dosage ; therapeutic use ; Curettage ; methods ; Female ; Follow-Up Studies ; Humans ; Hyperhidrosis ; surgery ; Lipectomy ; methods ; Male ; Retrospective Studies ; Treatment Outcome