A new method for noninvasive prenatal diagnosis of fetal sex was developed by using single-cell PEP-PCR techniques. Micromamipulation techniques were used to obtain single fetal cells from 273 maternal blood samples. The genome of single cells was preamplified by PEP and SRY genes were analyzed by PCR method. The SRY genes of 149 samples were detected by the new method among 153 samples carrying male fetus, while 119 out of 120 samples carrying female fetus were proved negative for SRY genes. The sensitivity and specificity of the new method were 97.39% and 99.17% respectively and the correct rate was 98.17%. The new method has the advantage of high sensitivity and specificity in noninvasive prenatal diagnosis of fetal sex and provides the basis of other researches such as sex-linked inherited diseases.
Chromosomes, Human, Y ; Erythroblasts/chemistry ; Fetus/*cytology ; Genes, sry/genetics ; Genetic Diseases, Inborn/diagnosis ; Maternal-Fetal Exchange/genetics ; Polymerase Chain Reaction/methods ; Pregnancy/*blood ; Prenatal Diagnosis/*methods ; *Sex Determination (Genetics)

OBJECTIVE:To investigate the effect of evaluation method on management of prescription. METHODS:A total of 24 000 prescriptions of 12 months were evaluated with both method 1 devised by our hospital and method 2 specified in ordinance of Management of prescription. The administration department intervened physicians’ prescription behaviors according to the evaluation results. The change of data in method 1 was compared with that in method 2. RESULTS: In method 1,the problems of usage and dosage increased a little while other irrational problems lessened largely. In method 2,the average amount of drug categories reduced a little while the other data increased a little. CONCLUSION: The effect of evaluation and intervention in method 1 are superior to those in method 2.

Objective To explore the related factors of insulin sensitivity in infants and toddlers. Methods From January 2006 to May 2008, 246 infants and 120 toddlers who had regular physical check-ups at the Children Healthcare Center of Third Hospital of Peking University were selected in this retrospective study. The homeostasis model assessment insulin resistance index (HOMA-IR), insulin action index(IAI), fasting glucose-to-insulin ratio (FGIR) and HOMA-β cell function (HOMA-β) were measured with the homeostasis model analysis. According to different glucose metabolic situation of the pregnant mothers and diabetes family history, the infants and toddlers were divided into different groups. The above indicators were compared among groups.Results In infants with maternal diabetes, the levels of HOMA-IR and HOMA-β were 3.24 (2.76-4.12) and 164. 00 (114. 44-192.85), higher than those born to mothers without abnomal glucose metabolism [1.51 (0. 86-2.50) and 67.07 (41.83-106.22)], while the levels of IAI and FGIR were lower[-7.18(-7.41-7.02) and 7.31(5.82-8.55) vs -6.41(-6.92-5.85) and 14.84(9.49-24. 79)] (Z=3. 76,3. 35,3. 76 and 3.71, P＜0. 017). Compared with infants of maternal impaired glucose tolerance,the level of HOMA-IR was higher (Z= 3.19, P＜0.017) in infants with maternal diabetes,the level of IAI was lower(Z= 3. 19, P＜0. 017). No significant difference of insulin sensitivity were observed between infants with and without maternal impaired glucose tolerance (P＞0. 017). The level of HOMA-IR was higher in infants and toddlers with diabetes in first-degree relatives [infants: 3.24(2. 73-4. 13) vs 1.41(0.84-2.50) ; toddlers: 3.98(2.62-4.80) vs 1.70(0.92-3.04); P＜0. 017], while the levels of IAI and FGIR were lower [infants: IAI, -7. 18(-7.42-- 7.00) vs - 6.34 ( - 6.91- 5.82); FGIR, 7.31 (5.40-7.48) vs 14.87 (9.53-25.17); toddlers: IAI,-7.38(-7.57-6.97) vs -6.54(-7.11-5.92); FGIR, 6.17(6.04-8.00) vs 12.65(8.33-21.53), P＜0. 017], and the level of HOMA-β was higher just in the infants [164.00(137.82-198.00) vs 67.06(40.40-106.83), P＜0. 017]. No significant difference was observed in infants with second-degree relatives diabetes and without diabetic family history group (P＞0. 017). Conclusions Maternal diabetes and diabetic family history in first-degree relatives were significantly related to the decrease of insulin sensitivity in infants and toddlers.

Objective To detect the apoptosis of femoral head cartilage cells and to observe the expression of apoptosis-related proteins in the tissues of the femoral head,as well as to explore the main pathway for regulating the apoptosis in steroid induced by juvenile rabbit models with avascular necrosis of femoral head.Methods The models with avascular necrosis of the femoral head and the control group model were made in New Zealand infancy albino rabbits induced by glucocorticoid(GC).The immunohistochemical method was used to detect the expressions of Caspase3,Caspase-8,apoptosis protease activating factor-1 (apaf-1),calpain-1 in the femoral heads.Results 1.The integrated optical density(IOD) values of Caspase-3 in GC-induced subgroup,the subgroup that was not induced by GC and control group were 25 142.72 ± 21 528.48,2 069.63 ± 1 096.96 and 301.80 ± 99.66,respectively.The IOD values of Caspase-8 in GC-induced subgroup,the subgroup respectively and the control group were 24 942.63 ± 18 942.99,2 016.31 ± 1 518.70,236.85 ±97.94,respectively.The IOD values of apaf-1 in GC-induced subgroup,the subgroup that was not induced by GC and the control group were 8 5 14.23 ± 6 384.20,1 118.49 ± 1 360.59,95.13 ± 38.05,respectively.The IOD values of calpain-1 in GC-induccd subgroup,the subgroup that was not induced by GC and control group were 9 636.71 ± 9 123.81,1 881.10 ± 3 277.86,126.71 ± 47.35,respectively.The IOD value differences of the Caspase-3 between GC-induced subgroup and the subgroup that was not induced by GC,the control group were extremely significant (H =l 1.470,23.996,P ＜ 0.01).The IOD value differences of the Caspase-8,apaf-1,calpain-1 between GC-induced subgroup and the control group were extremely significant (H =22.178,22.808,13.553,P ＜ 0.01).2.The linear regression analysis results showed that under the joint action of Caspase-8,apaf-1,calpain-1,only the Caspase-8 could significantly predict Caspase-3,and its regression equation regression got significant effect and could explain 40.3％ of the variance;while the regression effects of the apaf-1 and calpain-1 to Caspase-3 were not significant.Conclusion Death receptor pathway might play a major regulation role in the apoptotic process of avascular necrosis.

A new method for noninvasive prenatal diagnosis of fetal sex was developed by using single-cell PEP-PCR techniques. Micromamipulation techniques were used to obtain single fetal cells from 273 maternal blood samples. The genome of single cells was preamplified by PEP and SRY genes were analyzed by PCR method. The SRY genes of 149 samples were detected by the new method among 153 samples carrying male fetus, while 119 out of 120 samples carrying female fetus were proved negative for SRY genes. The sensitivity and specificity of the new method were 97.39% and 99.17% respectively and the correct rate was 98.17%. The new method has the advantage of high sensitivity and specificity in noninvasive prenatal diagnosis of fetal sex and provides the basis of other researches such as sex-linked inherited diseases.
Adult ; Chromosomes, Human, Y ; Erythroblasts ; chemistry ; Female ; Fetus ; cytology ; Genes, sry ; genetics ; Genetic Diseases, Inborn ; diagnosis ; Humans ; Male ; Maternal-Fetal Exchange ; genetics ; Polymerase Chain Reaction ; methods ; Pregnancy ; blood ; Prenatal Diagnosis ; methods ; Sex Determination Processes

The single cell isolation technique was used to detect fetal nucleated erythroblasts (FNRBCs) at a single cell level from the peripheral blood of pregnant women in order to investigate the feasibility of this method for noninvasive prenatal diagnosis. Single fetal nucleated erythroblasts were isolated from the peripheral blood samples from 51 pregnant women (14 to 26 weeks of gestation) by micromanipulation techniques after density gradient centrifugation. Nested polymerase chain reaction method was used to amplify the SRY gene. It was found that the concordance rate of amplification results with real fetal sex was 82.61 %. The sensitivity and specificity were 80 % and 87.50 % respectively. It was suggested that it is feasible and promising in non invasive prenatal diagnosis to detect fetal nucleated erythroblasts at a single cell level by using micromanipulation techniques.

The single cell isolation technique was used to detect fetal nucleated erythroblasts (FNRBCs) at a single cell level from the peripheral blood of pregnant women in order to investigate the feasibility of this method for noninvasive prenatal diagnosis. Single fetal nucleated erythroblasts were isolated from the peripheral blood samples from 51 pregnant women (14 to 26 weeks of gestation) by micromanipulation techniques after density gradient centrifugation. Nested polymerase chain reaction method was used to amplify the SRY gene. It was found that the concordance rate of amplification results with real fetal sex was 82.61 %. The sensitivity and specificity were 80 % and 87.50 % respectively. It was suggested that it is feasible and promising in non invasive prenatal diagnosis to detect fetal nucleated erythroblasts at a single cell level by using micromanipulation techniques.

Objective: To study the mutation of ENG, ACVRL1, and SMAD4 genes in one of a family of hereditary hemorrhagic telangiectasia (HHT) and explore its molecular pathogenesis. Methods: A family spectrum of a patient with a clinical diagnosis of HHT was surveyed. Peripheral blood samples from proband and their eldest were collected, and ENG, ACVRL1 and SMAD4 gene analysis was performed by chip capture high-throughput sequencing. The mutation detected was verified by Sanger. Results: 9 of the 71 family members were diagnosed with HHT with the main manifestation of recurrent nasal bleeding. Genetic analysis showed that the proband and the eldest son of ENG gene exon 9 frameshift mutation: c.1502-1503insGG (p.Gly501GlyfsX18) , and mutations in ACVRL1 and SMAD4 genes were not detected. Conclusion The frameshift mutation c.1502-1503insGG (p.Gly501GlyfsX18) of the ENG gene is the genetic basis for the pathogenesis of this HHT family.

Objective To compare the effectiveness of expandable intramedullary nail(EIMN) versus locked compression plate (LCP) in the treatment of humeral shaft fractures of AO types A and B.Methods The clinical data were retrospectively analyzed and compared of the 44 humeral shaft fractures which had been treated between May 2012 and February 2016.There were 26 men and 18 women,from 23 to 66 years of age (average,41.5 years).By AO classification,22 cases were type 12-A and 22 type 12-B.EINM was used in 22 patients with an average age of 41.3 ± 1 1.7 years and LCP in 22 ones with an average age of 41.6 ± 10.3 years.The 2 groups were compared in terms of intraoperative blood loss,operative time,hospital stay,union time,union rate,Constant and Mayo scores at the final follow-ups,and complications as well.Results The 2 groups were compatible without significant differences in the preoperative demographic data (P ＞ 0.05).All the 44 patients were followed up for 10 to 18 months (average,12 months).The intraoperative blood loss (76.4 ± 18.66 mL),operative time (69.1 ± 13.2 min),incision length (5.8 ± 1.5 cm) and union time (13.2 ± 8.4 w) in the EIMN group were significantly better than those in the LCP group (138.6 ± 39.4 mL,96.4 ± 14.2 min,8.5 ± 1.4 cm and 18.4 ± 6.6 w,respectively) (P ＜ 0.05).There was also a significant difference between the 2 groups in the total complication rate[18.2％ (4/22) versus 50.0％ (11/22)] (P ＜0.05).No deep infection or should pain was observed in either group.Conclusions Inthe treatment of humeral shaft fractures of AO types 42-A and 42-B,compared with LCP,EIMN may have advantages of less intraoperative blood loss,operative time,union time and complications.The 2 methods are similar in hospital stay and final functional recovery of the should joint.

Objective To investigate the efficacy of anti-infective reconstituted bone xenograft (ARBX) combined with external fixation in the treatment of adult infective nonunion of humeral shaft.Methods A retrospective case series study was conducted to analyze the clinical data of 18 patients with infected nonunion of humeral shaft admitted to Xijing Hospital of Air Force Military Medical University from January 2014 to December 2016.There were 10 males and eight females,aged 19-62 years [(36.9 ± 11.8)years].According to Umiarov classification of infective nonunion,there were 11 patients with type Ⅲ and seven with type Ⅳ.All patients were treated with anti-infective reconstituted bone xenograft (ARBX) combined with external fixation.The number of operations,bone healing time,bone healing rate,infection control rate,postoperative weight bearing time,the time of external fixation removal,postoperative complications,erythrocyte sedimentation rate (ESR),and C-reactive protein (CRP) before and after operation were recorded.Fracture healing and functional recovery were evaluated using the Johner-Wruch lower limb function score.Results The patients were followed up for 12-30 months [(21.3 ±5.6)months].The operation was performed for (1.4 ±0.9) times,with time of bone healing for (16.6 ± 5.8)months,bone healing rate of 83％ (15/18),and infection control rate of 94％ (17/18).The postoperative weight bearing time in 15 patients who obtained bone healing was (3.3 ± 1.5)months after operation,and the external fixation removal time was (18.5 ± 4.2) months after operation.There were three patients with nonunion after operation including one with infection recurrence.Five patients were found with nail tract infection.ESR and CRP at postoperative 3 months [(13.1 ± 8.4)mm/h and (5.6 ± 4.6)mg/L] were significantly lower than those before operation [(47.3 ± 19.2)mm/h and (23.4 ± 7.4) mg/L] (P ＜ 0.05).According to Johner-Wruch lower limb function scores,the results were excellent in nine patients,good in four,fair in one,and poor in four,with excellent and good rate of 72％.Conclusion ARBX combined with External fixation can effectively treat infective nonunion of humeral shaft,improve bone healing rate,and promote function recovery.