AIM:To investigate the function of macrophage differentiation antigen-1 (Mac-1) in receptor acti-vator of nuclear factor-κB ligand ( RANKL )-induced osteoclast differentiation and the mechanisms .METHODS: The spleen cells were isolated from 4-week-old C57BL/6J mice, and cultured with RANKL, macrophage colony-stimulating fac-tor and CD11b and CD18 antibodies for 1 week.The actin bundles were stained with rhodamine-labeled phalloidin, and nu-clei were stained with DAPI.CD11b and CD18 antibodies, lentivirus with interfering vector plasmid of target gene Itgam (encoding CD11b) and empty virus (control virus) were used to treat osteoclasts for 1 week, and then immunofluorescence staining was performed.CD11b antibody, lentivirus and control virus were used to treat osteoclasts for 1 week, and total protein was taken for Western blot .RESULTS:Lower multinuclear positive rates in the groups treated with CD 11b anti-body were observed than that in the groups treated with CD 18 antibody and control group .Lower immunofluorescence inten-sity of Syk, CD11b and NFATc1 was found in CD11b antibody group than that in control group .Lower Syk, CD11b and NFATc1 immunofluorescence intensity was also observed in lentivirus group than that in control virus group .The results of Western blot analysis showed that the protein levels of CD 11b, Syk, NFATc1, c-Fos and p-ERK/ERK in CD11b antibody group were decreased as compared control group .Compared with control virus group , the protein levels of CD11b, Syk, NFATc1, c-Fos and p-ERK/ERK in lentivirus group were also decreased .CONCLUSION:CD11b subunit of Mac-1 pro-motes osteoclast differentiation by up-regulating c-Fos, ERK activity and NFATc1.

Objective To construct the prokaryotic expression vector bearing fusion gene NT4-ADNF-9 and lay foundation for further study on genetic therapy of neuraseusory deafness. Methods By means of asymmetrical prince/ template, double stranded eDNA of activity dependent neurotrophic factor-9 (ADNF-9) was obtained, which included restriction enzymes sites on the two extremities. ADNF-9 eDNA was ligated to the signal and leader peptides of nenrotrophin 4 (NT4), and the fusion gene was named NT4-ADNF-9. Then it was suheluned into prokaryotic expression vector pBV220, and called pBV220/ NT4-ADNF-9. Results Evidences of DNA sequence analysis and restrtction enzymes digestion showed that we recombined ADNF-9 eDNA to the 3'terminal of the signal and leader peptides of NT4, and the fusion gene was subcluned into pBV220 successfully. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of dorsal root ganglia (DRG) of embryonic day-8 cbicken neurons as compared to the control. Conclusion Prokaryotic expression vector pBV220/NT4-ADNF-9 can be constructed successfully and the bioactivtty is satisfactory.

Objective To design the ISO15189 management information middleware based on laboratory information management system (LIS) and evaluate its application.Methods The ISO15189 management information middleware was designed based on LIS and the middleware technology as underlaying platform,and the multifunction management of LIS was implemented by the data warehouse technology and the data cleaning and extraction technology.Results The designed middleware made LIS implement the functions such as data exchange and transmission,data sharing,and data intelligence processing ISO15189 management information across multiple systems.It constructed a data warehouse by extracting,transforming,and loading useful data from multiple profession systems in laboratories,and flexibly presented data to the users by operating multidimensional data set from various aspects.Conclusion The ISO15189 management information middleware based on LIS has high value in the management of ISO15189 accredit,which may make the management work more standard,efficiency,and intelligence.

Objective To discuss the clinical value of new- utility anal vacuum tube in preventing anastomotic fistula after colorectal cancer anterior resection. Methods Ninety-six patients having underwent the colorectal cancer anterior resection were divided into new-utility anal vacuum tube group and normal anal vacuum tube group by random digits table method with 48 cases each. The complication, anus exhaust time and postoperative drainage volume were compared between 2 groups. Results The incidences of tube defluxion, proctalgia, anus skin damage, bed sheet pollution and anastomotic fistula in new-utility anal vacuum tube group were significantly lower than those in normal anal vacuum tube group: 6.25% (3/48) vs. 31.25% (15/48), 10.42% (5/48) vs. 41.67% (20/48), 0 vs. 25.00%(12/48), 6.25%(3/48) vs. 60.42%(29/48), 2.08%(1/48) vs. 12.50%(6/48), the anus exhaust time was significantly shorter than that in normal anal vacuum tube group:(44.1 ± 8.9) h vs. (48.9 ± 9.6) h, the postoperative drainage volumes form the first day to fifth day were significantly more than those in normal anal vacuum tube group: (31.2 ± 15.1) ml vs. (15.6 ± 8.2) ml, (25.3 ± 13.2) ml vs. (15.8 ± 6.5) ml, (15.6 ± 9.1) ml vs. (10.3 ± 4.5) ml, (104.3 ± 38.2) ml vs. (90.6 ± 12.3) ml and (93.7 ± 32.5) ml vs. (80.7 ± 18.9) ml, and there were statistical differences (P<0.01 or <0.05). The patients in new-utility anal vacuum tube group had different symptoms, but patients could tolerate. Conclusions The new-utility anal vacuum tube can reduce the incidence of anastomotic fistula, and be safe and reliable, which is worthy of wide application.

BACKGROUND: Further studies are needed to understand the cytobiological character, functional regulation, gene changes and expression difference of CD34~+ and CD133~+ stem cells induced by basic fibroblast growth factor (bFGF) using gene chip. OBJECTIVE: To compare the differences of gene expression and the response to bFGF of human umbilical cord CD34~+ and CD133~+ cells, and to explore gene expression changes of bFGF-induced umbilical cord CD34~+ and CD133~+ hematopotic stem cells/hemapoietic progenitor cells in vitro. METHODS: Human umbilical cord blood CD34~+ and CD133~+ cells were isolated and purified by MiniMACS immunomagnetic beads selection. The CD34~+ and CD133~+cells were cultured for 10 to 15 days in DMEM/F12 medium, supplemented with bFGF and B27. Total RNA from these cells was extracted and the genetic level of these cells was performed using Oligo GEArray(r) chip and GEArray software. Selected rate of CD34~+ and CD133~+ hematopoietic stem cells was detected using flow cytometry. CD34~+ and CD133~+ cell morphological changes were measured before and after bFGF induction. The concentration and purity of RNA were determined by agarose gel electrophoresis degeneration. Gene-chip test results were analyzed.

Objective To investigate the protective function of edaravone in the compressed spinal cord.Methods There were 150 rabbits enrolled in each group in the experiment.Rabbits in both operation group and edaravone (EDA) treating group received mild spinal cord compressionby setting a flap head screw between C6 C7 after the neck.The spinal cord decompression was conducted seven days later.After 6 hours,rabbits in the EDA treating group were injected with a large amount of EDA through ear border veins,while the rabbits in the operation group only received 0.9％ sodium chloride injection.The transmission electron microscope was used to observe the apoptotic bodies at 1 day,3 days and 7 days after compression,and 1 day,3 days,7 days,and 14 days after decompression.Flow cytometry was used to test the rate of apoptosis of spinal cord cells.Immunohistochemistry was used to test the expression of Bax protein that is related to apoptosis.Results The neuronal apoptosis appeared after compression in both operation group and EDA-treating group.The Basso Beattie Bresnahan (BBB) score,neuronal apoptosis rates,and Bax protein expressions in both groups were statistically different (P ＜ 0.05) when the spinal cord was compressed in the first day and the third day,while there was no statistically different when spinal cord compressed at the seventh day (P ＞ 0.05).After decompression of the spinal cord,the BBB score,neuronal apoptosis rates,and Bax protein expressions in both groups were becoming lower at the seventh day (P ＜0.05).Conclusions EDA has protective function for compressed spinal cord.However,only the compression of spinal cord compression period of sufficient decompression can fundamentally protect the spinal cord.

OBJECTIVE: To investigate the effect of IL-17 and IL-23 in the pathogenesis of allergic rhinitis(AR) and non. allergic rhinitis(NAR). METHOD: Selected 156 cases of patients with allergic rhinitis (AR group) and 59 cases of patients with non-allergic rhinitis (NAR group), 60 cases of healthy people (control group). All cases in AR group and NAR groups were evaluated by a visual analog scale (VAS) score of nasal symptoms. Collected peripheral blood and nasal secretions in all cases and then detected IL-17 and IL-23 expression levels. RESULT: There was no significant difference in VAS score of AR group and NAR group (P>0. 05). IL-17 and IL-23 levels of serum and nasal secretions in AR group and NAR group were both higher than control group, with a highly significant difference (P <0. 05). The research showed a clear correlation between expression of IL-17 and IL-23 both in serum and nasal secretions of AR group and NAR(P<0. 05). CONCLUSION IL-17 and IL-23 may be important cytokines and IL-23/IL-17 pathway may play a significant role in the pathogenesis of allergic rhinitis and non-allergic rhinitis.
Case-Control Studies ; Humans ; Interleukin-17 ; blood ; metabolism ; Interleukin-23 ; blood ; metabolism ; Nasal Mucosa ; metabolism ; Rhinitis ; blood ; metabolism ; Rhinitis, Allergic ; blood ; metabolism

Objective To investigate the impact of transvaginal natural orifice transluminal endo-scopic surgery(NOTES)-assisted laparoscopic nephrectomy on female sexual function and quality of life . Methods This was a prospective study on the change of female sexual function and quality of life of female patients who underwent transvaginal NOTES-assisted laparoscopic nephrectomy from May .2011 to Nov. 2012.A total of 42 cases were included in this study (28 of them with severe hydronephrosis , non-functio-ning kidney , 11 with pyelonephrosis , 1 with renal tuberculosis , 1 with duplex kidney complicated with hy-dronephrosis, and 1 with renal angiomyolipoma ).The mean age was 36.9±5.3 (26-45) years, and the mean body mass index was 21.7±2.6 (14.7 to 27.1) kg/m2.Twenty-four cases were operated on the left side, 18 cases on the right .The female sexual function and quality of life were assessed before and 4 months, 7 months and 1 year after surgery using the Female Sexual Function Index (FSFI) questionnaire and the MOS 36-item Short-Form Health Survey (SF-36), respectively. Results The mean FSFI of 42 cases preoperatively and 4 months, 7 months and 1 yr postoperatively were 27.74 ±4.34, 27.19 ±4.49, 28.54±4.23, and 28.68 ±4.19, respectively.There was no statistically significant difference among them (F=1.111, P=0.346).Compared with that of preoperation , the physical function, vitality, metal health, body pain, and general health of the patients were improved , but the role-physical, role-emotion and social function were not improved at postoperative month 4 and month 7 (P<0.05).Each item of SF-36 was im-proved after postoperative 1 year ( P<0.05) . Conclusions Transvaginal NOTES-assisted laparoscopic ne-phrectomy does not cause negative effect on the female sexual function .The quality of life can be improved after operation .The physical function is improved at early stage , and the psychological function as well .

OBJECTIVE: To construct an universal recombinate adeno-associated virus (AAV), rAAV-NT4-ADNF-9, and to detect the ability of transfection of the rAAV vector into cochlea in vitro. METHOD: pSSVHG-CMV-ADNF-9 plasmid was introduced into 293 cell by method of Ca3 (PO4)2 using three plasmids of pSSHG-CMV-NT4-ADNF-9, pFG140 and pAAV/Ad. The recombinate adeno-associated virus (rAAV) was harvested, and the titrations of the rAAV concentrated was detected by dot-blot test. Isolate and culture the cochlear hair cell of SD rats newly born in vitro. The rAAV-NT4-ADNF-9 was added to the medium while plating. Cochlear were collected 24 h after cultivation for RT-PCR to detect the transfection of rAAV-NT4-ADNF-9. RESULT: The titration of rAAV stock produced 2 x 10(16) total particles/L, which showed that rAAV-NT4-ADNF-9 was constructed successfully. The cochlear hair cell of SD rats newly born was isolated and cultured in vitro successfully. It certified that rAAV-NT4-ADNF-9 was able to transfect into cochlear and express secretory NT4-ADNF-9 peptide by RT-PCR. CONCLUSION The rAAV vector constructed in this paper, rAAV-NT4-ADNF-9, can transfer into cochlear cultured in vitro, which layed a foundation of further research for gene therapy.
Animals ; Cells, Cultured ; Cochlea ; cytology ; Dependovirus ; genetics ; Genetic Vectors ; Hair Cells, Auditory ; cytology ; Nerve Tissue Proteins ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

Objective To describe the initial clinical experience of transvaginal NOTES-assisted laparoscopic nephrectomy. Methods From May to June 2010, 4 female patients with non-functioning kidney and 1 female patient with renal tuberculosis underwent transvaginal NOTES-assisted laparoscopic nephrectomy. The median age was 41 years (range 36 to 63). Three lesions were on the right side, and 2 on the left. After induction of general anesthesia, the patients were positioned in lithotomy with ipsilateral lumbar at 60° angle to the operating table. A 5 mm Trocar and a 10 mm Trocar were placed through the umbilicus, and a 10 mm Trocar for 30° laparoscope was placed through the posterior vaginal fornix under direct vision. The kidneys were put into a home-made bag and removed via the incision of posterior vaginal fornix after there were completely resected. Results The 5 procedures were successfully finished. The median operative time was 190 (range 150 to 260)min. The median estimated blood loss was 185 (range 150 to 210) ml. There were no intra-operative or postoperative complications. The patients resumed ambulation 1 day after surgery, and resumed nutrition 2 days after surgery. The drainages were removed on postoperactive day 3. The patients were discharged on postoperative day 7. The incision in vagina healed well. Conclusions Transvaginal NOTES-assisted laparoscopic nephrectomy is feasible and reproducible. This novel technique may provide additional benefits related to postoperative pain, recovery, and cosmetic results even when compared to today's minimally invasive procedures.