Objective To develop and compare the methods for determining the carrier status in the 18 deleted families of Duchenne and Becker muscular dystrophy. Methods Deletion analysis of the probands was performed by multiplex polymerase chain reaction (PCR) to amplify 9 dystrophin exons described by Chamberlain. Polymorphism linkage analysis was made on DNA with PCR amplification using primers of intragenic short tandem repeat sequences (STR44, STR45, STR49 and STR50), primers of 5′ end (5′DYS II) and primers of 3′end (MZ18, MZ19) in the members of the families. Gene dosage analysis was performed and DQ value was calculated. Results Both of deletions of exons and STR allelic fragments adjacent to the deleted exons were determined in the probands. STR allelic fragments of 6 pairs of heterozygotes, 2 pairs of homozygotes and 11 hemizygotes were detected at those loci in all of the female relatives. 13 female relatives in deleted families were assayed with gene dosage analysis. In 9 /13 female relatives DQ value was in the range of single copy and carrier status was ascertained.Conclusion Repeat sequence polymorphism as well as gene dosage analysis can potentially be used in carrier detection in the deleted families of Duchenne and Becker muscular dystrophy.

Objective To explore the clinical significance of prenatal ultrasonic diagnosis of fetal nephrohydrosis and its prognosis.Methods Prenatal ultrasanography was performed on 9526 women at more than 20 weeks gestation,and 162 women whose anteroposterior diameter of the renal pelvis was≥8 mm were included in this study.The grade of fetal nephrohydrosis was classified according to Grignon grading method.The changes in fetal nephrohydrosis were observed regularly until delivery.Results(1)The incidence of fetal encephalic fluid and Grignon grade:there were 162 fetuses with fetal nephrohydrosis among 9526 fetuses,with an incidence of 1.7%.The numbers of fetuses with Grades 1 to 5 were 71,59,7,3 and 22,respectively.(2)The distribution in gestafional weeks of fetal nephrohydrosis:generally,fetal nephrohydrosis was first diagnosed at(33±5)weeks,and the maximum degree of nephrohydrosis was observed at(36±3)weeks.One hundred and ten fetuses with nephrohydrosis recovered during the process of gestation,at about(37±4)weeks.(3)Poor terminations of pregnancy:there were 40(25%)fetuses with poor terminations among totally 162 cases.Among these 40 fetuses,3(27%)were first diagnosed at 20 to 24 weeks within 11 fetuses,6(24%)were first diagnosed at 25 to 28 weeks within 20 fetuses,14(26%)were first diagnosed at 29 to 32 weeks within 53 fetuses,11(23%)were first diagnosed at 33 to 36 weeks within 48 fetuses,and 6(20%)were first diagnosed at 37 to 40 weeks within 30 fetuses,The results demonstrate that high grade of fetal nephrohydrosis according to Grignon grading method indicates a poorer prognosis of the fetus.(4)Follow-up results:there were 122(75%,122/162)live and healthy neonates in total,including 110 neonates whose nephrohydrosis recovered prior to birth and 12 neonates whose nephrohydrosis recovered within 1 week after birth.These neonates all developed well without any urinary sequela within the first 2 years.There were also 20(12%,20/162)hydronephrotic neonates who were still diagnosed as nephrohydrosis within 1 week after birth,including 11 fetuses graded below Grignon grade 3,whose nephrohydrosis recovered 3 to 12 months after birth and who developed well without any urinary symptoms.Conclusions The study demonstrates that hydronephrotic fetuses who are diagnosed at earlier gestational weeka and with higher grading have poore prognosis.The Grignon grading method can be used in the prenatal evaluation of fetal nephrohydrosis to predict the prognosis of the fetus.

BACKGROUND: Optical mapping technique is a novel electrophysiological detection method in which voltage-sensitivity dye is medium and silicon photoelectrical diode transforming technology is characteristic, used for analyzing the spatial-temporal distribution of membrane potential in complex neural system.OBJECTIVE: To observe the spatial-temporal changes of brainstem auditory electro-stimulation evoked potential by using optical mapping technology, so as to probe into the influence of γ-Aminobutyric acid (GABA) and γ-GABA receptor antagonist-bicuculline (BMI) on auditory evoked potential.DESIGN: Randomized controlled study.SETTING: Aging Medicine Research Institute of Military General Hospital and E.T.N Department of Japanese Kansai Medical University.MATERIALS:This study was conducted at E.T.N Department of Japanese Kansai Medical University from May to November 2002. Totally 100 ddy/ddy rats, with age of 0-5 days, clear grade, either gender were selected.METHODS: All rats were put to death after cryo-anaesthetized, and brainstem was cut into slices under frozen state so as to remain activity.One side of brainstem slice was connected with the residual end of untraumatic auditory nerve, and slices were put on organic glass plate with the bottom covered with siliac gel and fixed by tungsten filament of 30 μm wide. The residual end of untraumatic auditory nerve was stimulated by tungsten electrode, meanwhile the evoke potentials were recorded at cochlea nuclei and vestibule nuclei. In control group slices were incubated in artificial CSF for 20 minutes, which added with 50 μmol/L γ-GABA in experimental group for observing the influence of γ-GABA on brainstem auditory evoked signals; or alternatively incubated with 50 μmol/L g-GABA and 200 μmol/L for 20 minutes for observing the influence of BMI on brainstem auditory evoked signals. Stimulation was positive rectangle-shape impulse with electric current of 5 mA and frequency of 0.1 Hz, lasting period of 5 ms, the onset time of electric stimulation was set at 89.9 ms.Brain stem slices were stained with electric-sensitivity dye of NK3041 and 16×16 pixel silicon photoelectrical diode device was used to record the auditory nerve stimulation evoked optical signals.fluence of γ-GABA and BMI on optical signals.RESULTS: Totally 100 ddy/ddy rats enrolled in this study and 56 died Character of brainstem auditory electrical-stimulation evoked optical signals: Spatial-temporal changes of auditory evoked optical signals were recorded. The latency of optical signals was (4.63±1.01) ms cochlea nuclei and (6.00±0.89) ms at vestibule nuclei of the same side. Each optical sigual can be divided into two parts: Quick peak-potential and slow long-term potential. The onset phage of quick potential displayed pre-synaptic property, while later phage displayed post-synaptic property; the slow long-term reaction due to γ-GABA and BMI administration: 50 μmol/L γ-GABA was added into the perfusion fluid so as to reduce the amplitude of auditory evoked brainstem neuronal signals in maximum degree, the onset latency of quick potential did not prolonged, but amplitude slightly descended; while the latency and amplitude of later phage were obviously reduced. But such changes could be partly resumed due to the 200 μmol/L dicentrine administration.CONCLUSION: Auditory electro-stimulation evoked potentials can be detected by using multiple-site optical mapping system, which displaying spatial-temporal property. γ-GABA can obviously attenuate the amplitude of auditory electro-stimulation evoked brainstem signals, but such effect can be partly but not completely attenuated by γ-GABA A receptor antagonist-BMI, which implying that besides γ-GABA A receptor, such inhibition of γ-GABA energetic neurons on auditory electro-stimulation evoked potentials can be achieved by other γ-GABA subtype receptors.

Objective To explore extracellular signal-regulated kinase ( ERK1/2 ) expression in the role of curcumin inhibited staurosporine (STS)-mediated neurons toxic injury through added PD098059, and to clarify ERK1/2 mediated inhibitory role of curcumin on STS-induced neurons toxic injury.Methods The neurons toxic injury model of primary cultured hippocampal neurons was established by STS.The experiment was divided into six groups:normal control group, STS model group, PD098059 +STS model group, curcumin +STS pretreatment group, curcumin+PD098059+STS treatment group and curcumin treatment group.The cell viability were determined by MTT method, lactate dehydrogenase (LDH) release rate, cell toxicity were detected, nuclear shape were observed by DAPI staining, and ERK1/2 expression were detected by Western blot method.Results The cell viability of curcumin +STS pretreatment group was significantly higher than STS model group ( P <0.001 ); the cell viability had no significant difference between PD098059 +STS model group and curcumin +PD098059 +STS treatment group;compared with curcumin +STS model group , the cell viability of curcumin +PD098059 +STS treatment group was significantly decreased ( P<0.001 ).LDH results showed that the nerve cell toxicity of curcumin +STS pretreatment group was obviously less than STS model group (P<0.001).The cell nuclear shape showed typical apoptosis morphological characteristics in STS model group, and curcumin inhibited the effect of STS mediated-neuronal apoptosis.ERK1/2 protein expression in curcumin +STS pretreatment group significantly increased compared with STS model group ( P <0.001 ) .Conclusion Curcumin inhibited STS-mediated neurons toxicity injury by up-regulating ERK1/2 expression.After PD098059 blocking the nerve cells ERK1/2 synthesis, the inhibitory action of curcumin failed to implemented, which illustrated that ERK1/2 mediated curcumin to inhibit STS-induced neuronal toxic injury.

Objective To explore the significance of case managers using WeChat platform in the treatment process and extended care for patients with laryngeal cancer.Methods From November 2014 to May 2015, 30 patients with partial resection or total resection of laryngeal cancer in the department of head and neck surgery were selected as a research subjects.Case managers provided fully supports,such as establishing WeChat group,replying online consultations,popularizing health education knowledge,providing psychological counselings.Psychological assessment of the patients was conducted by social support scale(SSRS)and anxiety scale(SAS)in the beginning of case management.Results comparison was performed before and after three months intervention.Results The score of subjective support of patients before intervention was (25.13 ±4.81)points,which after intervention was (28.33 ± 4.41)points.The score of support availability before intervention was (7.20 ±1.99)points,which after intervention was (35.27 ±5.90)points.The differences were statistically significant(t =-2.685,-5.280,2.917,all P <0.05).Conclusion The WeChat platform mediated by the case management is effective in the management of the psychological health of patients with laryngeal cancer,which can effectively promote the rehabilitation of patients.

Purpose It is still undetermined about the application of regular ultrasound (US) combined with ultrasonic elastography (UE) in diagnosing superficial lymph nodes disease.Therefore our aim is to evaluate the value of US combined with UE in diagnosing superficial lymph nodes disease.Materials and Methods The ultrasonography and pathological data of 214 patients (with 266 superficial lymph nodes) were analyzed retrospectively.The US and UE data were compared with the pathologic results.Results The elastic classification of reactive hyperplastic and lymphomatous lymph nodes were no greater than grade 3,respectively;that of malignant metastatic lymph nodes was greater than grade 4.The accuracy of UE or US+UE was higher in diagnosing reactive hyperplastic lymph nodes and metastatic lymph nodes' nature compared with US (P＜0.05).The sensitivity and accuracy of US+UE in diagnosing malignant superficial lymph nodes were higher than those of US (90.13％ vs.72.37％;86.09％ vs.68.05％,P＜0.05).Compared with UE alone,those of US+UE were also higher (90.13％ vs.77.63％;86.09％ vs.72.56％,P＜0.05).Conclusion The accuracy of differential diagnosis of benign or malignant superficial lymph nodes can be improved by US+UE method.

Objective To explore the dynamic changes of D-dimers during pregnancy and early puerperium (within 3 days postpartum). Methods A retrospective study was performed among 8 367 healthy women who had term singleton delivery in Women′s Hospital, School of Medicine, Zhejiang University from January 2007 to December 2014. D-dimers concentrations during pregnancy and early puerprium of all the cases were collected. Data of 21 065 D-dimers tests were assigned to 5 groups according to the time of sampling, including early pregnancy (≤12 gestation weeks), middle pregnancy (12-28 gestation weeks), late pregnancy (>28 gestation weeks), 1 postpartum (within 48 hours postpartum) and 2 postpartum (48-72 hours postpartum). The D-dimers concentrations in different groups were compared. The effect of delivery mode on D-dimers of early pureperium was analyzed. The correlation between D-dimers and the thromboembolic disease was also explored. In this study, Student′s t-test and Wilcoxon rank sum test were used for statistical analysis. D-dimers concentration≤0.5 mg/L was used as the normal range. Results (1) D-dimers concentrations during pregnancy were higher than the non-pregnant women (P<0.01), but there was no statistical difference between early pregnancy and late pregnancy (P=0.820). D-dimers concentration in the 1 postpartum group was higher than that of early pregnancy group or late pregnancy group (P<0.01). But in the 2 postpartum group, it was lower than early pregnancy, late pregnancy and 1 postpartum groups. (2)D-dimers in cesarean section cases was significantly higher than in vaginal delivery cases in each period of pregnancy and early pueprium.(3)The 95%CI of D-dimers in early pregnancy, late pregnancy, 48 hours after vaginal delivery, 48-72 hours after vaginal delivery, ≤48 hours after cesarean section, 48-72 hours after cesarean section were 0.58-8.28, 0.47-11.52, 1.04-9.59, 0.87-5.22, 1.07-11.58 and 1.00-6.23 mg/L, respectively.(4)In 6 cases with thromboembolic disease, D-dimers was 6.89-19.89 mg/L, with the mean value of 13.66 mg/L. It was significantly higher than normal range. In 3 cases, all after cesarean section, with lower extremity vein thrombosis within 48 hours postpartum, the D-dimers concentrations, 9.77, 8.65 and 6.89 mg/L respectively, were in the 95%CI of the study population after cesarean section. Conclusions D-dimers concentration of 0.5 mg/L is not suitable for venous thromboembolism screening during pregnancy. D-dimers concentration in pregnancy and early puerprium is higher than non-pregnancy. It increases in the very early period postpartum and decreases with time. D-dimers should not be a routine screening test to exclude thromboembolic disease in pregnant women without high risk factors and clinical manifestation of thromboembolic disease.

OBJECTIVETo investigate the effect of epidermal stem cells from human hypertrophic scar (HS-ESCs) on the skin wound healing in nude mice.

METHODS40 mice were randomly divided into two groups as experimental group (n = 20) and control group (n = 20). Wounds, 1 cm in diameters, were made on every mouse back. The wounds were treated with HS-ESCs and erythromycin ointment in experimental group, or only with erythromycin ointment in control group. The wound healing was observed during the following 14 days. The expression of collagen-I, collagen-III, epidermal growth factor (EGF), fibroblast growth factor (FGF2) , transforming growth factor (TGFbeta1, and TGFbeta2) were studied.

RESULTSThe wound healing time in the experimental group was (20.8 +/- 0.84) d, which was (25.6 +/- 0.89) d in the control group. HE staining revealed that the extent of vascularization in the experimental group was 11.60 +/- 0.55, while it was 8.04 +/- 0.33 in the control group. Immunochemistry analysis showed the expression of collagen-I, collagen-III, EGF, FGF2, TGFbeta1, and TGFbeta2 in the experimental group were significantly higher, compared with those in control group (P < 0.05).

CONCLUSIONHS-ESCs may promote wound healing through enhancement of the vascularization of the wound tissue and the expression of growth factors.

Animals ; Cicatrix, Hypertrophic ; pathology ; Epidermis ; cytology ; Female ; Humans ; Male ; Mice ; Mice, Nude ; Skin ; injuries ; Stem Cell Transplantation ; Stem Cells ; Wound Healing

A new method for the determination of organic acids from mainstream cigarette smoke using ultra low temperature solvent extraction-comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry(GC×GC-TOF/MS)was established. The mainstream smoke was directly trapped by ethyl ether with ultra low temperature solvent extraction device, and cleaned up with liquid-liquid extraction. The concentrated extracts were further derived by N,O-bis(trimethylsily)trifluoroacetamide (BSTFA). The good separation of silanized product was achieved by the column set of DB-1 (30 m × 0. 25 mm, 1. 0 μm) as the 1st column and DB-wax (1. 5 m × 0. 1 mm, 0. 1 μm) as the 2nd column with modulation period of 6 s and scan range of m/z 45-450 . The results showed that linearity correlation coefficients were larger than 0 . 99 , and the average recoveries were between 80. 17% and 107. 81% with the relative standard deviations (RSD) in the range of 0 . 4%-12 . 1% ( n=5 ) . The detection limit and the quantitation limit were 1 . 3-24 . 5 μg/kg and 4. 1-77. 1 μg/kg, respectively.

Objective To assess the relationship between the serum IgG antibody titer of pregnant women and the hemolytic dis‐ease of newborn(HDN) .Methods Using microcolumn gel coombs card assay method to determine titer of 255 cases of couples an‐tenatal serum IgG antibody and ABO blood group .Results with 195 cases of ABO‐incompatible couples ,the positive rate of abnor‐mal serum IgG antibody(≥64)was 93 .8% .The titer of anti‐A/B IgG :in 12(6 .2% ) cases was <64;in 12(6 .2% )cases was 64;in 60(30 .8% )cases was 128 ;in 39(20% )cases was 256 ;in 45(23 .0% )cases was 512 ;in 27(13 .8% ) cases was 1 024 .There were no statistical differences between IgG anti‐A(B) titers distribution between A/O blood group matching and B/O blood group matching (χ2 =4 .361 ,P=0 .499) ,IgG anti‐A( B) titers was higher in AB/O blood group matching .Conclusion we can take early and effec‐tive prevention ,treatment ,reducing the incidence of HDN by the determination of prenatal serum IgG antibody titers in ABO‐in‐compatible couples ,which is important for the population of eugenics .