Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

AIMTo establish a comprehensive HPLC analytical method of Huanglianjiedu decoction.

METHODSThis study was performed by HPLC-UV/MS to identify the chemical constituents of the whole and individual herbs of the "Huanglianjiedu decoction". Zorbax Extend C18 (150 mm x 4. 6 mm ID, 5 microm) column was used; the mobile phase was composed of acetonitrile (A) and water (B, with 0.5% acetic acid) with gradient elution; the flow rate was 1.0 mL x min(-1) and the column temperature was setup at 25 degrees C. The detection wavelength was 254 nm.

RESULTSThe chromatogram of Huanglianjiedu decoction showed 21 main peaks. Peaks 1, 2, 5 and 18 were from Gardenia jasminoides Ellis, Peaks 8, 13, 14, 15, 16, 17, 19 and 21 from Scutellaria baicalensis Georgi. While 10 from Coptis chinensis Franch and 20 from Phellodendron amurense Rupr., Peaks 3, 4, 6, 9, 11 and 12 came from them together. Peak 7 presented in the chromatograms of the herbs except Gardenia jasminoides Ellis. By comparison of the retention time, the on-line UV spectra and MS spectra, 11 peaks were identified as 5 (geniposide), 9 (jatrorrhizine), 10 (coptisine), 11 (palmatine), 12 (berberine), 13 (baicalin), 15 (oroxin A), 17 (wogonoside), 19 (baicalein), 20 (obaculactone), 21 (wogonin), then eight of them were quantified by HPLC-UV.

CONCLUSIONThe method could represent the characteristics of Huanglianjiedu decoction, and it could be used to evaluate the quality and quantity of Huanglianjiedu decoction. It distinguished between Coptis chinensis Franch and Phellodendron amurense Rupr. by HPLC for the first time.

Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gardenia ; chemistry ; Mass Spectrometry ; methods ; Phellodendron ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Scutellaria baicalensis ; chemistry ; Spectrophotometry, Ultraviolet ; methods

Objective To investigate the inhibitory effects of chitosan nano-particles loaded tetrandrine (Tet) on proliferation of cultured pterygium fibroblasts.Methods The deoxycholic acid-modified chitosan (DAMC) derivative was synthesized through amidation reaction,and their properties were investigated.The viability of human pterygium fibroblasts (HPF) was evaluated by cell counting kit-8 (CCK-8) assay after cells were interacted with Tet/DAMC nano-particles on day 1,3 and 5.Results The synthesized chitosan derivative and Tet formed drug-loaded nano-particles,and the agent-loading capacity was approximately 76％,and the sizes of agent-loaded nano-particles were 50-500 nm,with Zeta potential values being positive.The result of in vitro drug release experiment indicated that the nano-particles constantly released Tet in a controlled manner within 48 h.The viability of HPF in Tet group was (60.70 ± 2.30) ％,(50.22 ± 2.35) ％,(21.99 ± 2.07) ％ on day 1,3,5,respectively,but the corresponding data in Tet/DAMC group was (79.77 ±2.09)％,(63.24 ±2.83)％,(40.28 ± 1.19)％,respectively.The CCK-8 assay demonstrated that the Tet/DAMC nano-particles could inhibit HPF proliferation,and presented lower toxicity than Tet alone.Conclusion Chitosan nano-particles loaded tetrandrine exhibits a sustained agent-release behavior,which has obvious inhibitory effects on the proliferation of human pterygium fibroblasts,but its cytotoxicity is significantly lower than the original drug's,thereby possessing a great promise for improving the outcome of Tet for the prevention of pterygium recurrence.

OBJECTIVETo explore the antifertility effect and safety of 30% ethanol retro-injection into the vas deferens of the rat.

METHODSThirty Sprague-Dawley male rats, 3 m of age and (200 +/- 20) g in weight, were equally randomized into an experimental group and a control group. The former received 30% ethanol (0.5 ml) and the latter 0.9% sodium chloride (0.5 ml), both retro-injected into the vas deferens. Pregnancy rates were obtained through pregnancy tests with 60 Sprague-Dawley female adult rats 1.5 m and 3 m after the injection. All the male rats were sacrificed three months later, and tests were done for the rates of sperm motility and deformity as well as for the apoptosis of spermatogenic cells with TUNEL.

RESULTSThe 1.5 m pregnancy rate was 0 and the 3 m sperm motility and pregnancy rates were (0.32 +/- 1.12)% and (0.58 +/- 1.27)%, significantly decreased (P < 0.05) as compared with those of the control group, which were (80.62 +/- 2.68)%, (70.68 +/- 1.62)% and (86.62 +/- 1.68)%, respectively. While the 3 m sperm deformity rate in the experimental group was (78.26 +/- 1.08)%, increased significantly (P < 0.05), and the apoptosis index (AI) of spermatogenic cells was (7.63 +/- 1.16)% as compared with (5.62 +/- 1.32)% of the control group, with no significant difference between the two groups (P > 0.05).

CONCLUSIONRetro-injection of 30% ethanol into the vas deferens of the rat produces significant antifertility effect on rats, but has no significant influence on their spermatogenic cells.

Animals ; Apoptosis ; Epididymis ; drug effects ; Ethanol ; administration & dosage ; pharmacology ; Female ; Male ; Pregnancy ; Pregnancy Rate ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects ; Spermatids ; drug effects ; Testis ; cytology ; Vas Deferens ; drug effects

Adrenal Cortex Hormones ; therapeutic use ; Adult ; Embolism, Fat ; drug therapy ; etiology ; Evidence-Based Medicine ; Fractures, Bone ; complications ; Humans ; Male ; Treatment Outcome

OBJECTIVETo identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level.

METHODSIn two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family.

RESULTSA nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family.

CONCLUSIONThe mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.

Alternative Splicing ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; China ; Chromosomes, Human, X ; genetics ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Linkage ; Humans ; Male ; Membrane Transport Proteins ; Mutation ; Osteochondrodysplasias ; genetics ; pathology ; Pedigree ; Transcription Factors

OBJECTIVETo explore the relationship between physical and biological effects of alternating magnetic field and study the influence of the magnetic field on the reproductive function of murine testes.

METHODSThirty ICR mice were randomized into 5 groups: normal control, X-ray radiation, weak magnetic field (1000 Hz), 1 h strong magnetic field and 2 h strong magnetic field (2000 Hz). The mice were sacrificed at 7 days after the exposure for the analysis of testicular sperm motility, observation of histopathological changes in the testis by HE staining and evaluation of the changes by modified Johnsen grade criteria.

RESULTSThe rates of sperm motility were (42.37 +/- 10.24)% in the normal control group, (39.00 +/- 12.35)% in the X-ray radiation group, (36.00 +/- 17.28)% in the weak magnetic field group, (10.72 +/- 5.67)% in the 1 h strong magnetic field group and (4.44 +/- 2.87)% in the 2 h strong magnetic field group, respectively. Johnsen's scores decreased and the testis damage increased in a dose- and time-dependent manner.

CONCLUSIONMagnetic field, either strong or weak, may damage the testis function by inducing injury to seminiferous tubules and Leydig cells, thickening of the basal membrane, derangement, exfoliation, massive apoptosis and necrosis of spermatogenic cells in the lumen, situation of the epididymis, and consequently the absence of sperm.

Animals ; Electromagnetic Fields ; adverse effects ; Leydig Cells ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Sperm Motility ; Testis ; cytology ; pathology ; radiation effects

OBJECTIVEX linked spondyloepiphyseal dysplasia tarda (SEDL) is heritable osteochondrondysplasia characterized in affected males by disproportional short stature with short neck and trunk resulting from a growth defect of the vertebral bodies, accompanied by barrel chest and degenerative osteoarthropathy of hip joints. This progressive skeletal dysplasia is caused by the SEDL gene located approximately 100 kb centromeric of DXS16 at Xp22. The disorder usually manifests in late childhood without systemic complications, and generally female carriers of SEDL are asymptomatic. So the diagnosis of potential carriers and presymptomatic patients is almost impossible. This study aimed to establish methods of gene diagnosis for finding out potential carriers and presymptomatic patients.

METHODSThe blood samples were collected from 21 individuals in a large Chinese pedigree with SEDL. Microsatellite marker DXS16 was selected for linkage analysis. In order to confirm the allele of DXS16 linked to the pathogenic SEDL gene, polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) were used to examine the variability of the lengths of DXS16, and linkage analysis was performed for the diagnosis of potential carriers and presymptomatic patients. Then the pathogenic mutation of the SEDL gene in the family was identified by bi-directionally direct sequencing of PCR products amplified for each of the four coding exons as well as their exon/intron boundaries. The potential carriers and presymptomatic patients were also diagnosed in this way.

RESULTSSix young individuals (IV(14), IV(19), IV(21), IV(23), V(4), V(7))who wanted to know whether they were carriers or presymptomatic patients were diagnosed by linkage analysis. Four females of them (IV(14), IV(19), IV(21), V(7)) were determined being carriers because they carry the allele of DXS16 which links the pathogenic SEDL gene, and the other two (IV(23), V(4)) being normal individuals for their alleles of DXS16 linked with wild SEDL gene. DNA sequencing identified that the pathogenic mutation of SEDL gene in the family, which was a nucleotide substitution of the splice-acceptor site in intron 2, IVS2 -2 A-->C. This is a novel mutation in the SEDL gene. Four female individuals (IV(14), IV(19), IV(21), V(7)) carried the mutation; individuals IV(23) and V(4) carried the wild SEDL gene. The results of diagnosis of linkage analysis coincide completely with that of DNA sequencing.

CONCLUSIONLinkage analysis is a simple, rapid and inexpensive gene diagnosis method for SEDL and its accuracy was the same as DNA sequencing. Each of linkage analysis and DNA sequencing can be used to diagnose SEDL, which is very helpful for finding potential carriers and presymptomatic patients.

Base Sequence ; Carrier Proteins ; genetics ; Chromosome Mapping ; Female ; Genetic Diseases, X-Linked ; genetics ; Humans ; Male ; Membrane Transport Proteins ; Molecular Sequence Data ; Mutation ; Osteochondrodysplasias ; genetics ; Pedigree ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Transcription Factors

AIM: To analyze the clinical effects of preoperative intravitreal Conbercept combined vitrectomy for proliferative diabetic retinopathy ( PDR) patients.?METHODS:From June 2014 to May 2017, 57 patients (65 eyes) diagnosed with PDR. The patients were divided into two groups according to whether received preoperative intravitreal conbercept: intravitreal group ( 27 cases, 31 eyes), control group (30 cases, 34 eyes). Intravitreal group was treated with 0. 05mg ( 0. 05mL ) conbercept intravitreal injection 3d before vitrectomy, while control group was treated with vitrectomy alone. The overall surgical time, intraoperative bleeding, use of endodiathermy, iatrogenic retinal hole, and silicone oil, postoperative vitreous hemorrhage and the best corrected visual acuity were recorded and analyzed.?RESULTS: The average surgical time of intravitreal group was lower ( P < 0. 05 ), while intraoperative hemorrhage rate, rate of endodiathermy application, iatrogenic hiatal incidence, rate of silicone oil application, incidences of recurrent vitreous hemorrhage of intravitreal group were lower than those of control group ( all P<0. 05 ) . Intravitreal group got better postoperative best corrected visual acuity than control group (P<0. 05).?CONCLUSION: Intravitreal conbercept for proliferative diabetic retinopathy before 25G vitrectomy decreased surgical complications, reduced the surgical time and postoperative vitreous hemorrhage, and improved the potoperative best corrected visual acuity.

Objective To compare the safety and efficacy of 23-Gauge pars plana vitrectomy (PPV) and PPV combined with scleral buckling-PPV (SB-PPV) in proliferative rhegmatogenous retinal detachments with inferior breaks.Methods Retrospectively nonrandomized clinical case study was conducted in 70 patients with proliferative rhegmatogenous retinal detachment associated with inferior breaks between January 2013 and December 2016,including 39 eyes receiving SB-PPV procedures as the SB-PPV group and 31 eyes undergoing PPV procedures as the PPV group.And anatomical success rate for one procedure,lens trauma rate,retinotomy rate,postoperative best corrected visual acuity (BCVA) outcome and intraocular pressure (IOP) were recorded and analyzed.Results The anatomical success rate for one procedure was 92.3％ (36/ 39) in the SB-PPV group and 74.2％ (23/31) in the PPV group,and the difference was statistically significant (P ＜ 0.035).The lens trauma rate was 2.6％ (1/39) in the SB-PPV group and 19.4％ (6/31) in PPV group,with the difference being statistically significant (P ＜0.05).The retinotomy rate was 5.1％ (2/39) in the SB-PPV group and 32.3％ (10/31) in the PPV group,and the difference approached statistically significance (P ＜0.05).There was no significant difference in the postoperative BCVA and IOP between the two groups (both P ＞ 0.05).Conclusion SB-PPV can increase the anatomical success rate for one procedure in patients with rhegmatogenous retinal detachment associated with inferior breaks,and reduce retinotomy rate and lens trauma rate.