To explore a new way of constructing bioartificial renal tubule assist device (RAD) in vitro and its function of transporting sodium (Na(+)) and glucose and to evaluate the application of atomic force microscope in the RAD construction, rat renal tubular epithelial cell line NRK-52E was cultured in vitro, seeded onto the outer surfaces of hollow fibers in a bioreactor, and then cultured for two weeks to construct RAD. Bioreactor hollow fibers without NRK-52E cells were used as control. The morphologies of attached cells were observed with scanning electron microscope, and the junctions of cells and polysulfone membrane were observed with atomic force microscope. Transportation of Na(+) and glucose was measured. Oubaine and phlorizin were used to inhibit the transporting property. The results showed that NRK-52E cells and polysulfone membrane were closely linked, as observed under atomic force microscope. After exposure to oubaine and phlorizin, transporting rates of Na(+) and glucose were decreased significantly in the RAD group as compared with that in the control group (P<0.01). Furthermore, when the inhibitors were removed, transportation of Na(+) and glucose was restored. It is concluded that a new RAD was constructed successfully in vitro, and it is able to selectively transport Na(+) and glucose.

The effects of vasectomy on the reproductive organs in various species are controversial. This study investigated the morphological change and apoptosis of the testis, epididymis, and vas deferens in beagle dogs 12 months after vasectomy. The male beagles were divided into two groups: vasectomized and sham-operated groups (n=5 in each). Histopathological, ultrastructural, and TUNEL evaluation of the changes in the testis, epididymis, and ductus deferens of each animal were conducted 12 months after surgery. The mean lumen diameter, cellular thickness, mean interstitial distance, and lumen area fraction of each seminiferous tubule and ductus epididymis were measured by stereological analysis. The results showed that, compared with the sham-operated group, the seminiferous tubular epithelial cells of the testes in the vasectomized group were disorderly arranged and scattered. Significant atrophy and apoptosis were found in the endothelial cells, and a range of ultrastructural variations were observed in the cells of testes, epididymis, and vas deferens in vasectomized group. It was concluded that complete obstruction of the vas deferens as a traditional contraception method is not absolutely safe in terms of the reversal of fertility in the long run. Techniques of relieving the inner pressure in the vas deferens while maintaining the efficacy of male contraception need to be explored.

Objective To improve the different diagnosis between autoimmune pancreatitis(AIP) and pancreatic cancer(PC) by a retrospective analysis of clinical symptoms and serological features.Methods The analysis included 36 patients who had postoperative pathological,serological findings consistent with Asian AIP standards and 95 patients who had postoperative pathological consistent with PC pathological standards.All patients were admitted by the surgery department of our hospital from January,2003 to October,2011.A retrospective comparative analysis of the clinical manifestations,serology data of these AIP and PC patients was conducted.And summary the differential diagnosis characteristics of AIP and pancreatic cancer in the clinical symptoms,the serology.Results The features of different diagnosis:(1) The age of patients with PC was higher than AIP((60.9 ±9.0) years vs.(53.56 ± 14.6) years,t =3.48,P ＜0.05),and AIP preferred to male groups (x2 =2.88,P =0.09).(2) The clinical features of AlP and PC with the age characteristics were easily confused.Both clinical features were relatively typical in younger age which could be found earlier and relatively insidious in the older age which might be found with delay.(3) AIP were often complicated by biliary system inflammations(AIP =47.2％,PC =12.6％,x2 =18.12,P ＜ 0.05),while PC were usually complicated by the cysts in liver and kidney (PC =29.5％,AIP =0,x2 =13.50,P ＜ 0.05).(4) The high titer in CA199 had a higher value in the diagnosis of PC (concentration:group AIP =20.51 (9.55,86.5) kU/L,group PC =326.50 (94.38,10393.00) kU/L; positive rate:group AIP =35.70％ (10/28),group PC =86.70％ (65/75),P =0.000).The high titers in amylase (concentration:group AIP =103.50 (72.00,252.00) U/L,group PC =46.50 (21.65,96.90) U/L; positive rate:group AIP =45.00％ (9/20),group PC =19.40％ (7/36),P =0.043),lipase(concentration:group AIP =340.50(152.05,495.80) U/L,group PC =107.40(23.40,177.26) U/L,P =0.005 ; aspartate aminotransferase (positive rate:group AIP =75.00％ (27/36),group,PC =55.90％ (52/93),P =0.046) andγ-glutamyltranspeptidase (positive rate:group AIP =79.40％ (27/34),group PC =57.10％ (52/91),P =0.022) had higher values in the diagnosis of AIP.The significant increases in CA199 were not the basis which excluding AIP.Conclusion AIP as a unique type of chronic pancreatitis can be distinguished from PC on distinctive clinical,serological characteristics.

The effects of vasectomy on the reproductive organs in various species are controversial. This study investigated the morphological change and apoptosis of the testis, epididymis, and vas deferens in beagle dogs 12 months after vasectomy. The male beagles were divided into two groups: vasectomized and sham-operated groups (n=5 in each). Histopathological, ultrastructural, and TUNEL evaluation of the changes in the testis, epididymis, and ductus deferens of each animal were conducted 12 months after surgery. The mean lumen diameter, cellular thickness, mean interstitial distance, and lumen area fraction of each seminiferous tubule and ductus epididymis were measured by stereological analysis. The results showed that, compared with the sham-operated group, the seminiferous tubular epithelial cells of the testes in the vasectomized group were disorderly arranged and scattered. Significant atrophy and apoptosis were found in the endothelial cells, and a range of ultrastructural variations were observed in the cells of testes, epididymis, and vas deferens in vasectomized group. It was concluded that complete obstruction of the vas deferens as a traditional contraception method is not absolutely safe in terms of the reversal of fertility in the long run. Techniques of relieving the inner pressure in the vas deferens while maintaining the efficacy of male contraception need to be explored.
Animals ; Dogs ; Epididymis ; surgery ; Male ; Testis ; surgery ; Vasectomy ; adverse effects

e to selectively transport Na+ and glucose.

Objective:To investigate the influence and mechanism of micro RNA (miR)-373-3p in autophagy and sunitinib sensitivity of glioblastoma cells.Methods:U251 cells were routinely cultured in vitro; and some U251 cells were subjected to 50 μmol/L sunitinib treatment for 72 h to construct sunitinib-resistant U251 cell line. (1) Real-time reverse transcription quantitative PCR (RT-qPCR) was used to detect the miR-373-3p expression in U251 and sunitinib-resistant U251 cells. Sunitinib-resistant U251 cells were divided into blank control group, nonsense sequence group and miR-373-3p mimic group; cells in the latter 2 groups were transfected with nonsense sequence and miRNA-337-3p mimic, respectively; miR-373-3p expression was detected by RT-qPCR. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, CCK-8 assay was used to evaluate the cell viability; TUNEL was used to detect the apoptotic rate; immunofluorescent assay was used to detect the expression of microtubule-associated protein light chain 3 (LC3); Western blotting was used to detect the expressions of apoptosis- and autophagy-associated proteins. (2) The pGL3-autophagy-related gene 7 (ATG7) wild-type (WT) and pGL3-ATG7 mutant type (MUT) plasmids were established; dual-luciferase reporter system was used to detect the cell luciferase activity in the miR-373-3p mimic group and nonsense sequence group. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of ATG7 in the cells. (3) The sunitinib-resistant U251 cells were divided into blank control group, ATG7 negative control group, and ATG7 overexpression group; after each transfection, RT-qPCR and Western blotting were used to detect the ATG7 mRNA and protein expressions. U251 and sunitinib-resistant U251 cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, sunitinib-resistant U251+miR-373-3p mimic group, sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, and sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group; after each transfection, CCK-8 assay was used to evaluate the cell apoptosis, TUNEL was used to examine the apoptotic rate, and Western blotting was employed to detect the expressions of apoptosis- and autophagy-associated proteins. Results:(1) As compared with that in the U251 cells, miR-373-3p was lowly expressed in sunitinib-resistant U251 cells, with statistic difference ( P<0.05). As compared with that in the blank control group and nonsense sequence group, miR-373-3p expression was significantly elevated in the miR-373-3p mimic group ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had significantly increased cell viability, significantly decreased cell apoptotic rate, statistically increased B lymphocytoma-2 (Bcl-2) and Beclin 1 protein expressions, and significantly increased LC3II/LC3I values, significantly decreased Bcl-2 associated X protein (Bax) and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, and significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had increased number of fluorescent particles labeled with LC3 and enhanced fluorescent intensity; as compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had decreased number of fluorescent particles labeled with LC3 and reduced fluorescent intensity. (2) The luciferase activity of pGL3-ATG7 WT plasmids in the miR-373-3p mimic group was signficantly lower than that in nonsense sequence group ( P<0.05). As compared with those in the U251 group, ATG7 mRNA and protein expressions were both significantly increased in the sunitinib-resistant U251 group ( P<0.05); as compared with those in the sunitinib-resistant U251+nonsense sequence group, ATG7 mRNA and protein expressions were both significantly decreased in the sunitinib-resistant U251+miR-373-3p mimic group ( P<0.05). (3) As compared with the blank control group and ATG7 negative control group, the ATG7 overexpression group had significantly increased ATG7 mRNA and protein expressions ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions, and significantly increased cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, the sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group had significantly increased cell viability, significantly decreased apoptotic rate, statistically increased Bcl-2 and Beclin 1 protein expressions, significantly increased LC3II/LC3I values, significantly decreased Bax and p62 protein expressions, and significantly decreased cleaved Caspase3/Caspase 3 values ( P<0.05) Conclusion:MiR-373-3p can enhance sunitinib sensitivity by regulating autophagy in glioblastoma cells, whose mechanism might be related to targeting ATG7.