Brown-Sequard Syndrome ; etiology ; Cervical Vertebrae ; Humans ; Intervertebral Disc Displacement ; complications ; surgery ; Male ; Middle Aged

OBJECTIVETo search for the optimal strategies for sperm collection from the patient with temporary penile erectile dysfunction (ED) on the day of oocyte pick-up ( OPU) in in vitro fertilization embryo transfer (IVF-ET).

METHODSWe retrospectively analyzed 93 cases of temporary ED on the OPU day of IVF-ET from January 2011 to May 2014, with fresh semen for 45 cases (group A), cryopreserved sperm before oocyte retrieval for 30 cases (group B), and frozen oocytes for 18 cases (group C). Group A was again subdivided into A1 (n = 18) and A2 n = 27) , the former intervened with oral sildenafil while the latter left untreated. We compared the rates of fertilization, high-quality embryo, and pregnancy among different groups.

RESULTSNo statistically significant differences were found among groups A, B and C in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and rates of cleavage, or in the percentages of normal fertilization (80.78% vs 80.43% vs 84.77%), high-quality embryo (53.27% vs 52.97% vs 47.69%) and pregnancy (60.00% vs 56.77% vs 44.44%) (all P > 0.05). The rate of 3PN was markedly lower in group C (0.63%) than in A (9. 61%) and B (4.34%) (P < 0.05). There were no significant differences between groups A1 and A2 in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and the rates of fertilization, cleavage, high-quality embryo, and pregnancy (all P > 0.05).

CONCLUSIONOn the OPU day of IVF-ET, oral sildenafil can help temporary ED men to achieve penile erection and ejaculation without affecting the outcomes of assisted reproduction. Cryopreserved sperm can be used in case of predicted temporary ED and frozen oocytes can also be employed if sperm retrieval fails. However, to avoid puncture injury to the epididymis or testis, fresh semen should be the first choice.

Cryopreservation ; Embryo Transfer ; Erectile Dysfunction ; drug therapy ; physiopathology ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Male ; Oocyte Retrieval ; Oocytes ; Penis ; Pregnancy ; Retrospective Studies ; Sildenafil Citrate ; therapeutic use ; Sperm Retrieval ; Spermatozoa

Female ; Humans ; Kidney Neoplasms ; complications ; pathology ; Ovarian Neoplasms ; pathology ; Wilms Tumor ; complications ; pathology ; Young Adult

Objective To establish a mouse model of circulating tumor cells (CTCs) by applying mouse hepatoma Hapa 1-6 cells.Methods 108 healthy male C57BL/6 mice were randomly divided into 3 groups according to their body weights.Hepa 1-6 cell suspension was intravenously injected to each mouse in the three groups at a concentration of 1×106,5×106 and 1×107/mL,0.2 mL per mouse,respectively.Blood samples were collected from the mice on the 1st,5th,9th,13th,17th and 21st days after tumor cell injection.The number,ratio and relative inhibition rate of CTCs were calculated in 20,000 nucleated cells.The mortality of mice was recorded.②80 male C57BL/6 mice were averaged into 2 groups according to their body weight: control and sorafenib tosylate groups.0.2 mL of Hepa 1-6 single cell suspension was injected to each mouse through the caudal vein at a concentration of 5×106/mL.The mice were gavaged with sorafenib tosylate (50 mg/kg) for 21 days and blood samples were collected at the 3rd,8th,15th,and 21st days for CTC assessment.Results For the 1×106/mL group,the CTC inhibition rate was 25.1%,18.1%,8.9%,4.4%,2.9% and 0.3% on the 1st,5th,9th,13th,17th and 21st days,respectively,and all the mice were alive.For the 5×106/mL group,the CTC inhibition rate was 40.4%,35.4%,15.4%,9.0%,6.6% and 4.1% on the 1st,5th,9th,13th,17th and 21st days,respectively,and all the mice were alive.For the 1×107/mL group,the CTC inhibition rate was 39.1% and 33.5% on the 1st and 5th days,respectively.Some mice died immediately after intravenous injection and all mice died within 7 days.②The relative clearance of CTCs was-7.5%,4.6%,55.3% and-94.5% on the 3rd,8th,15th and 21st days of sorafenib tosylate administration.Compared with the control group,there were significant differences among the three groups (P<0.05 or P<0.01).Conclusions A mouse model of circulating hepatoma cells has been established by intravenous injection of 0.2 mL of 5×106/mL mouse Hepa 1-6 cell suspension.This mouse model can be used for screening and evaluation of drugs for circulating tumor cell inhibition.

Objective To study the clinical data of patients with locally advanced cervical cancer(LACC)treated with preoperative radiotherapy plus chemotherapy combined with surgery. Methods Seventy patients with LACC(stage ⅠB2,ⅡA2 andⅡB)who were treated in our hospital from January 2012 to December 2016 were selected in this study.All the cases were randomly divided into two groups.The observation group(n=35)was treated with three-dimensional intracavitary brachytherapy plus chemotherapy combined with surgery, while the control group (n=35) was treated with radical radiotherapy.Patients of the two groups were followed up after the treatment.The recent and long term complications were recorded and observed in the two groups.Meanwhile,the survival curves were drawn by Kaplan-Meier,and the difference of total survival rate was compared with the Log-rank method between the two groups. Results The incidence of rectal reaction was less in the observation group than that in the control group(14.3% vs.37.1%,χ 2=4.786,P<0.05).There were no significant differences in bone marrow suppression,gastrointestinal reaction and bladder reaction between the two groups (P>0.05). The incidence rates of radioactive bladder injury (8.6% vs. 31.4%) and radionuclide injury (11.4% vs. 34.2%) were less in the observation group than those in the control group(χ2=5.714 and 5.185 respectively,P<0.05).The survival analysis showed that there was no significant difference in the total survival rate between the two groups of stageⅠB2+ⅡA2 and stageⅡB patients(Log rank χ2=0.081 and 0.376,P>0.05).Conclusion For patients with locally advanced cervical cancer, preoperative radiotherapy plus chemotherapy combined with surgery can reduce the incidence of related complications, meanwhile, there is no significant difference in the total survival rate and the mean survival time after the treatment of the two methods.

The aim of this study was to obtain the capsid protein VP2 of human bocavirus (HBoV) identified in Beijing recently and construct virus-like particles (VLPs) in insect cells for further study of this virus. The full-length VP2 gene of HBoV from BJ3722 was inserted into the baculovirus expression transfer vector (pFastBac 1) to obtain the recombinant Bacmid, and generation of recombinant baculoviruses was followed by transfection of the recombinant Bacmid into insect cells. Then the recombinant VP2 protein was recognized by SDS-PAGE using Coomassie-blue staining and Western blot using hyper-immune serum against VP2 of HBoV from rabbit. The recombinant baculoviruses were harvested and amplified to gain large amounts of viruses with high titers to infect insect cells at a multiplicity of infection (MOI) of 0. 5. After 7-10 days or 4-5 days of the infection, the supernatants of culture or the cell lysates treated with lysing solution were harvested, and ultracentrifuged twice through 40% sucrose cushion to obtain purified VLPs, which were followed by Western blot and IFA for VLPs' composition and specificity analysis, by electron microscopy for VLPs' morphologic structure. The recombinant VP2 protein with molecular weight of approximately 61 kD expressed in recombinant baculoviruses was recognized by SDS-PAGE using Coomassie-blue staining and Western blot. The presence of VP2 on VLPs was demonstrated by Western blot and IFA from samples collected during the purification of VLPs from the supernatants of culture or the cell lysates, and the expression of VP2 in insect cells led to the formation of VLPs which formed the typical icosahedral appearance of parvoviruses with a diameter of approximately 20 nm. In conclusion, the recombinant baculoviruses were constructed, the HBoV VP2 protein was expressed in insect cells with high specific antigenicity and VLPs was formed successfully.
Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Fluorescent Antibody Technique, Indirect ; Human bocavirus ; genetics ; metabolism ; Microscopy, Electron, Transmission ; Polymerase Chain Reaction ; Spodoptera ; Virion ; genetics ; metabolism ; ultrastructure

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.
Acute Disease ; Child, Preschool ; China ; Female ; Genome, Viral ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomavirus ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Viral Proteins ; genetics

Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.
Chromatography, Affinity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Antibodies ; biosynthesis ; genetics ; isolation & purification ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; genetics ; isolation & purification ; Pichia ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification