Application of droplet digital PCR for non-invasive prenatal diagnosis of single gene disease in two families / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the value of droplet digital PCR (ddPCR) for non-invasive prenatal diagnosis of single gene disease in two families.</p><p><b>METHODS</b>Paternal mutation in cell-free DNA derived from the maternal blood and amniotic fluid DNA was detected by ddPCR. Suspected mutation in the amniotic fluid DNA was verified with Sanger sequencing.</p><p><b>RESULTS</b>The result of ddPCR and Sanger sequencing indicated that the fetuses have carried pathogenic mutations from the paternal side in both families.</p><p><b>CONCLUSION</b>Droplet digital PCR can accurately detect paternal mutation carried by the fetus, and it is sensitive and reliable for analyzing trace samples. This method may be applied for the diagnosis of single gene diseases caused by paternal mutation using peripheral blood sample derived from the mother.</p>

The expression of RPS15a in colorectal cancer tissues and the effects of silencing RPS15a on the proliferation and apoptosis of colon cancer RKO cells / 肿瘤

Objective: To investigate the role of ribosomal protein S15a (RPS15a)in colorectal cancer.Methods: The expressions of RPS15a in 120 specimens of colorectalcancer tissues and the para-cancerous tissues as well as 120specimens of colorectal adenoma tissues were detected byimmunohistochemistry. The proliferation, cell cycle and apoptosisof colon cancer RKO cells after infection with recombinant lentivirusRPS15a-siRNA were detected by Cellomics cell counting assay, colony formation assay and FCM, respectively.Results: The positive rate of RPS15a expression in colorectal cancer tissues was significantlyhigher than those in the colorectal adenoma tissues (86.7% vs 25.8%, P < 0.001) andthe para-cancerous tissues (86.7% vs 11.7%, P < 0.001). The expression of RPS15a wassignificantly correlated with the TNM stage (P = 0.033) and the tumor differentiation (P <0.001) of colorectal cancer. RPS 15a silencing significantly suppressed the proliferation (P <0.01) and colony formation (P < 0.01) of the RKO cells, induced apoptosis (P < 0.01), andarrested the cell cycle at G2/M phase (P < 0.01).Conclusion: The expression of RPS15a in colorectal cancer tissues is higher than those in thecolorectal adenoma tissues and the para-cancerous tissues. Down-regulation of RPS 15aexpression can repress the proliferation and induce the apoptosis of RKO cells.

A novel mutation of GLI3 gene underlying synpolydactyly in a family / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect mutation of GLI3 gene in a family affected with autosomal dominant synpolydactyly.</p><p><b>METHODS</b>Genomic DNA was extracted from peripheral blood samples from members of the family and 100 unrelated healthy controls. Potential mutation was screened by next-generation sequencing and confirmed by Sanger sequencing.</p><p><b>RESULTS</b>A heterozygous frameshift mutation c.480dupC was identified in the GLI3 gene among all patients from the family. The same mutation was not found in unaffected family members and the 100 healthy controls.</p><p><b>CONCLUSION</b>The c.480dupC of the GLI3 gene probably underlies the synpolydactyly in this family.</p>

Analysis of therapeutic outcomes of polycystic ovary syndrome patients with hyperprolactinemia / 中华妇产科杂志

Objective To study effect of drug treatment in polycystic ovary syndrome patients withhyperprolactinemia.Methods We retrospectively studied 63 women with polycystic ovary syndrome and hyperprolactinemia from the Reproductive Medicine Center,Provincial Hospital between January 2005 andMarch 2007.According to the beginning time of bromocriptine,all women were divided into two groups.Group Ⅰ was composed of 48 cases who received bromocriptine administration before induction of ovulation cycles,and the dose of bromocriptine was modulated depending on the level of serum prolactin.When serum prolactin was controlled at normal levels,we decreased the dosage of bromocriptine step by step(1.25 mgonce),and then continued the treatment at maintenance dosage for no less than 3 weeks.After a baselineultrasonographic examination on day 3,patients were treated with clomiphene citrate at a dosage of 100 mg (2 tablets/day)for 5 days of a normal cycle or progesterone-induced bleeding.On day 9,we monitored the growth conditions of follicles routinely with trans-vaginal uhrasound.If there was no dominant follicle,we added human menopausal hormone(hMG,75 U/d)to the protocol.Human chorionic gonadotropin(hCG,6000-10000 IU)was given intramuscularly when the mean diameter of a follicle reached at least 18 mm.At the same time we instructed the patients to have sexual intercourses or carried out artificial inseminationsbefore and after ovulation.Group Ⅱ were 15 cases in which induction of ovulations were commenced almostsimultaneously with beginning of bromocriptine.The same protocol was given to patients in group Ⅱ.The procedures of ovulation induction and the outcomes of treatment were analyzed and compared.Results Compared with groupⅡ.the days of using hMG in Group Ⅰ was shorter by instructing the time of sexualintercourse.The difference was significant(P=0.004).And there were similar results in the artificial insemination cycles(P=0.009).The rate of pregnancy in group Ⅰ(40%,19/48)was higher than that in groupⅡ(27%,4/15),but the difference was not obvious(P=0.525).Conclusion Bromocriptine administration before the stimulated ovulation therapy can decrease the total dosage and treatment course of ovulating drugs.Induction of ovulations simultaneously with start of bromocriptine therapy can shorten the treatment time of infertility.

The mechanism of hyperintense signal at interpeduncle region on diffusion weighted imaging / 中华放射学杂志

Objective To study the mechanism of hyperintensed signal at interpeduncle region in meseneephalon on DWI with diffusion sensitive gradient(Gd)applied perpendicular to the slice direetion (DWIs).Methods MRI scanning was centered at interpeduncle region.and the protocols were as follows:(1)With electrocardiograph(ECG)triggering,DWIs was acquired during systole or diastole periods respectively.(2)To observe the shape of hyperintense signal,DWIs with various slice directions was applied,which was set parallel to skull base,along Y axis,or along the oblique line between Y and Z axis with 30°to Z axis.(3)Fiber tracking was performed with the hyperintense signal as"seed"region,fractional anisotropy(FA)and ADC were derived from(multiple directions diffusion weighted imaging,MDDW).FA and ADC of hyperintease region were compared with those of the nearby fiber by using pair t-tesL Results Signal intensitv of hyperintense region on DWIs were 2296.28±38.19 and 153.81±37.91 for b=0 and b=1000 at systole period.while they were 295.36±38.84 and 154.03±37.52 at diastole period.No staffstical difference wag demonstrated between them(t=1.34,0.62,P＞0.05).The ADC valus on DWIs acquired during systole and diastole periods were(6.07±2.20)×10-4and(6.69±1.44)×10-4 mm2/s respectively.There was rio statistical difference between them(t=0.94,P＞0.05).DTI fiber tracking verified that the hyperintense region located at the decussation of superior eerebelum peduncle(SCP)in mesencephalon.It was in long and narrow heart shape or rectangular shape on DWIs.The shape depended on the direction of Gd The ADC value derived from MDDW at hyperintense region and the fiber below were(10.61±3.42)×10-4 and(9.24±2.21)×10-4 mm2/s respectively.No statistical difference was demonstrated between them(t=0.61.P＞0.05).While,the FA at hyperintensed region 0.43±0.13 was less than that of the fiber below 0.61±0.08(t=8.32.P＜0.05).Conclusions The shape of the hyperintense region on DWIs depends on the direction of Gd.The cardiac period has no effects on the hyperintense signal at peduncles,which may be attributed to the anisotropy of SCP decussation in mesencephalon.

Replacement of androgen receptor gene causes complete androgen insensitivity in a large family / 中华妇产科杂志

Objective To confirm the clinical diagnosis of complete androgen insensitivity syndrome (CAIS) by molecular genetic testing in a large family. Methods PCR was performed to amplify the coding region of androgen gene, followed by direct sequencing in the patients with CAIS and relatives in this family. Results A missense mutation Arg773His was identified in the patients (homozygous) and carriers(heterozygous). Conclusions Mutation Arg773His in the AR gene leads to CAIS in this family. Molecular genetic testing of CAIS facilitates not only prenatal genetic diagnosis but also preimplantation genetic diagnosis and offers genetic counseling for future pregnancies to abandon the transmission of the mutated X chromosome to the coming generation.

Cytogenetic diagnosis on single lymphocyte of DMD patient with exon 50 deletion by two-time duplex PCR / 第三军医大学学报

Objective To perform the cytogenetic diagnosis on single lymphocyte of DMD patient with dystrophin gene exon 50 deletion.Methods Single lymphocytes of a DMD patient with dystrophin gene exon 50 deletion and normal volunteers were picked out and prepared for two-time duplex PCR.Results The rate of precise positive was 92%,91% and 93% in specimens of the patient(SRY positive,exon 50 negative),the male volunteer(SRY positive,exon 50 positive)and the female volunteer(SRY negative,exon 50 positive),respectively.Conclusion Two-time duplex PCR is fit for the genetic diagnosis of single lymphocyte from DMD patient with dystrophin gene exon 50 deletion.

THE UPREGULATIVE EFFECT OF NGF ON THE EXPRESSION OF TNF-? mRNA IN THE LOWER RESPIRATORY TRACT AND VISCEROSENSORY AFFERENT SYSTEM OF THE ASTHMATIC GUINEA PIGS / 解剖学报

0.05).The expression of TNF-? mRNA was significantly upregulated in the lower respiratory tract,C-7-T-5 spinal ganglia and corresponding spinal dorsal horn of the experimental asthmatic guinea pigs compared with the normal saline control group and the simple sensitized group(P

THE EXPRESSION OF NF-?B IN SPINAL GANGLIA OF THE ASTHMATIC GUINEA PIGS AND THE INHIBITORY EFFECT OF DEXAMETHSONE / 解剖学报

Objective To explore the expression of NF-?B in C-7-T-5 spinal ganglia and possible mechanism of treating asthma with dexamethsone. Methods The alterations of NF-?B activity and its mRNA expression were investigated by means of immunohistochemistry and in situ hybridization histochemistry combined with the micro-image analysis in C-7-T-5 spinal ganglia in the asthmatic and dexamethsone-treated guinea pigs. Results The NF-?B immunoreactivity was found mainly in cytoplasm and weakly in nucleus,its mRNA was expressed weakly in C-7-T-5 spinal ganglia in the control groups.The NF-?B immunoreactivity was more in nucleus and less in cytoplasm,the expression of its mRNA increased significantly in C-7-T-5 spinal ganglia in the asthmatic group compared with the control groups(P0.05).Conclusion The present results indicate that NF-?B in C-7-T-5 spinal ganglia might be involved in the pathogenesis of the bronchial asthma,and inhibiting the expression and activity of NF-?B in C-7-T-5 spinal ganglia might be one of the mechanisms of treating the bronchial asthma with dexamethsone.