Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID₅₀, respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 μg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 μg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10⁴ and 1×10⁵. The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.

Objective To investigate the relationship of relative telomere length in cumulus cells(CCs) with oocytes at different mature stages and the outcome of intracytoplasmic sperm injection and embryo transfer (ICSI-ET). Methods Oocyte-cumulus complex samples were collected from 92 patients undergoing ICSI-ET and patients were divided into group A including 55 women≤35 years and group B 37 women>35 years. The embryonic development ability and the result of clinical pregnancy were recorded in different groups. DNA was extracted from CCs and assessed for telomere length by real-time quantitative PCR. Results In ICSI-ET,the relative telomere length of CCs gradually shortened with the age.There was a significantly longer telomere length of CCs in the preg-nant subgroup than that in the non-pregnant subgroup in the two groups(P < 0.05). Conclusion The telomere length of CCs may be associated with the pregnancy outcome in ICSI-ET.

Objective To investigate the differential diagnostic value of computed tomography (CT) combined with serum tumor markers in borderline ovarian tumors (BOT) and benign epithelial ovarian tumors (BET).Methods The CT data in 28 patients with BOT and 41 patients with BET,both confirmed by surgery and pathological,were analyzed retrospectively.Their preoperative serum carbohydrate antigen 125 (CA125),human epididymis secretory protein 4 (HE4) and carcinoembryonic antigen (CEA) detection results were collected.The CT images features and serum tumor markers levels were compared between the two groups.Results The difference in the appearance rate of tumor solid composition,thick septum and wall nodule between the two groups had statistical significance (x2 =25.135,5.240,5.066,P＜0.05).The serum CA125 level had statistical difference between the two groups (Z=3.202,P＜0.05),while serum HE4 and CEA levels had no statistically significant difference between the two groups(Z=0.330,1.122,P＞0.05).The optimal critical value,sensitivity and specificity of serum CA125 level in differential diagnosis of two kinds of tumor was 42.45 U/mL,53.6％ and 85.4％.The overall diagnostic rate of solid composition and thick septum for diagnosing the two kinds of tumor was 78.5 ％.The overall diagnostic rate of solid composition,thick septum and CA125 level for diagnosing the two kinds of tumor was 81.2％.Conclusion The appearance of solid composition,thick septum and serum CA125 level increase in epithelial ovarian tumor may help to identify BOT and BET.

Objective: To evaluate the clinical effect of pereutaneous vertbroplasty (PVP) combined with implantation of iodine-125 (125 I)radioactive particle in the treatment of vertebral metastasis,and to provide basis for the treatment of vertebral metastasis.Methods:A total of 69 patients with vertebral metastasis were divided into test group (n=32)and control group (n=37);the patients in test group were treated with PVP comined with implantation 125 I radioactive particle and the patients in control group were treated with PVP only.The heights of anterior and posterior vertebral bodies of the patients before and after treatment were detected by X-ray.The numerical rating scale (NRS)scores,pain relief rate and the incidence of surgical complications of the patients were recorded before operation and 1 d,1 week,1 month,3 months,and 6 months after operation.Results:The operation was successfully performed in all the patients without local bleeding;there were no movement dysfunction and nerve compression phenomenon.There was no leakage of bone cement.All the 125 I radioactive particles located well and there was no particle obscission.The heights of vertebral bodies of the patients in two groups after operation were increased compared with before operation (P <0.05).The NRS scores of the patients in two groups s at 1 d,1 week,1 month,3 months,6 months after operation were significantly decreased compared with before operation (P <0.05);compared with control group,the NRS scores of the patients in test group at 1 d,1 week, 1 month,3 months,6 months after operation were decreased (P <0.05).The incidence of pulmonary embolism or radiation myelitis complications was about 4.3% in 69 patients.Compared with control group,the difference in the incidence of complications of the patients in test group was not significant (P < 0.05 ).Conclusion:PVP combined with 125 I radioactive particle implantation is a safe and effective method in the treatment of vertebral metastasis,which can relieve the pain of the patients obviously compared with PVP.

OBJECTIVE To assess the association of single nucleotide polymorphisms (SNPs) of susceptibility genes of type 2 diabetes mellitus (T2DM) with liability to gout among ethnic Han Chinese males from coastal region of Shandong province. METHODS Seven SNPs within the susceptibility genes of T2DM, including rs10773971(G/C) and rs4766398(G/C) of WNT5B gene, rs10225163(G/C) of JAZF1 gene, rs2069590(T/A) of BDKRB2 gene, rs5745709(G/A) of HGF gene, rs1991914(C/A) of OTOP1 gene and rs2236479(G/A) of COL18A1 gene, were typed with a custom-made Illumina GoldenGate Genotyping assay in 480 male patients with gout and 480 male controls. Potential association was assessed with the chi-square test. RESULTS No significant difference was detected for the 7 selected SNPs in terms of genotypic and allelic frequencies (P > 0.05). When age and body mass index (BMI) were adjusted, the 7 genetic variants still showed no significant association with gout. CONCLUSION The genotypes of the 7 selected SNPs are not associated with gout in ethnic Han Chinese male patients from the coastal region of Shandong province. However, the results need to be replicated in larger sets of patients collected from other regions and populations.
Adult ; Aged ; China ; ethnology ; Diabetes Mellitus, Type 2 ; genetics ; Ethnic Groups ; Genetic Predisposition to Disease ; Gout ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Objective To investigate the diagnostic value of ultrasound and MRI in fetal bronchopulmonary sequestration (BPS). Methods The 7 pregnant women with suspected fetal BPS were examined with a 1.5 T MR unit within 24 h after prenatal ultrasound in Hubei Maternal and Children's Hospital during July 2013 to February 2015. The imaging protocol included half-fourier acquisition single shot turbo SE (HASTE), true fast imaging with steady state precession (True FISP) in axial, frontal and sagittal planes relative to the fetal thorax. Prenatal MRI findings have been compared with postnatal enhanced computed tomography or biopsy. Results The locations of BPS were in left side in 5 cases and in right side in 2 cases. One case was complicated with congenital cystic adenomatoid malformation (CCAM) of lung. Ultrasound showed the intrathoracic mass as a hyperechoic lesion and the feeding artery could be found by Doppler ultrasonography. T2WI could reveal not only the hyperintense lesions with clear boundary, but also the hypointense feeding artery originating from systemic circulation. Compared with pathological examination or enhanced CT, both of the ultrasound and the MRI could locate the lesions;however 2 feeding arteries were misjudged. Conclusions Prenatal ultrasound is the first-choice diagnostic modality for BPS. MRI can demonstrate the location, morphology and the feeding arteries of the fetal BPS, and also estimate the volume of normal lungs, which could be an important supplement to prenatal ultrasound in prenatal diagnosis and prognostic prediction of BPS.

Objective To discuss the value of prenatal ultrasound and MRI in diagnosis of fetal agenesis of corpus callosum (ACC). Methods Sixty-seven fetuses from Hubei Maternal and Children's Hospital performed fetal MRI from July 2013 to December 2014 were included in this study. All fetuses (67 cases) with suspected ACC were studied with a 1.5T MR unit within 3 days after ultrasound examination. Prenatal ultrasound and MRI findings were studied. Results All the 67 ACCs previously suspected on ultrasound were confirmed by MRI. Among the 67 ACCs, 58 cases were complete ACC and 9 cases were partial ACC. Corpus callosum body and/or splenium absence was found in all 9 partial ACC cases. In all cases, on MRI, corpus callosum complete or partial absence was showed on the median sagittal images, and mild to moderate ventriculomegaly and abnormal morphology in lateral ventricle was shown on the axial or coronary images. Conclusions MRI has high value in diagnosis of ACC. When ACC is found or suspected by ultrasonophy, MRI examination is suggested to confirm the diagnosis.

ObjectiveTo explore the imaging features of fetal tuberous sclerosis complex by ultrasonography and magnetic resonance imaging.MethodsRetrospective analysis on the imaging characteristics of the 10 cases of fetuses confirmed as tuberous sclerosis complex who were examined in Hubei Maternal and Child Healthcare Hospital in July 2013 to December 2014 by ultrasonography and MRI, which was compared with the pathological data of specimens and follow-up after birth.ResultsEighteen cases were diagnosed as fetal cardiac rhabdomyoma by ultrasonography among all of the 996 fetuses, in which lesions were located on the ventricular wall near septum or elsewhere in the heart cavity on ultrasonography. Fetal cardiac rhabdomyoma was characterized by circular, homogeneous high echo (singleton in 4 cases, multiple in 14 cases). Among them no subependymal nodule was found by ultrasonographic. Ten cases of subependymal nodule were found by magnetic resonance imaging, which were diagnosed as tuberous sclerosis complex with cardiac rhabdomyomas, including 3 cases of brain subcortical tubers. The subependymal nodules under the lateral ventricle wall showed characteristic low signal nodules on T2WI, protruding from the ependymal surface. Of 18 cases, only 4 cases of fetal cardiac rhabdomyoma were found by MRI. Nine cases of ifnally had termination of pregnancy. Two cases were conifrmed as cardiac rhabdomyoma with intracranial nodules after pathological examination, and 1 case was conifrmed as tuberous sclerosis complex after birth. ConclusionsPrenatal ultrasonography can diagnose fetal cardiac rhabdomyoma successfully, and MRI can diagnose the fetal brain nodules sensitively. Once ultrasonography finds cardiac rhabdomyoma, it may be promising to diagnose fetal tuberous sclerosis complex by ultrasonography combined with MRI.

BACKGROUND:Superparamagnetic iron oxide nanoparticles (Fe3O4 NPs) have been widely used in MRI. It is vital to prepare the superparamagnetic MRI contrast agent with high stability, biocompatibility and tumor targeting in order to prevent the aggregation of Fe 3 O 4 NPs and realize the high-precision diagnose of tumor. OBJECTIVE:To prepare the amphiphilic superparamagnetic composite particles with tumor targeting mediated by folate receptor. METHODS:The stable amphiphilic superparamagnetic composite particles with tumor targeting function were prepared by coating the Fe3O4 NPs with a Pluronic F127-folic acid conjugate, which was synthesized via an esterification reaction between the carboxyl group of the tumor targeting molecule, folic acid and the hydroxyl group of an amphiphilic triblock copolymer, Pluronic F127. The resultant Pluronic F127-folic acid-Fe3O4 composite particles were characterized by transmission electron microscopy, Fourier transform infrared-spectra, UV-vis absorption spectra, thermal gravimetric analysis, vibrating sample magnetometer and T2-weighted imaging. WST assay was used to characterize their cytotoxicity preliminarily. RESULTS AND CONCLUSION:The Pluronic F127-folic acid conjugates were prepared via esterification reaction. Then Fe 3 O 4 NPs were wrapped with Pluronic F127-folic acid to result in the superparamagnetic composite particles with wel dispersion and biocompatibility. The size of most superparamagnetic composite particles was less than 200 nm and the size of Fe 3 O 4 core was 10-20 nm from the observation of transmission electron microscopy. The results from the Fourier transform infrared-spectra and UV-vis absorption spectroscop confirmed that folic acid molecules were modified on the surface of the superparamagnetic composite particles successful y. The mass ratio of Pluronic F127-folic acid conjugate was determined by thermal gravimetric analysis as 27.2 wt%in the resultant Pluronic F127-folic acid-Fe 3 O 4 composite particles. The saturated magnetic intensity of the superparamagnetic composite particles was 47.35 emu/g by vibrating sample magnetometer and the relaxation rate was 0.025×106 mol/s from MRI. The WST assay showed the negligible cellcytotoxicity of Pluronic F127-folic acid-Fe3O4. The superparamagnetic composite particles have potential application as the MRI contrast agents with tumor targeting, and the Pluronic F127-folic acid-Fe 3 O 4 composite particles is expected to be used as a MRI contrast agent for tumor targeting.

OBJECTIVETo evaluate the long-term clinical outcomes of fractional flow reserve (FFR)-guided versus intravascular ultrasound (IVUS)-guided percutaneous coronary intervention (PCI) for intermediate coronary lesions.

METHODSA total of 226 patients with 293 intermediate coronary artery lesions (stenosis of 40%-70%) confirmed by coronary angiography were randomized into 3 groups to undergo PCI for a minimal lumen cross sectional area (MLA)<4 mm(2) (IVUS group, 98 lesions) or for a FFR<0.80 (FFR group, 101 lesions), or to receive standard medical treatment (medication group, 94 lesions). The primary outcome was major adverse cardiac events including death, myocardial infarction, and ischemia-driven target vessel revascularization at 1 year after the index procedure.

RESULTSThe baseline percent diameter stenosis and lesion length were similar between the 3 groups, but more patients in IVUS group than in FFR group received PCI (P<0.001). No significant difference was found in the incidence of major adverse cardiac events between the 3 groups (P=0.182).

CONCLUSIONBoth FFR- and IVUS-guided PCI strategy for intermediate coronary artery disease are associated with favorable outcomes, but IVUS-guided PCI based on the single index of MLA can increase the rate of revascularization therapy.