OBJECTIVE: A case of half sibling was determined with multiple genetic markers, which could be potentially applied for determination of half sibling relationship from same father. METHODS: Half sibling relationship was detected by 39 autosomal STR genetic markers, 23 Y-chromosomal STR genetic markers and 12 X -chromosomal STR genetic markers among ZHAO -1, ZHAO -2, ZHAO -3, ZHAO -4, and ZHAO-5. RESULTS: According to autosomal STR, Y-STR and X-STR genotyping results, it was determined that ZHAO-4 (alleged half sibling) was unrelated with ZHAO-1 and ZHAO-2; however, ZHAO-3 (alleged half sibling), ZHAO-5 (alleged half sibling) shared same genetic profile with ZHAO-1, and ZHAO-2 from same father. CONCLUSION It is reliable to use multiple genetic markers and family gene reconstruction to determine half sibling relationship from same father, but it is difficult to determination by calculating half sibling index with ITO and discriminant functions.
Alleles ; Chromosomes, Human, Y/genetics* ; Discriminant Analysis ; Gene Frequency ; Genetic Loci/genetics* ; Genetic Markers ; Genotype ; Humans ; Paternity ; Siblings

ObjectiveTo investigate the quality of life (QOL) of the epilepsy patients and the factors affected the QOL.Methods200 cases were investigated using the Quality of Life in Epilepsy Inventory 10 (QOLEI-10) and the Washington Psychosocial Seizure Investigate (WPSI). 200 healthy persons were chosen as normal control group. ResultsThe QOL of the patient group were significantly poor as compared with that of normal control group(P<0.01). The factors that the patients always faced with were disability in the attack control, short in money, unemployment, restriction of movement, disability in intercommunication, psychological disorder (depress, strain, anxiety, dread, shame feel, cognitional dysfunction, etc.), as well as the difficult to get professional curtains, taking medicine improperly and side effects of the medicine. Conclusions The factors mentioned, which were usually neglected by many doctors, do affect the QOL of epilepsy patients, and hinder the epilepsy treatment effectively.

OBJECTIVE: To evaluate the effect of Kuntai capsule on the gonadotrophin releasing hormone agonist(GnRH-a)-induced perimenopaus symptoms and the sex hormone levels.METHODS: A total of 99 patients with uterine fibroids,adenomyosis or moderate to severe endometriosis who needed the treatment of GnRH-a at Sun Yat-sen Memorial Hospital of Sun Yat-sen University from June 2015 to March 2016 were collected and randomly divided into research group(Kuntai capsule)and control group(Tibolone). GnRH-a was injected once every 28 days and first injection of GnRH-a was administered on the 2 nd to 4 th day of menstrual period or retraction bleeding after surgery.Kuntai capsule or Tibolone was orally taken beginning from the first GnRH-a injection,and the co-administration of Caltrate D-600 and alfacalcidol was given in both groups.The Kupperman scores,sex hormone levels including folliclestimulating hormone(FSH)and estrogen(E_2),and adverse events were recorded.RESULTS: Kuntai capsule kept the perimenopause symptoms at mild level with the slow increase of Kupperman scores,whose effect was significantly superior to Tibolone(P<0.05)after 8 weeks of treatment,especially in paresthesia,nervousness,and formication.The FSH and E_2 levels in both Kuntai and Tibolone groups were obviously decreased when compared with the pre-treatment(P<0.05),and these hormone levels in Kuntai group were comparable to those in Tibolone group.No adverse events occurred in either group. CONCLUSION: In the short-term treatment of GnRH-a,Kuntai capsule exhibits significant alleviating effects on perimenopause symptoms caused by GnRH-a with high safety and few adverse reactions.

BACKGROUNDThe form deprivation (FD) reduces spatial contrasts and induces myopia. Nitric oxide and cyclic guanosine monophosphate (cGMP) are involved in visual signal transmission. This study investigated changes in nitric oxide synthase (NOS) activity and cGMP concentration in ocular tissues in acute and chronic form deprivation myopia.

METHODSGuinea pigs had one eye covered by translucent glass for 7, 14 or 21 days. Untreated litter mates were used as controls. NOS activity and cGMP concentrations in the retinal, choroidal and scleral tissues of FD eyes and control eyes were analyzed by radioimmunoassay after various durations of FD. The expression of NOS subtypes was identified by immunohistochemistry.

RESULTSMyopia was successfully induced in FD eyes after 14 days. Compared with control groups, the retinal NOS activity and cGMP concentrations in the FD eyes significantly increased after 14 and 21 days while the retinal NOS activity in the FD eyes was transiently suppressed by 7 days of FD. The NOS activity and cGMP concentrations of choroid and sclera in the FD eyes were higher than in the control groups at 21 days. The three isoenzymes of nitric oxide synthase were detected in the ocular tissues of guinea pigs.

CONCLUSIONSThe NOS activity and cGMP concentrations were upregulated after chronic FD and the retinal NOS activity was transiently suppressed at acute FD. The function of elevated NOS activity may be mediated by cGMP.

Animals ; Cyclic GMP ; analysis ; Guinea Pigs ; Immunohistochemistry ; Myopia ; metabolism ; Nitric Oxide ; physiology ; Nitric Oxide Synthase ; metabolism ; Refractive Errors ; Retina ; metabolism

ObjectiveTo investigate the morphological characteristics of mitral valve annulus in patients with degenerative mitral valve prolapse with severe regurgitation. MethodsA total of 18 patients with mitral valve prolapse complicated with severe regurgitation and 36 patients with normal mitral valve were selected and analyzed using Philips Epic7C echocardiography equipped with transesophageal 3D probe X7-2T and QLAB MVN software. ResultsThe anterolateral and posterio-medium diameter， anterior-posterior diameter， circumference and area of mitral annulus in patients with mitral prolapse complicated with severe regurgitation were larger than those in the control group （P<0.05）. In addition， the annulus parameters in the middle and late systole were higher than those in the early systole （P<0.05）， while the height of the mitral annulus and the non-planar angle showed no difference between the two groups or among different cardiac cycles （P>0.05）. ConclusionsThe three-dimensional morphology and functional of mitral valve annulus changed dynamically with different phases of cardiac cycle. Three-dimensional esophageal echocardiography could provide accurate and detailed information for degenerative mitral valve prolapse with regurgitation.

AIMTo detect the expression of HSP25 in rat dental follicles both in vivo and vitro, and explore the underlying mechanism of HSP25 on the proliferation and differentiation of rat dental follicle cells (DFCs).

METHODOLOGYImmunohistochemistry was performed to detect the expression of HSP25 in mandibles of postnatal rats on days 1, 3, 5, 7, 9 and 11 in vivo. In vitro, the expression of HSP25 in DFCs was detected by an indirect immunofluorescence assay. Thiazolyl blue tetrazolium bromide (MTT) assay, flow cytometry and alkaline phosphatase (ALP) assay were used to identify the time-course effect mediated by different concentrations of recombinant murine HSP25 of 0, 1, 10, 50 and 100 ng/mL on rat DFCs.

RESULTSExpression of HSP25 was not detected in dental follicles of the rats until day 5 after birth, but became up-regulated in a time-dependent manner till day 11. HSP25 was detected in the cytoplasm of cultured rat DFCs. No significant difference could be observed in the proliferation of DFCs after stimulation with different concentrations of HSP25 on days 1, 2 and 3 (P > 0.05). HSP25 at concentrations of 50 ng/mL and 100 ng/mL up-regulated the ALP activity of DFCs on day 9 (P < 0.05).

CONCLUSIONHSP25-immunoreactivity increased chronologically during the development of dental follicles. The protein had no significant effect on cell proliferation but may play a role in cementoblast/osteoblast differentiation of DFCs.

Alkaline Phosphatase ; analysis ; Ameloblasts ; cytology ; Animals ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Coloring Agents ; Cytoplasm ; ultrastructure ; Dental Sac ; cytology ; growth & development ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; HSP27 Heat-Shock Proteins ; analysis ; physiology ; Odontoblasts ; cytology ; Rats ; Rats, Sprague-Dawley ; Tetrazolium Salts ; Thiazoles ; Tooth Germ ; cytology ; Up-Regulation ; physiology

Fruit ripening is associated with a number of physiological and biochemical changes. They include degradation of chlorophyll, synthesis of flavor compounds, carotenoid biosynthesis, conversion of starch to sugars, cell wall solublisation and fruit softening. These changes are brought about by the expression of specific genes. People are interested in the molecular mechanism involved in the regulation of gene transcription during fruit ripening. Many fruit-specific promoters such as PG, E4, E8, and 2A11 have been characterized and shown to direct ripening-specific expression of reporter genes. AGPase plays the key role in catalyzing the biosynthesis of starch in plants. It is a heterotetrameric enzyme with two small subunits and two large subunits, which are encoded by different genes. In higher plants, small subunits are highly conserved among plant species and expressed in all tissues. And the large subunits are present at multiple isoforms and expressed in a tissue-specific pattern. In fruits, the expression pattern of the large subunits varies with plant species. That made it important to study the transcriptional regulation of the large subunits of AGPase in different plant species. Northern-blot analysis indicates in watermelon, an isoform of the large subunits Wml1 expressed specifically in fruits, not in leaves. The 5' flanking region of Wml1, which covers 1573bp, has been isolated through the method of uneven PCR. And transient expression assay has shown that the 1573bp (named WSP) can direct fruit-specific expression of GUS gene. Our goal in this study was to scan the promoter region for main regulatory regions involved in fruit-specific expression. A chimaeric gene was constructed containing the WSP promoter, the beta-glucuronidase (GUS) structural sequence as a reporter gene and the nopaline synthase polyadenylation site (NOS-ter). The plasmid pSPA was digested with Hind III + Hinc II and promoter fragment of 1573bp (from 180bp to 1752bp) was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-16. The same insert was then cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by HindIII + Bgl II to produce pISPA-16. Three 5'-end deletions of the promoter were obtained and fused to GUS gene in plant transient expression vector pBI426: the 1201bp fragment (from 551bp to 1752bp) was generated by digestion of pBSPA-16 with BamH I + SnaB I, the 898bp fragment (from 854bp to 1752bp) by BamH I + EcoRV. Both fragments were ligated with pBluescript SK(-) digested by BamH I + Sma I, to produce pBSPA-12 and pBS-PA-9. The inserts were cut out with HindmIII + BamH I and ligated with pBI426 digested by Hind III + Bgl II, to produce pISPA-12 and pISPA-9. The 795bp fragment (from 957bp to 1752bp) was generated by digestion of pSPA with Hinc II + EcoR I, promoter fragment was cut out and cloned into Sma I sites of pBluescript SK(-), to produce pBSPA-8. The same insert were cut out with Hind III + BamH I, and ligated with transient expression vector pBI426 digested by Hind III + Bgl II. The 1573bp fragment and three 5'-end deletions were delivered into watermelon leaf, stem, flower and fruit of different development stages (5, 10, 20 days after pollination) via particle bombardment using a biolistic PDS-1000/He particle gun. Bombardment parameters were as follows: a helium pressure of 1200 psi, vacuum of 91432.23Pa, 7 cm between the stopping screen and the plate. Histochemical assay were done on all the tissues bombarded after incubation for 2 days. The 1573bp fragment had the strongest promoter activity, and can induce GUS expression in fruits of 5 and 20 days after anthesis and flowers, but not in fruits of 10 days after anthesis, leaves and stems. Fragments of 1201bp and 898bp can induce GUS expression only in fruits of 20 days after anthesis, and with lower expression levels than 1573bp. Fragment of 795bp was not able to direct GUS expression in any of the tissues bombarded (data not shown). It can be concluded that of the 1573bp, 1201 bp, 898bp Wml1 5'flanking regions include the necessary information directing fruit-specific expression. Deletion from 180bp to 551bp doesn't affect the fruit-specificity of the promoter, but lowered the expression level. There may be some cis-acting elements located in this region, which can enhance external gene expression in later stages of fruit development. Deletion from 854bp and 958bp led to loss of GUS expression. This region includes the necessary information needed for gene expression as well as the regulatory elements for fruit-specific transcription.
Citrullus ; genetics ; Fruit ; genetics ; Gene Expression Regulation, Plant ; genetics ; Promoter Regions, Genetic ; genetics ; Regulatory Sequences, Nucleic Acid ; genetics ; physiology

OBJECTIVETo establish a model of chronic periodontitis (CP) accompanied with chronic obstructive pulmonary disease (COPD) in SD rats and investigate the relationship between chronic periodontitis and COPD.

METHODSEqual gender SD rats were randomly divided into 4 groups, A: control group, B: CP group, C: COPD group, D: COPD with CP group (n = 10, respectively). Each group was subjected to its predesigned intervention to establish a specific disease model. After 10 weeks, animals were sacrificed. The level of alveolar bone loss, lung function measurement, and the histopathological changes of periodontal and lung tissues were examined. The serum tumor necrosis factor (TNF)-α level was detected by enzyme-linked immunoabsorbent assay kits.

RESULTSBleeding index (BI) levels of group A and C were (0.25 ± 0.04) and (1.30 ± 0.25), respectively. Attachment loss was (0.43 ± 0.02) and (0.51 ± 0.02) mm. BI levels in group B and D were significantly higher than those in group A and C. Forced expiratory volume in 0.2 second to forced vital capital ratio (FEV(0.2)/FVC) values in group B, C and D were significantly lower than that in group A. Pulmonary function were worse in group D than that in group C (P < 0.05). The levels of serum TNF-α, a sensitive indicator of both diseases, were increased in all test groups compared with the control, and increased most in group D.

CONCLUSIONSThe chronic periodontitis and chronic obstructive pulmonary disease model was established in SD rat. The chronic periodontitis may be a risk factor for promoting and inducing COPD.

Alveolar Bone Loss ; Animals ; Chronic Periodontitis ; Disease Models, Animal ; Forced Expiratory Volume ; Lung ; Pulmonary Disease, Chronic Obstructive ; Rats ; Tumor Necrosis Factor-alpha

OBJECTIVETo investigate the expression of CXCR4 in cultured human dental pulp cells (HDPC) in vitro and the corresponding ligand SDF-1alpha level of HDPC supernatants stimulated by lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-alpha), and to explore the role of SDF-1alpha on the proliferation and the migration of HDPC.

METHODSThe expression of CXCR4 in HDPC was detected by immunocytochemistry technique and indirect immunofluorescence technique. The culture supernatants of HDPC were collected after HDPC had been simulated by LPS and TNF-alpha of different concentrations for 48h and then the SDF-1alpha level was assayed by quantitative sandwich ELISA. Meanwhile, the effects of recombinant human SDF-1alpha (rhSDF-1alpha) on the proliferation and the migration of HDPC at different concentrations were observed by MTT and Boyden Chamber Assay.

RESULTSCXCR4 was expressed in cytomembrane of HDPC and SDF-1alpha was secreted into their normal cell supernatants with a concentration of (4513.55 +/- 962.92) ng/L. The secretion of SDF-1alpha was both significantly decreased by stimulation with LPS and TNF-alpha (P < 0.05). In addition, rhSDF-1alpha stimulated the HDPC proliferation at the concentrations of 50, 100, 200 microg/L (P < 0.01) and increased the chemotactic migration of HDPC significantly after 9h's incubation with the concentrations of 50, 100 microg/L (P < 0.05).

CONCLUSIONSSDF-1alpha accelerated the proliferation and the migration of HDPC which expressed CXCR4. SDF-1-CXCR4 axis may play a role in repair of pulp injury.

Cell Movement ; Cell Proliferation ; Cells, Cultured ; Chemokine CXCL12 ; metabolism ; Dental Pulp ; cytology ; metabolism ; Humans ; Receptors, CXCR4 ; metabolism

OBJECTIVETo assess the association between leptin gene promoter methylation and serum leptin concentrations in patients with impaired glucose regulation (IGR) and type 2 diabetes mellitus (T2DM).

METHODSMethylation status of leptin gene promoter was determined with methylation-specific polymerase chain reaction. Serum leptin concentrations were determined using enzyme-linked immunosorbent assay.

RESULTSAmong three groups of individuals with different levels of glucose, the methylation rates of leptin gene in IGR and T2DM groups were 43.6 % and 31.5 %, respectively, which were significantly lower than that of healthy subjects (59.2%; Chi-square=22.499 and 5.109, respectively, P<0.05). A lower methylation rate was also observed in T2DM group compared with IGR group (Chi-square=3.962, P<0.05). Leptin levels in both T2DM and IGR groups were elevated compared with normoglycemic subjects, but only T2DM group was significantly higher (q=6.81, P<0.01). Linear regression analysis indicated that serum leptin concentrations has increased along with declining of DNA methylation rate (r=-0.95, P<0.01).

CONCLUSIONLower levels of leptin gene promoter DNA methylation and serum leptin concentrations are associated with the development of diabetes. Measurement of the methylation status of leptin gene promoter and expression can facilitate early intervention of the disease.

DNA Methylation ; Diabetes Mellitus, Type 2 ; genetics ; metabolism ; Female ; Genetic Predisposition to Disease ; Glucose ; genetics ; metabolism ; Humans ; Leptin ; blood ; genetics ; metabolism ; Male ; Middle Aged ; Promoter Regions, Genetic