Objective To pay attention to screening family disease and gene examination in patient with congenital adrenal hyperplasia (CAH). Method In this study, we investigated clinical data and performed gene sequencing of the pedigree and his family members with CAH. Results Based on clinical data, 3 of 6 family members were diagnosed CAH. Bone age of two boys nearly reach the maximum value. Genetic sequencing result indicated a T>A mutation of 1 004 position in CYP21A2 gene, which induced protein change I172N. Conclusion Disease screening and gene examine should be actively executed in patient with CAH in order to obtain diagnosis and start treatment as early as possible.

Objective The aim of this study was to investigate mutations of 41 STR loci. Methods 4546 bloodstain samples were typed from 1932 father–mother–child trios by using AGCU_21+1, AGCU_EX22 and GlobalFiler_ExpressTM amplification Kit. Calculate the mutation rates of STR loci. Results 154 mutations were identified at 32 of the 41 loci. The average mutation rate was 1.0×10-3per locus(95%CI: 0.8~1.1×10-3), and the mutations of SE33 was highest. 152(98.7%) mutation events were one-step mutation, 2(1.3%) events were two-steps. The mutation events occurred in 150 father–mother–child triplets. The mutations in 146(97.3%) triplets occurred at single locus, 8 mutations were observed at two loci in 4(2.7%) triplets simultaneously. 104 paternal and 22 maternal mutations could be determined under 79212 paternal and maternal allelictransfers. The ratio of paternal versus maternal mutations was 4.7:1, and 28 unassigned mutations were observed. Conclusion STR mutation are common in paternity testing, and we should pay more attention to it.

Objective To explore the clinical utilization value of regional blood flow fluorescence visualization in selective arterial clamping robotic partial nephrectomy.Methods 12 cases of robotic partial nephrectomy with indocyanine green-based regional blood flow fluorescence visualization selective arterial clamping between October 2016 and June 2017 by our team were retrospectively analyzed.There were 9 males and 3 females with age between 36-78 years,mean age(51.2 ± 11.0) years,BMI 20.1-36.2 kg/m2,mean of (25.6 ± 4.8) kg/m2,tumor diameter 2.0-5.1 cm,mean of(3.3 ± 0.9) cm,and R.E.N.A.L.score 4-10,mean(7.3 ± 2.0).Preoperative renal function status of eGFR (estimated glomerular filtration rate) was 82-133 ml/(min · 1.73 m2),mean (101.9 ± 13.7)ml/(min · 1.73 m2) and split ECT-GFR of 44.5-70.6 ml/min,mean of(53.8 ± 8.5) ml/min in operated kidney;48.2-71.1 ml/min,mean of(56.8 ±6.8) ml/min in contralateral kidney;in total,92.7-139.1 ml/min,mean of(109.8 ± 14.6)ml/min.Perioperative information including operative time,blood loss,warm ischemia time,surgical complications,pathologic outcomes and follow-up data of included patients were analyzed.Results All procedures were done smoothly without open or radical conversion,with operative time of 95-203 min,mean of (170.6 ±38.6)min,and estimated blood loss of 60-1 000 ml,mean of(178.3 ± 206.9)ml.According to the visualize uptake of fluorescence imaging perfusion area visualization after selective arterial clamping,1 case underwent unclamping robotic partial nephrectomy,1 case converted to main renal artery clamping with warm ischemia time of 18 min,10 cases performed via renal arterial branch clamping with an average ischemia time of (25.5 ± 10.5) (range 17-46)min,1 of which due to parenchymal bleeding obscuring visualization necessitated clamping of the main renal artery with 1000 ml blood loss,46 min of arterial branch occlusion and 16 min of main artery clamping.The average postoperative hospital stay was (5.8 ± 0.9) (range 5-8)days,and the duration of drainage was (3.5 ± 0.5) (range 3-4) days.No postoperative complications occurred.Postoperative pathology:all margins were all negative,11 cases of clear cell carcinoma,eosinophilic adenoma in 1 case.The average eGFR of 9 cases of renal arterial branch clamping was (94.5 ±22.5)(range 56-140)ml/(min · 1.73 m2) at discharge with a 5％ percentage decrease versus preoperative level.Among them,4 cases obtained with ECT-GFR data 1 month postoperatively had a mean of(37.6 ±13.2)(range 20.8-55.8) ml/min with 29％ percentage decrease on the surgical side,an average of (58.5 ± 6.9) (range 51.2-68.4) ml/min with 2％ compensatory increase of the contralateral side,and (98.7 ± 16.2) (range 79.3-124.3) ml/min in total with a 10.4％ overall decrease.Conclusions The visualization of blood flow imaging based on indocyanine green fluorescence can clearly and intuitively show the effect of branching arterial occlusion in robotic partial nephrectomy,guide the optimization of surgical resection strategy,and improve safety and clinical outcome.

The application of Problem-based Learning(PBL) in prosthodontics accommodates the request of teaching reformation and also elevates the quality of students and teachers.PBL is a fine teaching method and deserves popularization.

Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.

Objective To explore the causes of biliary complications related with liver donor fol- lowing liver transplantation.Methods Ninty-nine patients with improved liver donor treatment during liver transplantation from May 2005 to April 2006 were followed up and the clinical data were ana- lyzed.At the same time,the rate of biliary complications was compared with that occurring on 43 pa- tients with unimproved liver donor treatment.Results Only 4 in 99 patients with improved liver donor treatment had biliary leakage with the rate of biliary complications being 4% in comparison with 11% in those with unimproved liver donor treatment.Conclusion The improvement of liver donor treat- ment,including shortening heat-ischemia time,completely washing bile duct and remaining the whole blood supply of bile duct,can decline the occurrence of biliary complications.

This study is to establish the fingerprint for Phyllanthus emblica and their tannin parts from different habitats by HPLC for its quality control. The determination was carried out on a Diamonsil C18 (4.6 mm x 250 mm, 5 microm) column, with methanol-0.2% glacial acetic acid as mobile phase with gradient elution at a flow rate of 1 mL x min(-1). The temperature was maintained at 30 degrees C and the detected wavelength is 260 nm, Thirteen chromatographic peaks were extracted as the common peaks of the fingerprint of P. emblica, and eleven as the common peaks of P. emblica tannin parts, and five peaks were identified by comparing with referent samples. The fingerprints of 8 samples were compared and classified by similarity evaluation, cluster analysis and principal component analysis (PCA). The similarity degrees of eight P. emblica were between 0.763 and 0.993, while tannin parts were between 0.903 and 0.991. All the samples of P. emblica and their tannin parts were classified into 3 categories. The method was so highly reproducible, simple and reliable that it could provide basis for quality control and evaluation of P. emblica from different habitats.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Medicine, Tibetan Traditional ; Phyllanthus emblica ; chemistry ; classification ; Quality Control ; Tannins ; analysis ; Tibet

AIMTo apply stochastic resonance algorithm (SRA) to quantitative analysis of weak chromatographic signal, which was embedded in the noise.

METHODSBased on the theory of stochastic resonance (SR), a simple and effective SRA has been established to improve analytical detection limits of chromatographic analysis, which apply to enhance the signal to noise ratio by the optimization of the parameters and Runge-Kutta method, was established. The method was used to quantitative analysis of phenazopyridine in human plasma by HPLC/UV. Meanwhile this method is compared with HPLC/MS.

RESULTSBy experimental chromatographic data sets, an excellent quantitative relationship between concentrations of phenazopyridine and their responses had been obtained. The concentration of phenazopyridine in plasma determined by HPLC/UV with SRA and HPLC/MS showed that there was no significant difference (P > 0.05) between the two methods.

CONCLUSIONThe new method was feasible.

Chromatography, High Pressure Liquid ; methods ; Humans ; Mass Spectrometry ; Noise ; Phenazopyridine ; blood ; Sensory Thresholds ; Spectrophotometry, Ultraviolet ; Stochastic Processes

Objective To investigate the clinical effect of porcelain fused for metal crown restoration after root canal therapy.Methods Totally 90 patients with pulpitis or periapical periodontitis from December 2013 to December 2015 had their clinical data analyzed,who underwent root canal therapy and were divided equally into an observation group and a control group.The observation group took porcelain endocrowns repair and the control group applied ceramic crown.The two groups were compared on the restoration integrity,edge sealing,residual rate,gum,food impaction and color 3 months,6 months and 1 a after treatment.Results The observation group behaved significantly better than the control group in the edge sealing and restoration integrity (P＜0.05).There were significant differences between the residual rates,gum and food impaction in the two groups (P＜0.05).Conclusion Porcelain endocrowns repair gains advantages over ceramic crown repair in edge sealing,integrity,gum,food impaction and color after root canal therapy,though its long-term effect needs further investigation.

Objective To investigate the immunogenicity of immunodominant cytotoxic T lymphocyte epitope E749-57 of human papilloma virus (HPV) 16 oncoprotein E7 chaperoned by heat shock protein (HSP)110. Methods Mouse HSP110 gene was cloned into prokaryotic expression vector pQE-80L for the expression of HSP110 protein, which was purified using Ni-NTA column. SDS-PAGE and Western-blot were conducted to confirm the purified mHSP110 protein, which was subsequently incubated with E749-57 peptide under heat shock condition, and high-performance liquid chromatography (HPLC) was used to evaluate the binding efficiency of the recombinant protein and E749-57 peptide. Twenty mice were divided into 4 groups to be immunized with mHSP110 protein, E749-57 peptide, mHSP110-E749-57 complex and phosphate buffered saline (PBS),respectively. Two weeks after the last immunization, spleen cells were collected from the immunized mice and divided into 2 parts: one were stimulated by E749-57 peptide followed by the detection of CD8+ INF-γ+ T cells with flow cytometry; the other one were subjected to MTT analysis for the estimation of cell proliferation. The mHSP110-E749-57 complex was also used to immunize TC-1 tumor bearing mice to observe its anti-tumor effect.Results The full-length 2577 bp-sized mHSP110 gene was amplified from mouse liver cDNA and cloned into pQE-80L vector. Direct sequencing confirmed the correctness of the cloning. SDS-PAGE and Western-blot demonstrated the successful purification of mHSP110. HPLC assay showed that the purified mHSP110 protein could bind with E749-57 to form a relatively stable protein complex. The percentage of IFN-γ+ CD8+ T cells in and proliferation index of spleen cells from the complex-immunized mice were statistically higher than those from the other 3 groups of mice. Moreover, the complex could obviously inhibit the growth of TC-1 tumor in mice. Conclusion The mHSP110-E749-57 complex could enhance the generation of specific cytotoxic T lymphocytes and exert anti-tumor effects in mice.