Due to a wide variety of classification and phenotypes,rare diseases are difficult to be diagnosed in the clinical practice Studies have shown that most rare diseases belong to inherited diseases,so it is particularly important to carry out the genetics research and molecular diagnosis.Next generation sequencing (NGS) technology has the advantages of a high throughput,high sensitivity etc,which is the main research means to identify the pathogenic genes of rare diseases at present.With the development of NGS and the bioinformatics technology,in recent years the whole exome sequencing and targeted panel sequencing have been gradually applied to clinical molecular diagnosis,which is important to increase the accuracy of diagnosis and to improve the therapeutic effect of rare diseases.

The lumbar part of the sympathetic trunk in 206 sides of cadavers was observed. The results were outlined as follows:1. The average length of the lumbar sympathetic trunk in the adults is 17.58?4.51 cm on the left side and 16.60?5.14 cm on the right side respectively.2. In most cases (40.29?3.42%), there are 4 lumbar sympathetic ganglia on each side. They may fuse with each other between themselves, or the 1st lumbar ganglion fuses with the lowest thoracic ganglion (12.60?2.31%), or the lowest lumbar ganglion fuses with the 1st sacral ganglion (6.31?1.69%).3. The lumbar sympathetic ganglion gives off ramus communicans connecting the corresponding spinal nerve. The number of the ramus communicans of each ganglion varies from 0 to 6, but in most cases, 2～3, and decreases from above downwards along the trunk. There are frequently two or more rami in the upper lumbar ganglia, but only one in the lower ganglia.4. The number and sites of the lumbar ganglia on both sides of an individual are rarely symmetric (1.98?1.37%). One of these ganglia is situated constantly near the brim of pelvis or the common iliac artery, and accordingly it can be found easily.Most of the lumbar arteries are situated deep to the lumbar sympathetic trunk, only in a few cases, they are situated superficially, and all of them occur on the left side. In 25.75?3.05%, the lumbar veins are likewise located superficially. This can be found in any pair of the veins, but frequently at 3rd or 4th ones.The observation may be helpful to surgical operation.

The intertubercular grooves of 212 humerus were studied. The statistical results showed that the depth of intertubercular grooves is highly correlated with the slope of the medial wall of the groove (r=0.116, p=0.05, lineal regression: =50.01+9.04x). It means that the deeper the groove the bigger the slope of the medial wall is, and the shallower the smaller. Their functional relationship has practical significance. The normal surface contour of the intertubercular groove is the important condition that keeps the long head tendon working normally. Therefore internal or external causes, e. g. fracture of the surgical neck, hyperplasia of the bone in aged people, may alter the normal anatomical relation of the intertubercular groove, which precipitated to tendosynovitis of the biceps. By observation of the compactness of the surgical neck cortex in three different levels, we found that it is in a transitional zone of from thin to thick, which agrees with the explanation of why fracture occurs frequently at the surgical neck.

Objective To study the influence of the preparation and quality control of Xiaochuan cataplasm on its drug action. Methods Xiaochuan cataplasm was prepared with hydrosoluble polymers matrix. A sequence of tests such as adhesive force test, recontour test, paste-content test, cold-resistance test, heat-resistance test, toxicity and pharmacodynamic action tests were performed on this drug. Results Xiaochuan cataplasm contains a sound volume of medicine, and shows good adhesive property, no sensibilization and stimulation, and notable drug action. Conclusion The results demonstrate that the preparation craft of Xiao-chuan cataplasm is reasonable and practicable., the quality is controllable and the method is a safe and effective transdermal drug delivery system.

Objective To study the clinical characteristics and iduronate-2-sulfatase (IDS)gene mutation of one child patient with mucopolysaccharidosis type Ⅱ (MPS Ⅱ).Methods All the 9 exons of IDS gene were amplified by polymerase chain reaction (PCR)technlogy.The PCR products were screened by direct gene Sanger sequencing.Results A missense muta-tion (c.445T>C)on exon 4 was found after the analysis of the gene sequencing results of PCR products in this patient’s IDS gene.Thi smutation leaded to the 149th codon TCT encoded serine into a CCT encoding proline (p.Ser149Pro).Mean-while,the IDS gene in the parents were widetpye,so this was a de novo mutation.Conclusion The de novo mutation of IDS gene is the cause of our patient with?mucopolysaccharidosis,one novel mutation (p.Ser149Pro)was identified.

ObjectiveTo establish an effective method for F9 gene mutation detection by using high resolution melting ( HRM ) curve analysis.Methods Peripheral blood samples of 55 hemophilia B (HB) patients were collected from Shanghai Ruijin Hospital during January 2005 to June 2010.Genomic DNA was extracted from the peripheral blood.PCR amplification combined with sequencing was used to identify the F9 gene mutations in 40 patients.HRM assay was established on the 21 DNA samples with known mutations in exonl to exon7 of F9 gene.Mutation scanning of exonl to exon7 by HRM combined with direct sequencing of exon8 was used in the molecular diagnosis of 15 HB patients with unknown F9 gene mutations.ResultsF9 gene mutation was detected in each of the 40 HB patients by direct sequencing.By HRM,the different melting curve patterns were identified in 19 out of 21 cases.The detection rate was about 90％.Through mutation scanning of exonl to exon7 by HRM combined with direct sequencing of exon8,F9 gene mutations were detected in all the 15 HB patients with unknown F9 gene mutations.Thirty-four F9 gene mutations had been identified in the 55 HB patients.ConclusionsA new strategy of HB genetic diagnosis,scanning mutations of exonl to exon7 combined with DNA sequencing of exon8 of F9 gene,is established in this study.The new strategy is efficient and reliable.

AIM: This research emphasizes the effect of ganglioside GM1 on generation of the intracellular amyloid ?-protein(A?) and investigates where and how GM1 promoted the production of intracellular A?, particularly the more highly amyloidogenic A? 42 which is basis of senile plaque.METHODS: Human neuroblastoma cells transfected with human amyloid precursor protein (APP) cDNA were used to analyze the effect of the various concentrations of GM1 on the level of intracellular total A? by IP-Western blot. Subcellular compartment localized and colocalized with intracellular A? 42 was determined by double or triple immunofluorescence labeling.RESULTS: The intracellular total A? was promoted by GM1, and the levels of intracellular A? were correlated to the concentrations of GM1 in a dose-dependent fashion ( P

Objective To evaluate the value of PCR-restriction fragment length polymorphism (PCR-RFLP),real-time PCR and multiplex ligation-dependent probe amplification (MLPA) in the genetic diagnosis of spinal muscular atrophy (SMA) and make laboratory support accessible to clinicians for the molecular diagnosis of SMA.Methods Methodological evaluation.Forty-one suspected SMA cases and 359 control individuals received in Shanghai Children's Medical Centre from March 2013 to June 2014 were detected for the deletion of exon 7 and 8 in the survival motor neuron gene 1 (SMN 1) by PCR-RFLP,realtime PCR and MLPA,respectively.Then the results of the three methods were compared and the benefits and limitations of the three methods were evaluated.Results The result of real-time PCR was in complete agreement with that of MLPA,showing that 29 suspected cases harbored homozygous deletions of SMN1 and 1 case possessed heterozygous deletion.Among the homozygous deletions,27 patients demonstrated absence of exon 7 and 8,and 2 cases demonstrated only the absence of exon 7.Meanwhile,both PCR-RFLP and MLPA analysis showed the same results that only 5 out of 395 control cases carried heterozygous deletion.As for cases without heterozygous deletions,PCR-RFLP demonstrated the same result with real-time PCR and MLPA but it missed all the heterozygous ones.Conclusions PCR-RFLP,the conventional SMA gene diagnosis method,was only capable of detecting homozygous deletion of exon 7 and/or 8 of SMN1,but was not as sensitive as to find out the carriers with heterozygous deletions.MLPA could detect the deletion and quantify the copy numbers of exon 7 and 8 of SMN1,efficiently,while the price was relatively high,which brings challenges for its application in the carrier screening of SMA.Compared with these two methods,realtime PCR with high throughput and low input was a rapid,acourate and economic method for the genetic diagnosis of SMA and carrier screening in large populations.

Objective To discuss the clinical application of multi-slice spiral CT angiography (MSCTA) in acute spontaneous intracranial hemorrhage .Methods 41 cases of acute spontaneous intracranial hemorrhage diagnosed with CT (including subarachniod cavity hemorrhage 29, intra-cerebral hematomas 12)were examined with MSCTA,the techniques of volume rendring (VR) and maximum intensity projection(MIP)were used. 7 cases of intracranial aneurysm closed by titanic clamp , 2 cases of cerebral arteriovenous malformation(AVM) and one case of AVM treated by intra-zest aneurysm excision,the post-operative MSCTA examination were received.Results 11 cases of intra-cerebral aneurysm, 4 cases of cerebral AVM and 1 case of AVM with intra-zest aneurysm were found among the 41 cases of acute spontaneous intracranial hemorrhage. MSCTA clearly demonstrated the size,neck and artery of aneurysms as well as the location, size ,zest shape, artery and vein of AVM. Pre-operative MSCTA findings of 7 cases of intracranial aneurysm clapped by titanic clap ,2 cases of cerebral AVM and 1 case of AVM treated by intra-zest aneurysm excision were confirmed with the operative findings.Post-operative MSCTA showed that the clamp location was normal; arteries were passable; zests of AVM were excised. Conclusion MSCTA is a non-injured, rapid and effective way to find the cause of acute spontaneous intracranial hemorrhage.It also has clinical application of post-operatiove assessment for intracranial aneurysm and cerebral AVM.

Objective To investigate the changes of serum trace elements and nutritional proteins in children with acute lymphoblastic leukemia and acute myeloid leukemia at the stage of remission.Methods Erythrocyte count,hemoglobin,serum levels of total protein,albumin,iron,ferritin,transferrin,lactate dehydrogenase,ceruloplasmin,cuprum,zinc and their ratio were measured in 43 patients with acute lymphoblastic leukemia,19patients with acute myeloid leukemia at stages of remission(remission groups),and 30 healthy controls(control group)enrolled from Shanghai Children's Medical Center using atomic absorption spectrometry,nophelometry assay,dry chemical method,and chemiluminescence method.The differences of these indicators between remission groups and control group were analyzed.Results Serum levels of total protein(P=0.454),iron(P=0.769),transferrin(P=0.903),and zinc(P=0.343)were not significantly different between the remission groups and the control group.Serum levels of ferritin(P=0.000),lactate dehydrogenase(P=0.000),ceruloplasmin(P=0.000),cuprum(P=0.002),and Cu/Zn ratio(P=0.003)in the remission groups were significantly higher than those in control group.On the contrary,erythrocyte count(P=0.000),hemoglobin(P=0.000)and albumin(P=0.046)were significantly lowerin remission groups than those of control group.Serum levels of all detected indicators were not significantly different between the acute lymphoblastic leukemia remission group and acute myeloid leukemia remission group(P>0.05)except for lactate dehydrogenase(P=0.025).Conclusion At the remission stage of acute lymphoblastic leukemia and acute myeloid leukemia,serum levels of some trace elements and nutritional proteins gradually returned to normal,and the original balance is established again.