ObjectiveTo evaluate the intraoperative neuromonitoring of the external branch of the superior laryngeal nerve (EBSLN) during Micooli's endoscopic thyroidectomy in order to avoid nerve injury.MethodsIn this study,36 patients with 56 nerves at risk were enrolled from February 2011 to September 2011.A positive signal is determined by observing contractions of the cricothyroid muscle to locate the EBSLN.The relationship between EBSLN and the upper pole of the thyroid or the inferior constrictor muscle was studied.The VHI-10 table was used for evaluation pre- and postoperatively. ResultsAll 56 nerves were located successfully,26 nerves(46.4％ ) crossed the superior thyroid artery more than 1 cm apart from the upper pole of the thyroid gland,while the other 30 nerves(53.6％ ) did less than 1 cm.In cases where the diameter was longer than 5 cm,the nerves crossed the artery at less than 1.0 cm from the upper pole in 73％ cases(P =0.006).There was no significant difference between VHI-10 results before and after surgery (P ＞ 0.05). ConclusionsIntraoperative neuromonitoring is useful and helpful in avoiding nerve injury by locating EBSLN.

Objective To evaluate the clinical value of breast localization needle placed via mammary ductoscopy in surgical treatment of patients with intraductal neoplasms. Methods In this study,76 patients with intraductal neoplasms diagnosized by mammary ductoscopy from January 2009 to March 2010 in the Second Affiliated Hospital of Soochow University were randomly divided into two groups.For methylene blue group,ducts with the lesion were marked by methylene blue injection before operation.For localization needle group,ducts were marked by localization needle placed via ductoscopy.The operative time,specimen weight,incision length and diagnostic coincidence rate were compared among the two groups. Results Compared to the methylene blue group,the localization needle group had a significantly shorter operative time (31 ± 8 min vs.42 ± 9 min),lighter specimen weight (1.51 ± 1.36 g vs.2.95 ± 2.07 g),and shorter incision (23.2 ± 7.8 mm vs.34.4 ± 7.1 mm).All the breast cancer cases dianosised by mammary ductoscopy were confirmed by postoperative pathology,but the localization needle group had a higher diagnostic coincidence rate than the methylene blue group (94.7% vs. 76.3%). Conclusion Localization needle under mammary ductoscopy is a reliable technique for localizing intraductal neolasms.The surgical excision guided by localization needle is accurate and less traumatic,and should be a routine method marking the tumor involved duct before operation.

Adolescent ; Adult ; Endoscopy ; methods ; Female ; Humans ; Male ; Middle Aged ; Nasal Septum ; abnormalities ; surgery ; Young Adult

Objective To evaluate the mechanism of external branch of the superior laryngeal nerve (EBSLN) injury during thyroid surgery as showed by intraoperative neuromonitoring.Methods 70 patients with 109 nerves at risk were enrolled in this study from March 2011 to October 2011.A positive signal is determined by observing contractions of the cricothyroid muscle.The relationship between EBSLN and the upper pole of the thyroid or the inferior constrictor muscle was studied.Results 108 nerves (99.1％) were located successfully,42 of which were visualized (38.9％).55 nerves (50.9％) crossed the superior thyroid artery more than 1 cm apart,while the other 53 nerves (49.1％) went less than 1 cm including 24 nerves(more than 0.5 cm,less than 1 cm) and 29 nerves (less than 0.5 cm) or coursed below the superior pole of the thyroid.The rate of the latter type was significantly elevated in patients with the top to botton diameter of the gland more than 5 cm.One patient suffered from impairing in the production of high tones postoperatively.Conclusions Intraoperative neuromonitoring is useful and helpful in providing instructive information for operations by locating EBSLN.

[Objective]To compare the clinical outcomes of fresh embryo transfer of the in vitro fertilization/intracytoplasmic sperm injection and embryo transfer(IVF/ICSI-ET)in different age groups as well as in different responders using gonadotropin-re-leasing hormone agonist(GnRH-a)long protocol or GnRH antagonist(GnRH-ant)protocol.[Methods]A retrospective analysis was performed on 737 IVF/ICSI cycles,including 386 cycles of GnRH-a long protocol(group A)and 351 cycles of GnRH-ant protocol (group B),from August 28,2015 to December 31,2016. Then all the cycles were divided into sub-groups by ages and retrieved oo-cyte numbers:group a1(<38 years),group a2(≥38 years);group b1(n≤5),group b2(6≤n≤15),group b3(n>15). The basic information of patients and clinical outcomes were compared.[Results](1)Comparable results were obtained from group A and group B in these following variables such as fertilization rate,normal fertilization rate,biochemical pregnancy rate and miscarriage rage. But the stimulation period,the total gonadotropin(Gn)dosage,estradiol(E2)level and endometrial thickness on the day of human chorionic gonadotropin(hCG)administration,number of oocytes retrieved and mature oocytes,ovarian hyperstimulation syn-drome(OHSS)rate,implantation rate and clinical pregnancy rate were significantly higher in group A than group B(P<0.05),and significantly higher cancellation rate of fresh embryo transfer was observed in group B(P<0.001).(2)When divided by ages,no mat-ter in sub-group a1 or sub-group a2,the implantation rate was slightly lower in GnRH-ant protocol than in GnRH-a long protocol, although they failed to reach significant difference(sub-group a1:32.6%vs 39.8%,P=0.067;sub-group a2:9.7%vs 17.9%,P=0.066). The clinical pregnancy rate was comparable using these two protocols in sub-group a1(54.8%vs 50.4%,P=0.429),but it was significantly lower by using GnRH-ant protocol than GnRH-a long protocol in sub-group a2(19.6%vs 39.1%,P=0.021).(3) When divided by numbers of oocytes retrieved,the implantation rate was significantly lower when using GnRH-ant protocol in sub-group b1(13.1%vs 26.0%,P=0.026),but we failed to observe significant differences in other two sub-groups. The clinical preg-nancy rates were comparable in all sub-groups ,whereas differed considerably in sub-group b1 (36.6% vs 19.3%,P = 0.056).[Conclusion]Overall,the implantation rate and clinical pregnancy rate were higher in GnRH-a long protocol than those in GnRH-ant protocol. Nevertheless,GnRH-ant protocol could reduce the dosage of Gn,shorten the treatment duration,and effectively reduce the occurrence of OHSS. There were similar pregnancy outcomes in two protocols for normal responders and high responders ,while for advanced patients or other poor responders,the implantation rate and clinical pregnancy rate were higher in GnRH-a protocol.

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are very two important pathogens that have coursed huge economic losses in swine production in worldwide. In this study,a vector pCMV-TJM containing the full-length cDNA clone of PRRSV attenuated strain TJM-F92 was firstly constructed by PCR method. Then a gene sequence containing Afl II/Mlu I e restriction enzyme sites and a transcription regulatory sequence for ORF6 (TRS6) was inserted be- tween ORF7 and 3'UTR, yielding a expression vector pCMV-TJM-TRS. Subsequently, a plasmid pCMV-TJM-Cap was constructed by cloning of PCV2 ORF2 gene into the unique sites Afl II /Mlu I of pCMV- TJM-TRS plasmid DNA. Then three recombinant PRRSV, rTJM, rTJM/TRS and rTJM/Cap, were rescued by transfection of pCMV-TJM, pCMV-TJM-TRS and pCMV-TJM-Cap into Marc-145 cells, respectively,and confirmed by the genome sequence, restriction enzyme digestion, Western Blot and IFA. They all had the molecular markers which was different from the parent virus. The growth characteristics of the rescued viruses were similar to that of parent virus. rTJM/Cap could also express efficiently PCV2 Cap protein in Marc-145 cells. At passage 8, it still had PCV2 ORF2 gene which examined by RT-PCR. It indicated that the full-length cDNA clone of PRRSV attenuated strain TJM-F92 and recombinant PRRSV rTJM/Cap expressing PCV2 Cap protein were successfully constructed. It made an important foundation for studying on the pathogenic mechanisms of PRRSV and PRRSV-PCV2 vaccine in the future.
Animals ; Capsid Proteins ; genetics ; immunology ; Cell Line ; Circoviridae Infections ; veterinary ; virology ; Circovirus ; classification ; genetics ; metabolism ; Gene Expression ; Open Reading Frames ; Porcine Reproductive and Respiratory Syndrome ; virology ; Porcine respiratory and reproductive syndrome virus ; genetics ; metabolism ; Recombination, Genetic ; Swine ; Swine Diseases ; virology ; Viral Vaccines ; genetics ; immunology

Objective To investigate the feasibility of Ad-hTGF-β1 transfected bone marrow mesenchymal stem cell(BMSCs) into chondrocytes differentiation and the change of TAZ mRNA. Methods Rats BMSCs were obtained and cultured by whole bone marrow method, and then the third-generation cells were seeded into cell culture plate, and divided into three groups:Ad-hTGF-β1 transfected group,Ad-EGFP transfected group and the control group. The control group was added in common medium without any treatment while the other two groups were respectively added in serum-free medium containing Ad-hTGF-β1 or that containing Ad-EGFP. Seven days later, real-time fluorescent quantitation PCR and Western blot were employed for detecting the expression of TGF-β1 ,while immunohistochemical and Western blot for the expression of collagen Ⅱ , and real-time fluorescent quantitation PCR for the expression of TAZ mRNA. Results Seven days after the transfection, real-time fluorescent quantitation PCR revealed that the average relative expression of TGF-β1 was:Ad-hTGF-β1 group 0.863, Ad-EGFP group 0.183, and the control group 0.180; The average relative expression of TAZ was:Ad-hTGF-β1 group 0.810, Ad-EGFP group 0.416, and the control group 0.366.The expression difference of TGF-β1 and TAZ were statistically significant (P ＜ 0.05). Western blot and immunohistochemical proved strong collagen Ⅱ expression in Ad-hTGF-β1 group while it was detected a little in the other two groups. Conclusion BMSCs could be successfully and stably induced into chondrocytes differentiation by Ad-hTGF-β1. Meanwhile, the mRNA of TAZ is up regulate during the differentiation,so it is suppose that TGF-β1 improve BMSCs into chondrocytes differentiation by TAZ.

ObjectiveTo rapidly detect the mutation frequency of DBC2 gene 7776C ＞ T in breast cancer and breast fibroadenoma by applying the mutation-sensitive molecular switch ( comprised of high-fidelity DNA polymerase and phosphorothioate-modified allele-specific primers) and agarose gel electrophoresis.Methods Allelic specific primers targeting mutation type and wild type were designed with the primers'3'terminal phosphorothioate modification.When the primers matched with the tissue DNA,the primers could be extended with highfidelity polymerase; when they mismatched with the tissue DNA,the primers could not be extended.DNA samples from 85 cases of breast cancer and 10 cases of breast fibroadenoma tissues were chosen and analyzed by PCR amplifications mediated by high-fidelity DNA polymerase.Gel imaging system was employed to make analysis of PCT products.ResultsThe mutation-sensitive molecular switch system showed that the mutation rate of 7776C ＞ T was 2.4％ ( 2/85 ) in the 85 cases of breast cancer,and no mutation was found in the 10 cases of breast fibroadenoma.ConclusionsThe mutation-sensitive molecular switch combined with agarose gel electrophoresis can rapidly detect the mutation of breast cancer DBC2 gene 7776C ＞ T.It is applicable in single nucleotide polymorphisms assay and has enormous application value in detecting gene mutation.

Objective To study the significance of the sample S/Co ratio when using a domestic reagent for anti-hepatitis C virus(HCV)antibody detection and to explore the procedure and standard of anti-HCV antibody diagnosis by using this domestic reagent.Methods Anti-HCV antibody was detected in 295 000 blood donors by a domestic anti-HCV reagent with enzyme-linked immunosorbent assay(ELISA)method and the reactive samples were tested again by ortho anti-HCV antibody reagent.The samples which anti-HCV antibodies were determined as positive by ortho anti-HCV rea- gent were examined by recombinant immunoblot assay(RIBA)reagent and 106 samples of them were also tested for HCV RNA.Results Six hundred and eighty-one samples were reactive in 295 000 samples screened by the domestic ELISA reagent,the reactive ratio was 0.23 %.Among the reactive samples screened by the domestic ELISA reagent,367 samples were determined as positive by ortho anti-HCV reagent while 66.2% of them showed a S/Co ratio≥3.8.The consistency rate between positive results determined by the domestic reagent and RIBA reagent respectively was 53.8%.For the samples showing S/Co ratio≥3.8 by ortho anti-HCV reagent,94.2% had a S/Co ratio≥8.0 when using the domestic ELISA reagent,while the percentage of samples showing S/Co ratio

Bone Marrow Cells ; metabolism ; pathology ; CD28 Antigens ; blood ; metabolism ; CD4 Antigens ; blood ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; Dendritic Cells, Follicular ; metabolism ; pathology ; Humans ; Immunoblastic Lymphadenopathy ; metabolism ; pathology ; Immunophenotyping ; methods ; Lymphoma, T-Cell ; metabolism ; pathology ; Neprilysin ; blood ; metabolism ; Receptors, Complement 3d ; blood ; metabolism ; fas Receptor ; blood ; metabolism