Morphometry of the ureteric bud branching in the developing mouse kidney / 解剖学报

[Abstract] Objective To investigate the branching pattern of the ureteric bud and the number of the nephron induced by each ureteric bud tip, through the three-dimensional tracing of the ureteric tree, combined with the morphological analysis and measurement of the ureteric tree. Methods The kidneys were obtained from three mice at various developing time points and prepared for paraffin and epoxy sections. Then the microscopic images were digitized and aligned from these sections. Based on the computer-assisted tracing and visualization of ureteric tree, the number of branches and the nephron induced by each ureteric bud tip were obtained by counting. In addition, paraffin sections were stained with HE staining for morphological observation of nephrogenic zone and ureteric bud, while in order to reflect the density of the ureteric bud tips at nephrogenic zone, the distance between two neighboring ureteric bud tips was measured aided with the Claudin-7 immunohistochemical staining. Results The ureteric bud branching tree revealed that the initial bifid iterative branching formed the framework of renal medulla, the branching became complicated and dense in cortex and nephrogenic zone, while the distance between ureteric bud tips were also decreasing. The number of the nephron induced by each ureteric bud tip increased from one (E14. 5) to two (E17. 5), and occasionally to three. Conclusion Threedimeasional Visualization of ureteric bud branching tree reveals regional complication, suggesting molecules in different regions drive different branching patterns; While the density of the ureteric bud tips at nephrogenic zone increases corresponding to decreasing of thickness of the nephrogenic zone, and the disappearance of the ureteric bud tips after birth is also consistent with the gradual consumption of nephron progenitor cells.

Three-dimensional visualization of the renal proximal tubules in developing mice / 解剖学报

Objective Adult proximal tubule ( PT ) is not only the segment most frequently involved in acute renal tubule injury, but also the easiest to repair. It may be consistent with the rapid growth and differentiation mechanism of this segment during the development of the kidney, while the developing information is insufficient. Therefore, we three- dimensional visualized the developing PT to analysis its spatiotemporal morphogenesis. Methods The kidneys were obtained from mice at various developing time point, embryonic day ( E ), postnatal day ( P ). The volume density of Claudin-2 positive PT in the cortex was measured using a stereological method in paraffin sections. After image recording and alignment of the serial sections, the spatial courses of the developing PT were traced and visualized in three dimensions using computer-assisted program. The length of the developing PT was calculated at the same time. Results The volume density of PT in the cortex of PI mice was significantly higher than that in the embryonic stage. Then it experienced a decline ( P3, P5 ), an increase ( start at P7 ) to a stable adult level ( P28 ). The tubular tracing showed that the lengths of developing PT and the number of convolutions of their convoluted part increased with the maturation, but lower than that of adultin E14. 5, E17. 5 and P5 PT in E14. 5 and E17. 5 mice were similar to that of adult with respect to general spatial courses. They were, however, significantly different from adult in the initial direction of PT and the arrangement of the straight part of PT in the medullary rays. While, it was in P5 that the spatial pattern of some PT was gradually approaching to the adult model. Conclusion This study demonstrated that the development of PT was consistent with the kidney development in terms of its volume density in cortex, length and spatial course. It started at the S-shaped body, kept throughout the embryonic period and continued to postnatal, ended at kidney maturation ( P28 ).

Variant of Associated with Increasing Risk in Chinese Patients with Relapsing-remitting Multiple Sclerosis / 中华医学杂志(英文版)

<p><b>Background</b>Multiple sclerosis (MS) is a common central nervous system autoimmune disorder. Increasing number of genome-wide association study (GWAS) analyses hint that MS is strongly associated with genetics. Unfortunately, almost all the GWAS analyses were Caucasian population based. Numbers of risk loci might not be replicated in Chinese MS patients. Hence, we performed a MassArray Assay to genotype the previously reported variants located in the transcription regulation genes in order to elucidate their role in the Chinese MS patients.</p><p><b>Methods</b>One hundred and forty-two relapsing-remitting MS (RRMS) patients and 301 healthy controls were consecutively collected from September 2, 2008, to June 7, 2013, as stage 1 subjects. Eight reported transcription regulation-related single-nucleotide polymorphisms (SNPs) were genotyped using the Sequenom MassArray system. In stage 2, another 44 RRMS patients and 200 healthy controls were consecutively collected and Sanger sequenced from April 7, 2015, to June 29, 2017, for the validation of positive results in stage 1. Differences in allele and genotype frequencies between patients and healthy controls, odds ratios, and 95% confidence intervals were calculated with the Chi-square test or Fisher's exact test. Hardy-Weinberg equilibrium was tested also using the Chi-square test.</p><p><b>Results</b>In stage 1 analysis, we confirmed only one previously reported risk variant, rs11129295 in EOMES gene. We found that the frequency of T/T genotype was much higher in MS group (χ = 10.251, P = 0.005) and the T allele of rs11129295 increased the risk of MS (χ = 10.022, P = 0.002). In stage 2 and combined analyses, the T allele of rs11129295 still increased the risk of MS (χ = 4.586, P = 0.030 and χ = 16.378, P = 5.19 × 10, respectively).</p><p><b>Conclusions</b>This study enhances the knowledge that the variant of EOMES is associated with increasing risk in Chinese RRMS patients and provides a potential therapeutic target in RRMS.</p>

Safflower Injection combined with alprostadil and sildenafil treats the chronic pulmonary heart disease complicated with pulmonary hypertension / 中成药

AIM To observe the effect of Safflower Injection (safflower yellow) combined with alprostadil and sildenafil in the treatment of chronic pulmonary heart disease (chronic cor pulmonale) complicated with pulmonary hypertension.METHODS Two hundred and twenty-three cases of chronic pulmonary heart disease complicated with pulmonary hypertension patients were randomly divided into two groups,the control group of one hundred and eleven cases in conventional therapy plus alprostadil and sildenafil,one hundred and twelve cases in the treatment group were treated with Safflower Injection on the basis of the control group.The efficacy and side effects in two groups were observed.RESULTS The total effective rate of the treatment group was significantly higher than that of the control group.Pulmonary artery systolic pressure (SPAP),pulmonary arterial mean pressure (mPAP) and pulmonary artery diastolic blood pressure (DPAP) in two groups,compared with those before the treatment,were significantly decreased,left ventricular shoot ejection fraction (LVEF),compared with those before treatment,was significantly increased;SPAP,mPAP and DPAP of treatment group after the treatment were decreased significantly as compared with the control group,LVEF was significantly increased.Arterial partial pressure of oxygen (PaO2) and arterial oxygen saturation (SaO2) in two groups,compared with those before the treatment,were significantly increased,carbon dioxide partial pressure (PaCO2),compared with those before the treatment,was significantly decreased;after treatment in the treatment group,PaO2 was significantly increased as compared with the control group.Blood high shear viscosity,whole blood viscosity at low shear,plasma viscosity,red blood cell pressure volume and platelet aggregation rate in two groups were significantly lower than those before the treatment;whole blood high shear viscosity,whole blood viscosity of low shear,plasma viscosity and platelet aggregation rate in the treatment group,compared with the control group,were decreased significantly.High sensitive C reactive protein (hsCRP),creatine kinase (CK-MB) and cardiac troponin Ⅰ (TnⅠ) in two groups,compared with those before the treatment,were significantly reduced;after the treatment,hs-CRP,CK-MB and TnⅠ in the treatment group,compared with the control group,were decreased significantly.There was no difference in incidence of adverse reactions between the two groups.There were no significant changes in liver and renal functions before and after the treatment in two groups.CONCLUSION Safflower Injection combined with alprostadil and sildenafil in the treatment of chronic pulmonary heart disease complicated with pulmonary hypertension has a good curative effect.

Variants of Interleukin-7/Interleukin-7 Receptor Alpha are Associated with Both Neuromyelitis Optica and Multiple Sclerosis Among Chinese Han Population in Southeastern China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Neuromyelitis optica (NMO) and multiple sclerosis (MS) are autoimmune demyelinating diseases of the central nerve system. Interleukin-7 (IL-7) and interleukin-7 receptor alpha (IL-7Rα) were proved to be important in the pathogenesis of both diseases because of the roles they played in the differentiations of autoimmune lymphocytes. The variants of both genes had been identified to be associated with MS susceptibility in Caucasian, Japanese and Korean populations. However, the association of these variants with NMO and MS has not been well studied in Chinese Southeastern Han population. Here, we aimed to evaluate the association of six IL-7 variants (rs1520333, rs1545298, rs4739140, rs6993386, rs7816065, and rs2887502) and one variant of IL-7RA (rs6897932) with NMO and MS among Chinese Han population in southeastern China.</p><p><b>METHODS</b>Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MassARRAY system) and Sanger sequencing were used to determine the variants of IL-7 and IL-7RA in 167 NMO patients, 159 MS patients and 479 healthy controls among Chinese Han population in southeastern China. Samples were excluded if the genotyping success rate <90%.</p><p><b>RESULTS</b>Statistical differences were observed in the genotypes of IL-7 rs1520333 in MS patients and IL-7RA rs6897932 in NMO patients, compared with healthy controls (P = 0.035 and 0.034, respectively). There was a statistically significant difference in the genotypes of IL-7 rs2887502 between MS and NMO patients (P = 0.014). And there were statistically significant differences in the rs6897932 genotypes (P = 0.004) and alleles (P = 0.042) between NMO-IgG positive patients and healthy controls.</p><p><b>CONCLUSIONS</b>The study suggested that among Chinese Han population in southeastern China, the variant of IL-7RA (rs6897932) was associated with NMO especially NMO-IgG positive patients while the variant of IL-7 (rs1520333) with MS patients. And the genotypic differences of IL-7 rs2887502 between MS and NMO indicated the different genetic backgrounds of these two diseases.</p>

The mediating role of parenting in relationship between parent hostility and adolescent externalizing behavior problems / 中华行为医学与脑科学杂志

Objective To explore the relationship between parent hostility and adolescent externalizing behavior problems and the mediating role of parenting.Methods The cross-sectional data of 4474 adolescent aged 10-21 of the National Survey of Adolescent Mental Health in China (2008) was used.Adolescents reported their externalizing problems and parents' parenting.Parents reported their hostile behaviors.Results ① Parent hostility((9.18 ± 3.56),(8.94 ± 3.18)) significantly correlated with monitoring,inductive reasoning,harshness of parenting(father(3.39 ± 1.16),(3.21 ± 1.02),(1.39 ± 0.62) respectively,r =-0.108 ～ 0.489,P ＜ 0.01 ;mother (3.96 ± 0.97),(3.38 ± 0.96),(1.37 ± 0.60) respectively,r =-0.148 ～ 0.468,P ＜ 0.01) ; and both parent hostility and parenting were significantly associated with adolescent externalizing problems (0.23 ± 0.21) (r =-0.308 ～ 0.577,P ＜ 0.01).②Parental monitoring,inductive reasoning and harshness discipline partially mediated the relations between parent hostility and adolescent externalizing problems (Z =6.12,5.10,6.36,P ＜ 0.01 ; Z =5.86,6.97,11.84,P ＜ 0.01).Conclusion Parenting partially mediated the effect between parent hostility and adolescent externalizing problems.

Effects of a pulsed electromagnetic field on cell proliferation, differentiation and osterix mRNA expression in rat osteoblasts / 中华物理医学与康复杂志

Objective To investigate the effects of a pulsed electromagnetic field (PEMF) on cell proliferation,differentiation and the genetic expression of osteogenesis specific transcription factor-Osterix (OSX) in the calvaria-derived osteoblasts of SD rats.MethodsPrimary osteoblasts were separated from the calvarias of Sprague-Dawley rats 24 hours after birth by trypsinazation and collagenase digestion.The osteoblasts at passage 4 were randomly divided into a control group and a PEMF group,Cells in the PEMF group were placed onto PEMF therapeutic apparatus and exposed to 11 mT radiation at 12 Hz for 1 h per day for 3 days.The osteoblasts' proliferation ability was then detected via methylthiazdyl tetrazolium assay.Alkaline phosphatase (ALP) activity in the cell lysate and the supernatant fluid was assayed through disodium phenyl phosphate colorimetric determination to determine the cells'differentiation ability.OSX mRNA expression was determined using real time reverse transcription polymerase chain reaction (RT-PCR).ResultsThe proliferation of osteoblasts in the PEMF group was significantly greater than in the control group.PEMF exposure suppressed ALP activity in both the lysate and the supernatant of the osteoblasts.The OSX mRNA expression in the PEMF group was significantly down-regulated compared with the control group.ConclusionPEMF at 12 Hz and 11 mT can promote the proliferation of osteoblasts,but it inhibits cellular differentiation and down-regulates the mRNA expression of OSX.

Controlling effect of bushen huatan compound on the insulin signal conducting molecule inside ovaries in polycystic ovary syndrome model rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of Bushen Huatan Compound (BHC) on the glycolipid metabolism and the expressions of the insulin signal conducting molecules inside ovaries in polycystic ovary syndrome (PCOS) model rats.</p><p><b>METHODS</b>Female Wistar rats were subcutaneously injected with 2.5 mg/kg testosterone propionate (Their female offspring were randomly divided into the medication group and the model group, 10 in each.) or neutral tea oil of the same dose (Ten female offspring was taken as the control group.) on the 16th day of pregnancy, once daily, for 3 successive days. BHC was given to rats in the medication group by gastrogavage, while equal volume of distilled water was given to rats in the model group and the control group by gastrogavage, both once daily for 20 successive days. The body weight and ovary weight were weighed to calculate the ratio of wet fat weight/body weight. The blood glucose levels were detected at 0, 0.5, 1, and 2 h using oral glucose tolerance test (OGTT). The serum concentrations of high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), fasting blood glucose (FBG), and insulin were detected to calculate homeostasis model assessment of insulin resistance (HOMA-IR). The expressions of protein kinase B (AKT2), glycogen synthase kinase-3beta (GSK3beta), glucose transporter-4 (GLUT4), extracellular signal regulated kinase-1 (ERK1) protein, P-AKT2, P-GSK3beta, and P-ERK1 in ovaries were detected using Western blot.</p><p><b>RESULTS</b>Compared with the control group, the ratio of wet fat weight/ body weight, the blood glucose levels at 0.5 and 2 h in OGTT, and HOMA-IR all obviously increased, and the HDL-C level obviously decreased in the model group (P < 0.05). Compared with the model group, the ratio of wet fat weight/body weight and the blood glucose levels at 2 h in OGTT obviously decreased, and the HDL-C level obviously increased in the medication group (P < 0.05). The expressions of AKT2, P-AKT2, GSK3beta, P-GSK3beta, GLUT4, and ERK1 in the ovary tissue were obviously lower in the model group than in the control group (P < 0.05). The expressions of GSK3beta, P-GSK3beta, and GLUT4 were more obviously enhanced in the medication group than in the model group (P < 0.05).</p><p><b>CONCLUSIONS</b>Insulin resistance and glucolipid metabolism dysfunction existed in female PCOS rats. Besides, abnormal insulin signaling pathway existed in the ovary tissue. BHC could remarkably ameliorate the IR degree and glucolipid metabolism functions, and might be correlated with regulating the protein expressions of insulin signal conducting molecules.</p>

Patient safety monitoring indicators based on medical complaints / 中华医院管理杂志

Objective To build the indicators system to collect patient safety monitoring information, focusing on medical complaints. Methods With such methods as literature review and expert advice, building the system for medical complaints collection and monitoring. Such indicators are modified and improved in pilot operations. Results The framework of the medical complaint monitoring indicators system is built in five dimensions, comprising 8 grade-1 indicators including patient complaint causes and hospital cause analysis, and 20 grade-2 indicators. Conclusion These indicators are scientific and operable to detect adverse patient safety events.

Regulation of synthesis function of porcine ovarian granulose cells by mitogen-activated protein kinase signaling pathway / 中华妇产科杂志

Objective To investigate the role of mitogen-activated protein kinase(MAPK)signaling pathway regulating synthesis function of steroid hormone of porcine ovarian granulose cells.Methods Porcine ovarian granulose cells were cultured in medium added with dimethyl sulfoxide(DMSO)at concentration of 50μmol/L or with MAPK inhibitor PD98059 (PD) for 48 hours.Then,granulose cells in DMSO medium were added with activator of adenylate cyclase(20 μmoL/L)or blank agent for next 48 hours incubation,which were defined as observation group 1 and control group 1,similarly,granulose cells in PD medium were also added with activator of adenylate cyclase(20 μmol/L)or blank agent for next 48 hours,which were defined as observation group 2 and control group 2.The level of estradiol(E_2),progesterone (P)and testosterone (T) were detected by chemoluminescence.The mRNA expression of 17alpha-hydroxylase/17,20-lyase (CYP17) and aromatase P450 (P450 arom) were assessed by RT-PCR.Results (1)Hormone expression:the level of T,P and E_2 were(29.5±2.5)nmol/L,(80±5)nmol/L,(49±4) pmol/L in control group 1 and(42.3±3.4)nmol/L,(170±15)nmol/L,(75±6)pmol/L in control group 2,which showed significant difference(P<0.05).In the mean time,the level of T, P and E_2 were (106.2±7.6)nmol/L,(210±16)nmol/L,(130±11)pmol/L in observation group 2 and(47.2±3.5)nmol/L,(130±6)nmol/L,(81±6)pmol/L in observation group 1,which also reach statistical difference(P<0.05).(2)The expression of enzyme:the expression of CYP17 mRNA was inereased by 50% and P450 arom mRNA was decreased by 20% between control group 1 and 2. However, the mRNA expression of P450 arom and CYPI7 were upregulated remarkably, especially, the expression of CYP17 mRNA were increased by 125%. Conclusion MAPK signaling pathway plays an inhibitory role in regulating synthesis of steroid hormone of ovarian granulose cell.

Reform and Pondering of Theory Exam on Histo-Embryology / 中华医学教育探索杂志

The exam under open and close condition is a new mode which is different from traditional close-exam and entire open-exam.It indicates that new exam mode not only conquers limitation of simplex close-exam,but also boosts forwardly study of students and promotes the didactical reform completely.

Preparation of xenogenic corneal stroma and its cytoconsistency / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the possibility that using the bovine corneal stroma to provide a suitable carrier on which the cells can grow for tissue engineering cornea.</p><p><b>METHODS</b>Nine fresh bovine corneas were selected. Each cornea was cut into 2 pieces, and exposed to 0.25% trypsinase for various lengths of time (20 minutes, 40 minutes, and 60 minutes) to get the stroma part with least cells and maintaining the collagen fibers arrangement. Samples obtained from each group were examined with scanning electron microscopy and HE staining. The left ones were freeze-dried and sterilized. The various concentrations of extraction were used to cultivate human fibroblasts, and a 3-(4,5-dimethylthiazol-2-yl)-2, (MTT)-based colorimetric assay was taken to evaluate the exhistance of 5-diphenyltetrazolium bromide cytotoxinic effects. Then the proper corneal stroma was used as a carrier to cultivated the rabbit corneal limbal cells which were planted on it in a concentration of 2 x 10(5)/cm2 in vitro. The cell-carrier samples were sent for scanning electron microscopy and HE staining.</p><p><b>RESULTS</b>The corneal stroma had the least cells in the group acted with typsin for 60 minutes, while the collagen fibers arrangement was not so orderly as before. The extractions showed no significant difference in cell culture, and no obviously harmful effect on the cell growth. The rabbit corneal limbal cells presented a stratified growth on the bovine corneal stroma.</p><p><b>CONCLUSION</b>The bovine corneal stroma without cells prepared using the typsin and lyophilization can be a suitable carrier for cell culture in vitro.</p>