BACKGROUND:Hydroxyapatite-coated titanium substrates improve the biocompatibility and have the property of intimating ossteointegration with host bonebed. However, hydroxyapatite lacks the necessary mechanical strength and degrades easily in the extracelular fluids, which may affect the stability of the titanium implant.
OBJECTIVE:To study the synthesis and characterization of lanthanum-incorporated hydroxyapatite coatings.
METHODS: Lanthanum-incorporated hydroxyapatite coatings were prepared by hydroxyapatite and 10%, 20% and 30% lanthanum, respectively, by means of sol-gel, which were then deposited on titanium substrates with dip-withdrawal technique. Surface morphology and crystaline microstructure of the coatings were observed by scanning electron microscope. The presence of functional groups for the obtained samples was performed by Fourier transform infrared absorption spectroscopy and X-ray diffraction. The Ca2+ concentration released from the coatings was measured by atomic absorption spectrometry for analysis of degradation property.
RESULTS AND CONCLUSION: With the increase of lanthanum content, the diffraction peak and crystalinity of lanthanum-incorporated hydroxyapatite coatings were increased, but the whole structure of lanthanum-incorporated hydroxyapatites had little changes. The crystal structure maintained stable with charge balance. The lanthanum-incorporated hydroxyapatite coatings showed uniform and high-dense structure and were free of cracks, indicating the coatings had good bonding strength. Under the simulated biological environment, based on the determination of Ca2+ release from the coatings, we can conclude the lanthanum-incorporated hydroxyapatite coatings have a stronger acidoresistance.

Objective To establish a TaqMan-MGB fluorescent probe characterized real-time polymerase chain reaction ( qPCR) method for detecting retinoic acid induced genes G ( RIG-G) in human acute promyelocytic leukemia ( M3 ) .Analyze RIG-G expression levels in peripheral blood of both normal persons and M3 patients and explore its diagnosis value for M 3.Methods Methodology establishment study.A detection method and standard curve of TaqMan-MGB real-time PCR were established after designing specific primers and TaqMan-MGB fluorescence probe of human RIG-G gene and using reverse transcription complementary DNA ( cDNA) as a template.The performance of this method was evaluated in specificity, accuracy, precision, analytical sensitivity and interference substances . Twenty clinical specimens with M3 were quantified RIG-G expression so as to evaluate the correlation between peripheral blood and bone marrow samples .Meanwhile , the results of RIG-G expression in peripheral blood of 40 normal specimens and 20 patients with M3 were analyzed by t-test.And receiver-operating characteristic curve ( ROC ) was used to analyze the detection efficiency of M 3.Results There was a good linear relationship between log value of RIG-G standard substance and threshold cycle number ( Ct ) ( standard curve equation:Y=-3.539X+42.952,R2 =0.999).New method was used to detect standard substance . The deviation between observed and expected values was <5% (r=0.999).Three concentration samples (107 ,104 ,101 copies/μl) were selected for precision test.Intra-assay coefficients of variation were 1.38%, 2.31% and 1.38%, respectively , and intre-assay coefficients of variation were 0.71%, 1.17% and 5.07%, separately.All were less than 10%.The sensitivity of this method was 101 copies/μl.There was a good correlation of RIG-G results between peripheral blood and bone marrow in M 3 patients(r=0.996, b=0.973).But there was no significant difference between this two group results (t=0.099, P>0.05). However , there was obvious difference of RIG-G value in peripheral blood between control group and M 3 patient group (U=18,P<0.001), 3.62 ×104(1.61 ×104 -4.90 ×104)copies/μl for controls and 7.10 ×102 (5.43 ×102 -2.21 ×103 ) copies/μl for M3 patients, respectively.Conclusions Successfully establishe a TaqMan-MGB real-time PCR method for detecting RIG-G gene in peripheral blood.The accuracy, precision, sensitivity and specificity are good .It could provide necessary help in early diagnosis and monitor treatment of clinical M3 patients.

Background and purpose:Retinoic acid-induced gene G (RIG-G) is a tumor suppressor gene which is cloned by NB4 cell line from a acute promyelocytic leukemia cell. This study aimed to investigate the effect ofRIG-G in lung cancer cells A549 by constructing a lentiviral vector expressing RIG-G under doxycycline (DOX) regulation.Methods:RIG-G gene ampliifcation was performed by quantitative real-time PCR (qRT-PCR). pLenti6/TO/V5-GIM-RIG-G lentiviral vector withGFP was built by LR recombination system. The concentration of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were measured by virus titer method. After infecting A549 cells, stably transfected lines were selected via limiting dilution analysis.RIG-G gene expression was examined by immunolfuorescence staining and Western blot assay. Cellular proliferation was determined by CCK-8 assay.Results:The concentrations of pLenti6/TO/V5-GIM-RIG-G lentiviral vector andTet-on lentiviral vector were 1.0×108TU/mL and 4×109 VP/mL, respectively. RIG-G was expressed in lentivirus infected A549 cells after adding DOX, and the amount of cells withGFP could be observed by lfuorescence microscopy.After the expression of RIG-G protein, the prolif-eration activity of A594 cell was signiifcantly inhibited compared to the control group (1.168±0.107vs 2.099±0.162, P<0.05).Conclusion:The regulated expression ofRIG-G gene was established in A549 lung cancer cell line. The RIG-G protein has potential abilities to inhibit the proliferation of lung cancer cell A549.

Objective To evaluate the serum level of antimicrobial peptide human cationic antimicrobial protein 18 ( hCAP18 ) in non-small cell lung cancer ( NSCLC ) patients and its auxiliary diagnosis and prognosis value.Methods Case-control study was used.The serum level of hCAP18 was measured by enzyme linked immunosorbent assay ( ELISA) in 50 cases with NSCLC patients of department of thoracic surgery and 50 cases healthy people of department of physical examination from January 2011 to January 2012 in Tongji Hospital of Tongji University.The concentrations of hCAP18 in serum of NSCLC patients before and after surgery were analyzed.The sensitivity and specificity of serum hCAP18 for the diagnosis of NSCLC were evaluated using the receiver operating characteristic ( ROC ) curves.Data was analyzed by using the t-test and Log-rank test.Results Serum hCAP18 concentration in NSCLC patients (6 733 ±771.8) μg/L was significantly higher than in healthy controls (253 ±6.9) μg/L (t=8.396, P<0.05) .However, the concentration of hCAP18 showed no significant difference between squamous cell carcinoma and adenocarcinoma[(6 300.0 ±1 221.0) μg/L and (7 074.0 ±1 005.0) μg/L, respectively;t=0.494 2, P <0.05 ] .hCAP18 levels had significantly decreased in serum of NSCLC patients after 30 d surgery compared to preoperative results[from (6 733.0 ±771.8) μg/L to (433.6 ±38.2)μg/L;t=8.512, P<0.05].ROC analysis of serum hCAP18 yielded an AUC (Area under the ROC curve) of 0.931 ( 95% CI =0.884 -0.978 ) with 95% sensitivity and 96.3% specificity, which was higher than the CYFRA21-1[0.873 (95%CI=0.758-0.917)].The relapse rate of NSCLC patients with serum hCAP18≤390.0 μg/L was 12.5%(4/32), while 44.4%(8/18) in NSCLC patients with serum hCAP18>390.0μg/L (χ2 =22.64,P<0.05).Conclusions Detection of serum hCAP18 shows a good sensitivity and specificity for the auxiliary diagnosis of NSCLC. It is possible to be a potential detection index for noninvasive diagnosis and monitoring progression of lung cancer.

Objectve To evaluate the serum level of antimicrobial peptide human cationic antimicrobial protein 18 (hCAP18) in colorectal patients and it auxiliary diagnosis and prognosis value.Methods Case-control study was used.The serum level of hCAP18 was measured by enzyme linked immunosorbent assay(ELISA) in 68 cases with colorectal patients of department of gastrointestinal surgery and 40 cases healthy people of department of physical examination from January 2014 to Junc 2015 in Tongji Hosptial of Tongji University.The concentrations of hCAP18 in serum of colorectal patients before and surgery were analyzed.Immunohistochemistry was used to detect hCAP18 expression in colorectal carcinoma.The effect of hCAP18 on colon carcinoma cell proliferation was detected by BrdU-ELISA and soft agar colony formation assay.The sensitivity and specificity of serum hCAP18 for the diagnosis of eolorectal were evaluated using the receiver operating characteristic curves(ROC).Date was analyzed by using the ttest and one-way analysis of variance.Results hCAP18 serum levels in colon cancer of stage Ⅰ,Ⅱ,llⅢ and Ⅳ patients were (0.46 ± 0.18) mg/L,(0.65 ± 0.45) mg/L,(1.26 ± 0.68) mg/L and (2.35 ± 1.06)mg/L.Mean value was(1.16 ±0.88) mg/L,which was significantly higher than in normal people (0.19 ±0.07) mg/L (t =5.290,P ＜ 0.05).hCAP18 levels had significantly decreased in serum of colorectal patients after 30 d surgery compared to preoperative results [from (1.16 ± 0.88) mg/L to (0.26 ± 0.06) mg/L;t =3.971,P ＜ 0.05].Immunohistochemistry results showed hCAP18 was high expression in colon cancer tissue compared with adjacent tissues;BrdU-ELISA assay results showed HCTll6 and SW480 cell proliferation increased significantly after 0.05-1 mg/L of hCAP18 treatment;Soft agar clone formation experiment proved hCAP18 could significant enhance clone formation of HCT116 and SW480 colon cancer cell lines.The size of clonal cluster of HCT116 was increased from (145.40 ± 35.20) μm to (370.80 ± 32.65) μm (t =10.50,P ＜ 0.05) and SW480 was increased from (101.00 ± 27.10) μm to (369.00 ± 27.29) μm (t =15.58,P ＜0.05);The numbers of clonal cluster of HCT116 was increased from 8.50 ± 2.30 to 42.80 ± 6.60 (t =3.945,P ＜ 0.05) and SW480 was increased from 6.20 ± 1.70 to 46.00 ± 7.20 (t =4.775,P ＜ 0.05).ROC analysis of serum hCAP18 yielded an AUC (area under the ROC curve) of 0.93 (95％ CI =0.859-0.999)with 91.17％ sensitivity and 80.00％ specificity,which was higher than the CEA[0.78 (95％ CI =0.699-0.933)].Conclousions Detection of serum hCAP18 shows a good sensitivity and specificity for the auxiliary diagnosis of colon cancer.It is possible to be potential detection index for noninvasive diagnosis and monitoring progression of colon cancer.hCAP18 could promote the proliferation of colon cancer cells,it played an important role in the progression of colon cancer.

HBV reactivation is commonly seen during immunosuppressive therapy and is associated with high incidence and mortality rates due to hepatitis outbreak and liver decompensation, and therefore, it should be taken seriously. However, the prevention and management of this potential complication is still a difficulty in clinical practice. This article reviews the diagnostic criteria and clinical outcomes of HBV reactivation, discusses the association of immunosuppressive therapy with the risk of HBV reactivation, and outlines the strategies for the prevention of HBV reactivation and recent advances. It is pointed out that early identification of patients with HBV infection before immunosuppressive therapy is of vital importance, and the initiation of antiviral therapy at the right moment based on risk stratification can effectively reduce the risk of HBV reactivation. We hope that this review can increase the awareness of HBV reactivation among clinicians and provide an effective reference for optimizing the management and prevention of HBV infection.

Objective:To investigate the clinical and molecular genetic characteristics of 6q24-related transient neonatal diabetes mellitus (6q24-TNDM).Methods:The clinical data of two neonates with 6q24-TNDM admitted to Shanghai Children's Hospital, Shanghai Jiao Tong University in 2017, were retrospectively collected. The methylation levels of 16 cytidine-phosphate-guanosine (CpG) sites from the methylated differentially modified region (DMR) in 6q24 were quantitatively analyzed by pyrosequencing.Results:Case 1, aged 5 d, was born at 37 +4 gestational weeks due to fetal growth restriction, and case 2 was 11-days old and born at 38 +2 gestational weeks. Both infants were male and small for age. They were born through a cesarean section. The birth weight of case 1 and case 2 were 2 340 g and 2 600 g, respectively. They were admitted due to hyperglycemia with blood glucose of 12.95 and 8.00 mmol/L on admission, respectively. Physical examination showed slightly poor skin elasticity and thin subcutaneous fat. Laboratory examination revealed lower serum insulin (<1.39 and 3.94 pmol/L) and peptide C (0.05 and 0.14 nmol/L) levels, positive results of urine glucose, negative tests for urine ketone, serum anti-glutamic acid decarboxylase antibody, anti-insulin antibody, and islet cell antibody in both cases. Normal size of the pancreas was observed by ultrasonography. The infants were improved and were discharged after subcutaneous insulin infusion for more than two weeks. The treatment was discontinued at 69 d and 42 d postnatally for case 1 and case 2. Prenatal diagnosis of the two infants showed normal karyotypes and uniparental disomy of chromosome 6 indicated by single nucleotide polymorphism chip. No pathogenic mutations were detected by next-generation sequencing after admission. The methylation levels of 16 CpG sites in DMR of 6q24 in the two cases, which were quantitatively analyzed by pyrosequencing, were lower than 10% (normal value in healthy matched controls: 40%), indicating an obvious hypomethylation. Conclusions:For children with TNDM who are small for gestational age at birth, presenting hyperglycemia with decreased serum insulin and C-peptide levels, pyrosequencing can be used to quantitatively analyze the methylation levels of CpG sites in 6q24 DMR, which can quickly and directly assist in the diagnosis of 6q24-TNDM, thereby contributing to the treatment and prognosis assessment.

Objective To verify the performance of a novel HBV DNA assay based on AUTRAX automatic nucleic acid extrac-tion workstation .Methods According to the evaluation protocols of Clinical and Laboratory Standards Institute (CLSI) ,the per-formance of a novel HBV DNA assay was assessed in the aspects of accuracy ,precision ,lower limit detection ,linearity ,interference rejection and pollution prevention .Results The accuracy rate of this assay for laboratory evaluation samples of Shanghai Clinical Laboratory Center was 100% during 2015 to 2016 ,with each deviation between observed and expected values within ± 0 .18 lg IU/mL .The intra-assay and inter -assay CV of two levels (106 ,104 ) samples were both less than 5% .Detectable rates of lower limit of detection (LOD) and lower limit of quantitation (LOQ) were 5/5 at 20IU/mL and 40 IU/mL ,respectively .And intra-assay CV of LOQ was 3 .18% ,less than 5% .Linearity assessment exhibited an excellent dynamic range of linear quantification from 40 to 108 IU/mL(Y=1 .0182X-0 .3182 ,R2 =0 .978) .According to product manual ,conjugated bilirubin ,hemoglobin and triglyceride as interfering substance were made at concentration of 600 μmol/L ,7 g/L and 4 .5 mmol/L ,separately ,which had no interference for two levels (106 ,104 ) samples .Each deviation value between interference group and control group was within ± 0 .45 lg IU/mL .No pollution phenomenon was found .Conclusion The novel HBV DNA quantification by AUTRAX automatic nucleic acid extraction workstation has excellent performance in aspects of accuracy ,precision ,lower limit detection ,linearity ,interference rejection and pollution prevention ,which can be used for the detection of clinical specimens .

Objective:To explore the role of infliximab (IFX) in the repairation of intestinal mucosal barrier in Crohn′s disease (CD).Methods:From January 2018 to October 2019, in Shanghai Tenth People′s Hospital, 382 CD patients were selected. All the patients were treated with IFX. And 103 individuals who underwent colonoscopy were selected as healthy control group. The general clinical data, fasting blood samples and intestinal mucosa tissue samples of CD patients and healthy controls were collected. The body mass index (BMI), hemoglobin, albumin, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and relative inflammation factors, including tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-1, IL-2, IL-6, IL-8, IL-10 and IL-17A, and their mRNA expression levels were detected. Crohn′s disease activity index (CDAI) and simplified endoscopic score for Crohn′s disease (SES-CD) were used to evaluate the disease activity of CD patients. The expression levels of occudin, claudin-1, zonula occluden-1 (ZO-1) and junctional adhesion molecule-A (JAM-A) were measured by Western blotting. The intestinal mucosal epithelial cells were observed by transmission electron microscope. T test was used for statistical analysis. Results:Before treatment, BMI, and hemoglobin and albumin levels of CD patients were all lower than those of healthy control group ((18.3±1.8) kg/m 2 vs. (20.2±1.2) kg/m 2, (95.3±8.4) g/L vs. (129.2±5.7) g/L, (33.2±5.4) g/L vs. (50.3±3.2) g/L), and the differences were statistically significant ( t=3.457, 5.342 and 2.674, all P<0.05). After treatment the BMI and hemoglobin levels of CD patients were higher than those before treatment ((19.5±2.1) kg/m 2 vs. (18.3±1.8) kg/m 2, (117.2±10.3) g/L vs. (95.3±8.4) g/L), and the CRP level, CDAI score and SES-CD score were lower than those before treatment ((16.3±2.3) mg/L vs. (47.2±9.3) mg/L, 113.2±12.5 vs. 245.2±23.5, 5.0±2.1 vs. 10.0±4.3), and the differences were statistically significant ( t=2.090, 2.339, 2.432, 6.345 and 5.234, all P<0.05). The expression levels of TNF-α, IFN-γ, IL-2, IL-6, IL-8, IL-17A and their mRNA levels of healthy control group were lower than those of CD patients before treatment ((1.1±0.4) ng/L vs.(158.2±38.3) ng/L, (3.2±0.8) ng/L vs. (28.3±13.4) ng/L, (2.7±1.3) ng/L vs. (3.3±2.4) ng/L, (5.2±0.3) ng/L vs. (16.3±7.4) ng/L, (16.3±6.3) ng/L vs. (18.9±10.2) ng/L, (10.5±2.3) ng/L vs. (38.5±11.2) ng/L; 1.00±0.00 vs. 4.68±0.34, 7.83±0.32, 1.25±0.46, 8.36±0.44, 2.01±0.89 and 6.83±0.53, respectively), and the differences were statistically significant ( t=2.345, 6.456, 3.008, 4.009, 7.045, 10.223, 8.345, 11.235, 1.114, 12.334, 5.304 and 5.678, all P<0.05). After treatment the TNF-α, IFN-γ, IL-2, IL-6, IL-8, IL-17A expression levels and their mRNA levels of CD patients were lower than those before treatment ((106.4±29.9) ng/L vs. (158.2±38.3) ng/L, (25.7±10.8) ng/L vs. (28.3±13.4) ng/L, (2.9±1.7) ng/L vs. (3.3±2.4) ng/L, (15.4±4.2) ng/L vs. (16.3±7.4) ng/L, (17.2±8.7) ng/L vs. (18.9±10.2) ng/L, (29.9±12.7) ng/L vs. (38.5±11.2) ng/L, 2.45±0.21 vs. 4.68±0.34, 3.75±0.18 vs. 7.83±0.32, 1.09±0.22 vs. 1.25±0.46, 3.78±0.21 vs. 8.36±0.44, 1.67±0.33 vs. 2.01±0.89, 2.96±0.11 vs. 6.83±0.53), and the differences were statistically significant ( t=9.345, 2.456, 2.334, 2.090, 3.009, 8.345, 4.567, 6.445, 2.046, 7.774, 3.008 and 8.867, all P<0.05). The results of Western blotting showed that the expression levels of occudin, claudin-1, ZO-1 and JAM-A in the intestinal mucosa of CD patients before treatment were lower than those of the healthy control group (0.21±0.03 vs. 1.00±0.02, 0.17±0.07 vs. 1.00±0.01, 0.16±0.06 vs. 1.00±0.04, 0.26±0.08 vs. 1.03±0.04). After treatment the expression levels of occudin, claudin-1, ZO-1 and JAM- A mRNA in the intestinal mucosa of CD patients were higher than those before treatment (0.77±0.08 vs. 0.21±0.03, 0.69±0.08 vs. 0.17±0.07, 0.78±0.09 vs. 0.16 ±0.06, 0.72±0.07 vs. 0.26±0.08), and the differences were statistically significant ( t=4.567, 6.346, 5.557, 8.456, 9.678, 8.671, 10.456 and 7.456, all P<0.05). Conclusions:IFX can effectively relieve the disease activity and improve the nutritional status of CD patients. IFX maintains the expression of intestinal epithelial tight junction protein by reducing inflammatory response, and repairs the intestinal mucosal barrier of CD patients.

Objective:To detect the known hotspot mutations of GNAS in children with McCune-Albrigtht syndrome(MAS) by droplet digital PCR, and to explore its application value in the diagnosis of MAS.Methods:A total of 122 children with MAS were enrolled in the pediatric department of Ruijin Hospital Affiliated to Medical College of Shanghai Jiaotong University. For the known mutation hotspot of GNAS gene (R201H/C), dd-PCR, real-time fluorescent pyrophosphatic activation polymerase reaction (PAP) and second-generation sequencing were used to detect the presence of gene mutation and to analyse the relevance with the clinical features.Results:GNAS gene mutation was detected in 89 out of 122 children with MAS and 57 cases were found to have mutations. The positive rates of ddPCR, PAP, and second generation sequencing were 77.42%, 29.03%, and 56.25%, respectively. The GNAS gene mutation was detected in all classical triad patients. Among them, the positive rates of ddPCR in peripheral blood of typical and atypical children were 100% and 73.1% respectively, which were significantly higher than those of the other two methods. The detection rate of GNAS mutation in precocious puberty with bone lesions was higher than that in precocious puberty with skin lesions, suggesting that fibrous dysplasia with precocious puberty is an important basis for clinical diagnosis of MAS in children.Conclusion:Precocious puberty is the most common endocrine manifestation of MAS in children. Bone fibrous dysplasia with precocious puberty is an important factor in clinical diagnosis. ddPCR has high sensitivity, which can be helpful for molecular diagnosis of MAS.