Objective To explore the long term effects of gestational diabetes mellitus (GDM)on offsprings of affected women.Methods One hundred and twenty-four singleton pregnant women with GDM,who delivered in Department of Obstetrics and Gynecology,Peking University First Hospital from June 14,2006 to December 31,2007,were enrolled as the study group.Ninety-eight singleton pregnant women with normal glucose metabolism who delivered at the corresponding period were enrolled as the control group.The follow-up study was performed from November 6,2010 to January 31,2011 on their offsprings.Anthropometry indexes,including height,weight,waist circumference,systolic and diastolic blood pressure (DBP),triceps skin fold (TSF) and sub-scapular skin fold (SSF) were measured.According to the development standard of children less than five years old issued by World Health Organization in 2006,there were six detailed evaluation indicators including length/height for age z-score (HAZ),weight for age z-score (WAZ),weight for length/ height z score (WHZ),body mass index (BMI) for age z-score (BAZ),triceps skin folds for agez-score (TSFZ) and sub-scapular skin folds for age z-score (SSFZ).Chi-square,t-test or variance analysis were applied.Results (1) No statistical difference on age,birth weight,sex,height,weight,BMI,waist circumference,blood pressure,TSF and SSF was found between offsprings of study and control group (P＞0.05).(2) Offsprings in both groups were further divided into macrosomia and non-microsomia subgroups,i.e.GDM macrosomic subgroup (n =15),GDM nonmacrosomic subgroup (n=109),control macrosomia subgroup (n=6) and control non-macrosomia subgroup (n=92).Significant difference was shown amont the four subgroups in weight [(19.8±3.9),(17.0±1.9),(17.7±1.7)and (17.2±1.7) kg,F=6.238,P＜0.001],BMI (17.6±2.6,16.0±1.2,16.6±1.1 and 16.2±1.1,F=5.901,P＜0.001),waist circumference [(53.6±5.3),(49.9±2.7),(50.9±3.3) and (50.4±0.9) cm,F=5.307,P＜0.001],WHZ (1.40±1.44,0.45±0.81,0.88±0.75 and 0,60±0.87,F=5.269,P=0.002),HAZ (1.22±0.78,0.47±0.82,0.98±0.74 and 0.50±1.00,F=3.668,P=0.013),WAZ (1.68±1.23,0.58±0.79,1.15±0.85and 0.71±0.93,F=7.361,P＜0.001) and BAZ (1.41±1.52,0.42±0.84,0.81±0.76 and 0.60±0.90,F=5.210,P =0.002).While comparing between each two subgroups,there was statistical difference between GDM macrosomia subgroup and GDM non macrosomia subgroup,and between GDM marosomia subgroup and control non-macrosomia group.(3) While comparing GDM macrosomia and GDM non-macrosomia subgroup with control group [weight (17.2±2.5) kg,BMI (16.2± 1.4),waist circumference (50.5 ± 3.6) cm,DBP (55.2 ± 6.9)mm Hg,SSF(6.1 ± 1.8) mm,WHZ (0.62±0.87),HAZ (0.53±0.99),WAZ (0.73±0.92),BAZ(0.61±0.89)],the weight (F=9.283,P＜0.001),BMI (F=8.707,P＜0.001),waist circumference (F=7.934,P＜0.001),DBP(F=3.123,P=0.046),SSF (F=3.499,P=0.032),WHZ (F=7.639,P＜0.001),HAZ(F=4.709,P=0.010),WAZ (F=10.302,P＜0.001) and BAZ (F=7.689,P＜0.001) was higher in GDM group than the control group.(4) The proportions of overweight and obesity were higher in GDM macrosomia subgroup than in GDM non-macrosomia and control groups [overweight:9/15 vs24.8％ (27/109) and 24.5％ (24/98),x2 =8.870,P=0.012; obesity:5/15 vs 7.3％(8/109) and 9.2％(9/98),x2=10.083,P=0.006].If all subjects were divided into macrosomia and non-macrosomia group,then the proportion of overweight and obesity was higher in the former group [overweight:52.4％ (11/21) vs24.4％(49/201),x2=7.560,P=0.006; obesity:28.6％ (6/21)vs 7.9％(16/201),x2 =9.047,P=0.003].Conclusions GDM may have long term adverse effect on the development of offsprings at three-four-year-old with higher incidence of obesity or high diastolic blood pressure in macrosomic babies of GDM mothers than in non macrosomic babies of GDM mothers or babies of non GDM mothers.

ObjectiveTo investigate the impacts of neural stem cells (NSCs) transplantation on spinal pathology and ultrastructure after spinal cord injury (SCI) in rats and probe into the protective role of tacrolimus (FK506) on neural regeneration.MethodsCompressive SCI at T8 was induced in the adult SD rats,which were randomly assigned to the control group,FK506 group,NSCs group and NSCs + FK506 group.The differences of neural regeneration in each group were compared at days 7,14,28 and 56 after injury by motor evoked potentials ( MEP),HE staining,immunohistochemical staining,ultrastructure observation and image analysis of the myelinated fiber. ResultsThe MEP latency in the NSCs + FK506 group was significantly shorter than that in other groups at day 28 ( P ＜ 0.05 ).HE staining revealed that only local necrosis presented in the NSCs + FK506 group at day 56.More BrdU and NF-200 positive cells were detected with immunohistochemical staining in the other three groups as compared with the control group.Moreover,the positive cells in the NSCs + FK506 group also outnumbered the FK506 group and NSCs group.Electron microscope scan showed edema under the membrane of large myelin sheath in the control group,and classic new myelin sheath and neuraxis in the NSCs + FK506 group at day 56.The regeneration of the nerve fiber in the NSCs + FK506 group was better than that of other three groups (P ＜0.01 ).ConclusionAfter NSCs transplantation for SCI rats,the early combination use of FK506 can improve the pathology and ultrastructure of the regenerative nerve fiber and is conducive to nerve regeneration.

Objective To evaluate clinical efficacy and safety of levornidazole in the treatment of pelvic anaerobic infections. Methods A multicenter randomized controlled clinical study was conducted to evaluate clinical efficacy and safety of levornidazole. One hundred and fourty-three patients with pelvic anaerobic bacteria infection were classified into 70 cases treated by levornidazole in study group and 73 cases treated by Ornidazole in control group. Those patients in two groups were both administered at a dose of 0. 5 g twice daily for 5 - 7 days. The rate of clinical efficacy, bacteria clearance and adverse effect were recorded and compared between two groups. Results At the endpoint, the rate of clinical efficacy were 80% (56/70) in study group and 81% (59/73) in control group, which did not reach significant difference (P＞0. 05). The rate of bacteria clearance were 97% (36/37) in study group and 92% (22/24) in control group, which also did not reach significant difference(P ＞0. 05). The rate of adverse reaction of 3% (20/70) in study group was significantly lower than 22% ( 16/73 ) in control group ( P ＜ 0. 05 ). Conclusion It is effective and safe to treat pelvic anaerobic infections with levornidazole and sodium chloride injection.

Objective To explore the impact of pre-implant hearing aid fitting on the early open-set word recognition in children who received a cochlear implant(CI) when they were infants .Methods A total of 289 chil‐dren who received a cochlear implant between 0 .9 and 3 .0 years of age were included in this study .According to pre-implant hearing aid fitting ,participants were divided into hearing aid group and non-hearing aid group .The open-set word identification abilities of the cochlear implant children were evaluated via Mandarin Lexical Neighborhood test(M -LNT) .Results Comparisons of the scores of M -LNT between two groups displayed no significant differ‐ence at the same identification age or duration of CI use .Conclusion The abilities of the early open-set word rec‐ognition between hearing aid group and non-hearing aid group didn’t make significant differences for the CI chil‐dren w ho received a cochlear implant betw een 0 .9 and 3 .0 years old .

OBJECTIVE To analyze mutation of POMT1 gene in a Chinese family affected with congenital muscular dystrophy (CMD). METHODS Peripheral blood samples of the family including one affected and two unaffected individuals, in addition with chorionic villous sample from the fetus, were collected. PCR was used to amplify exons 19 and 20 of the POMT1 gene, and the products were sequenced directly. Based on the result of genetic testing, prenatal diagnosis of the fetus was attained. RESULTS The proband was found to carry a heterozygous missense mutation c.1939G>A (p.Ala647Thr) in exon 19 of the POMT1 gene inherited from the mother and a heterozygous frameshift mutation c.2141delG (p.Trp714Ter) in exon 20 inherited from the father. Prenatal diagnosis revealed that the fetus has carried the c.1939G>A (p.Ala647Thr) missense mutation. With the disease causing mutation, the fetus was predicted to have similar phenotype as its mother. CONCLUSION The compound heterozygous mutations of c.1939G>A (p.Ala647Thr) and c.2141delG (p.Trp714Ter) probably underlie the CMD in this family. Based on the result, prenatal diagnosis may be provided.

OBJECTIVETo explore the feasibility of using PCR-based capillary electrophoresis method to analysis mutation of the TOR1A gene in a family affected with primary torsion dystonia (PTD).

METHODSPeripheral blood sample was collected from proband and amnionic fluid from her fetus for the extraction of DNA. The 5th exon of the TOR1A gene and its flanking sequences were amplified with PCR and analyzed with agarose electrophoresis, fluorescence labeled fragment analysis and Sanger sequencing.

RESULTSFluorescence labeled fragment analysis was performed through capillary electrophoresis, which showed that the proband carried a c.907_909delGAG (p.Glu303del) deletional mutation of the TOR1A gene. The result was verified by Sanger sequencing. The fetus DNA was also found with the same mutation by capillary electrophoresis, inferring that the fetus was probably affected with the disease.

CONCLUSIONThe mutation of c.907_909delGAG of the TOR1A gene was speculated as pathologic cause of proband in this family. Fragment analysis by capillary electrophoresis combined with DNA sequencing is an efficient test for small deletional mutations and feasible for its prenatal diagnosis.

Adult ; Dystonia ; diagnosis ; genetics ; Electrophoresis, Capillary ; Female ; Humans ; Molecular Chaperones ; genetics ; Mutation ; Prenatal Diagnosis ; Sequence Analysis, DNA

OBJECTIVE: To explore the characteristics of PAH gene variants among 113 phenylketonuria patients from Henan Province. METHODS: The 13 exons of the PAH gene were subjected to PCR amplification and direct sequencing. Large fragment deletion and duplication of the PAH gene were detected with a multiple ligation-dependent probe amplification (MLPA) assay. RESULTS: In total 195 point variants and 3 large fragment deletions were detected among the 226 alleles, with the detection rates being 86.28% and 1.33%, respectively. Variants of p.Arg243Gln (18.14%), p.Arg111X (6.19%), p.Arg53His (5.31%), EX6-96A>G (5.31%), p.Tyr356X (4.87%) and p.Val399Val (4.42%) were relatively common. Most of the variants were located in exons 7, 11, 3 and 6. Missense variations were most common. Four novel variations were detected, which included c.1016C>A (p.Ser339Tyr), c.1000T>C (p.Cys334Arg), c.1110G>T (p.Glu370Asp), and IVS6+1G>T. CONCLUSION The PAH gene variations in Henan Province have featured extensive allelic heterogeneity and variety.
China ; Exons ; Humans ; Mutation, Missense ; Phenylalanine Hydroxylase ; genetics ; Phenylketonurias ; genetics ; Point Mutation ; Sequence Deletion

[ ABSTRACT] AIM:To investigate the effects of Chaihu Shugan powder ( CSP) on lipid metabolism and the pro-teins involved in adenosine 5’-monophosphate-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) pathway in the liver tissues of the rats with non-alcoholic fatty liver disease (NAFLD).METHODS: Sprague-Dawley rats were randomly di-vided into normal control ( NC) group, with HFD ( HFD) group and CSP group.The NAFLD models were established by feeding with HFD for 16 weeks in the rats.The rats in CSP group were intragastrically administered with CSP extracts (9.6 g· kg-1 · d-1 ) , and blood and liver samples were collected 16 weeks later.Serum and liver levels of total cholesterol ( TC) and triglyceride ( TG) , and serum levels of alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) were measured using an automatic biochemical analyzer.The histological changes of liver tissues were observed with HE staining, while the lipid deposition was observed with Oil Red O staining.The ultrastructural changes of the liver tissues were observed under transmission electron microscope.Moreover, the protein levels of AMPK, phosphorylated AMPK (pAMPK), SIRT1 and uncoupling protein 2 (UCP2) in the liver were detected by Western blot.RESULTS:The results of HE staining, Oil Red O staining and electron microscopy demonstrated that NAFLD rat model was successfully estab-lished.Compared with NC group, the serum and liver levels of TC and TG, and serum level of AST in model group were markedly elevated ( P<0.01) .Moreover, the protein levels of pAMPK and SIRT1 in HFD group were markedly reduced (P<0.01), whereas UCP2 level was elevated (P<0.01).Furthermore, liver levels of TC and TG, and serum level of AST in GSP group were markedly reduced as compared with HFD group ( P<0.05 ) .The protein levels of pAMPK and SIRT1 were elevated ( P<0.05 ) , whereas the UCP2 level was reduced as compared with HFD group ( P<0.01 ) .The protein level of AMPK between the 3 groups had no significant difference.CONCLUSION: CSP attenuates hepatic lipid disorder and hepatic lipid deposition in NAFLD rats induced by feeding with HFD for 16 weeks, which is associated with the activation of AMPK/SIRT1 pathway.

Community-acquired respiratory infection is the commonest cause of sepsis presenting to emergency departments. Yet current experimental animal models simulate peritoneal sepsis with intraperitoneal (I.P.) injection of lipopolysaccharide (LPS) as the predominant route. We aimed to compare the progression of organ injury between I.P. LPS and intranasal (I.N.) LPS in order to establish a better endotoxemia murine model of respiratory sepsis. Eight weeks old male BALB/c mice received LPS-Escherichia coli doses at 0.15, 1, 10, 20, 40 and 100 mg per kg body weight (e.g. LPS-10 is a dose of 10 mg/kg body weight). Disease severity was monitored by a modified Mouse Clinical Assessment Score for Sepsis (M-CASS; range 0–21). A M-CASS score ≥ 10 or a weight reduction of ≥ 20%, was used as a criterion for euthanasia. The primary outcome was the survival rate (either no death or no need for euthanasia). The progression of disease was specified as M-CASS, body weight, blood glucose, histopathological changes to lung, liver, spleen, kidney, brain and heart tissues. Survival rate in I.P. LPS-20 mice was 0% (2/3 died; 1/3 euthanized with M-CASS > 10) at 24 h. Survival rate in all doses of I.N. LPS was 100% (20/20; 3–4 per group) at 96 h. 24 h mean M-CASS post-I.P. LPS-10 was 6.4/21 significantly higher than I.N. LPS-10 of 1.7/21 (Unpaired t test, P < 0.05). Organ injury was present at 96 h in the I.P. LPS-10 group: lung (3/3; 100%), spleen (3/3; 100%) and liver (1/3; 33%). At 24 h in the I.P. LPS-20 group, kidney injury was observed in the euthanized mouse. At 96 h in the post-I.N. LPS-20 group, only lung injury was observed in 2/3 (67%) mice (Kruskal-Wallis test with Dunn’s, P < 0.01). At 24 h in the post-I.N. LPS-100 group all (4/4) mice had evidence of lung injury. Variable doses of I.N. LPS in mice produced lung injury but did not produce sepsis. Higher doses of I.P. LPS induced multi-organ injury but not respiratory sepsis. Lethal models of respiratory virus, e.g., influenza A, might provide alternative avenues that can be explored in future research.

Objective To investigate the value of next-generation sequencing (NGS) technique for genetic analysis of spontaneous abortion. Methods From January to June 2017, 154 patients who visited the First Affiliated Hospital of Zhengzhou University for spontaneous abortion were enrolled. All abortion tissue samples were analyzed by both NGS combined with short tandem repeat (STR) and single nucleotide polymorphism array (SNP-array). Results of the two methods were compared by Chi-square or Fisher's exact test. Results (1) Chromosomal abnormalities were detected in 109 of the 154 cases (70.7%), including 52 (47.7%) of numerical chromosomal abnormalities, 49 (45.0%) of structural chromosomal abnormalities, six (5.5%) of mosaicism, and two (1.8%) of uniparental disomy (UPD). In those 52 cases of numerical chromosome abnormalities, there were 45 of chromosome aneuploidy and seven of polyploidy. The top three numerical chromosomal abnormalities were 45,X (27.0%, 14/52), trisomy 22 (9.6%, 5/52) and trisomy 16 (7.7%, 4/52). Forty-nine structural abnormality cases carried 67 copy number variations (CNV), including 13 pathogenic CNV (pCNV, 19.4%), 24 variants of unknown clinical significance (35.8%) and 30 benign CNV (44.8%). In those 13 pCNVs, two were responsible for microdeletion and microduplication syndromes. (2) SNP-array was successful in 152 cases, but failed in two (1.3%) due to genomic DNA <200 ng. However, NGS technology was successful in all 154 cases and identified chromosomal abnormalities in the two cases that SNP-array had failed. No statistically significant difference was shown in the detection rate of chromosomal abnormalities between SNP-array and NGS technology [70.4% (107/152) vs 67.5% (104/154), χ2=0.293, P=0.588]. (3) No significant difference in the detection of chromosome aneuploidy (six cases in each group, 3.9% vs 3.9%) and mosaicism (45 cases in each group, 29.2% vs 29.6%) was found between NGS technology and SNP-array. Three cases of polyploidy (69, XXX) and two of UPD were identified by SNP-array, but not by NGS. When combined with STR, NGS was able to detect all three cases of polyploidy (69, XXX). (4) Forty-seven structural abnormality cases detected by SNP-array carried 53 CNVs, and 49 detected by NGS carried 67 CNVs. (5) NGS detected ten, three and one more CNVs than SNP-array did when the genome lengths were 100-<500, 500-<1 000 and ≥1 000 kb, respectively. Conclusions NGS can be used to detect chromosomal aneuploidy and mosaicism that can be identified by SNP-array with fewer limitations on total amount of genome. Moreover, CNVs that fail to be identified by SNP-array can also be detected by NGS. When combined with STR, NGS can effectively detect chromosomal polyploidy. Therefore, NGS could be a potential genetic analysis method for spontaneous abortion and of importance for genetic counseling.