Objective To study the mechanisms of increasing amyloid [β1-42 (Aβ42) level by presenilin-1 (PS-1) mutation of V97L.Methods Expression level of neprilysin (NEP) in nontransfected, or mock vector, wild type (wt) PS-1 and V97L-PS-1-transfected SH-SY5Y cells were assessed by reverse transcriptional-polymerase chain reaction (RT-PCR) and Western blot.The level of Aβ42 was also assessed by enzyme linked immunosorbent assay (ELISA).Results The mRNA level of NEP was decreased significantly in PS-1 V97L transfected cells (0.650 ± 0.010) compared to that in normal cells (1.090 ± 0.015), wt PS-1 gene (1.040 ± 0.021) and mock (1.080 ± 0.020) stably transfected cell lines (t = 9.236, 10.452, 5.678; all P < 0.01).Although there was a decreased tendency in the protein expression of NEP in PS-Ⅰ V97L transfected cells (1.000 ± 0.126)compared to that in normal cells (1.020 ± 0.110), wt PS-1 gene (1.040 ±0.110), mock (2.130 ±0.130) stably transfected cell lines, nosignificant differences were found.The expression of Aβ42 was increased significantly in PS-1 V97L transfected cells compared to other cell lines (t = 2.109, 3.355, 3.976; all P < 0.01).Conclusions PS-1 V97L mutation can decrease the transcription of NEP, however, in contrast, this mutation also resulted in increased Aβ42.The mechanism needs further analysis.

Objective To study the type variation of microglial activation in spinal dorsal horn of rats after sciatic nerve injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into the control and experimental groups, 24 rats in each group.The experimental group underwent ligation of sciatic nerve trunk to generate nerve injury in the rats.The pain behavior in the rats was measured at the 1th, 7th and 14th postoperative days, and the changes of microglial activation in the rat lumbar spinal cord dorsal horn was detected by immunofluorescence staining.qRT-PCR assay was used to validate the activation trends of M1 and M2 types of microglia cells.Results No significant changes were found in the microglial cells in the spinal cord dorsal horn of rats in the sham-operation group during 14 days after operation.In the sciatic nerve ligation group at 1 day after operation, no significant change was observed in the number of microglial cells, but the expression of marker of M1 microglia was significantly increased.At 7 and 14 days after operation, the number of microglial cells and the expression of M1 microglia marker in the spinal cord dorsal horn were increased significantly.Conclusions Microglia activation in the spinal dorsal horn starts at the first day after sciatic nerve injury, and lasts at least for two weeks after the operation.M1 microglia activation dominates during this period.

Objective To investigate the effect of surfactant protein A (SP-A) on the production of MIP-2 and binding activity of NF-?B in rat tubular epithelial cells, and evaluate its possible role in renal inflammation. Methods Confluent cultures of NRK-52E cells (a renal tubular epithelial cell line of rat origin) were pretreated with various concentrations of SP-A(0 to 80 ?g/ml) and stimulated by lipopolysaccharide (LPS) (10 ?g/ml) with 2% serum. MIP-2 expression was measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The effect of SP-A on NF-?B binding activity was assessed by electrophoretic mobility shift assay (EMSA). Results MIP-2 mRNA and protein was expressed and up-regulated in NRK-52E cells stimulated by LPS. The expression of MIP-2 was down-regulated by SP-A. NF-?B binding activity was inhibited by SP-A in a concentration-dependent manner. Conclusion SP-A binding activity and down-regulates the expression of MIP-2 in renal tubular epithelial cells, which may play an important role in the modulation of renal tissue inflammation.

Objective To evaluate the effect of ALD on podocyte apoptosis and the possible roles of Akt in ALD-induced apoptosis. Methods The cultured rat podocytes were incubated with increasing concentrations of ALD (10-9~10-5 mol/L) for variable time periods. Apoptosis was evaluated by cell nucleus staining and flow cytometry. RT-PCR was used to examine the expression of mineralocorticoid receptor (MR)and 11 Beta-hydroxysteroid dehydrogenase type 2 (11?-HSD2) mRNA in podocyte. Activation of Akt/PKB was evaluated by performing Akt kinase assay. Results ALD induced podocyte apoptosis in a dose- and time-dependent manner. The proapoptotic effect was attenuated by the presence of spironolactone (10-7mol/L). The expression of MR and 11P-HSD2 mRNA was demonstrated in the podocytes by RT-PCR. ALD also inhibited the activity of Akt in a dose-dependent manner, but the inhibitory effect was significantly ameliorated by the presence of spironolactone. The activity of Akt was negatively correlated with podocyte apoptosis. Conclusion ALD induces apoptosis in rat podocytes through the signaling mechanism by which Akt is inhibited.

Objective To explore the pattern of lymph node metastasis in patitsen with squamous cell carcinoma of the thoracic esophagus and its significance in lymphadenectomy.Methods The clinical data of 230 patients who received radical esophagectomy with three-field lymphadenectomy was analyzed.The metastatic sites of lymph nodes were correlated with tumor location by chi-square test.Logistic regression was used to analyze the relationship between clinic pathoingical factors and lymph node metastasis.Results Lymph node metastases were found in 133 of the 230 patients(57.8%).The average number of resected lymph nodes was 25.3? 11.4 per patient(range 11～71).The rates of lymph node metastasis were 41.6%,19.44%and 8.3%in the neck,thoracic medi- astinum and abdominal cavity for patients with upper thoracic esophageal carcinoma,33.3%,34.7%and 14%for patients with mid- die thoracic esophageal carcinoma and 36.4 %,34.1%and 43.2 %for patients with lower thoracic esophageal carcinoma.No signifi- cant difference in cervical or thoracic metastatic rate was observed among upper,middle and lower thoracic carcinoma.The difference in lymph node metastatic rate for nodes in the abdominal cavity was significant among upper,middle and lower thoracic carcinoma. The lower thoracic esophageal cancers were more likely to metastasize to the abdominal cavity.Logistic-regression showed depth of tu- mor invasion and angiolymphatic invasion were factors influencing lymph node metastasis.Conclusion Cervical and mediastinal node dissection should be performed independently from tumor location.Abdominal node dissection should be conducted more vigorously for lower thoracic esophageal cancer than of other locations.Patients with greater tumor grade,depth of tumor invasion and angiolymphatic invasion were more prone to develop lymph node metastasis.

Objective To investigate the relationship between interleukin-17 (IL-17) and vitiligo.Methods Totally,32 vitiligo patients and 30 healthy human controls were enrolled in this study.Blood samples were collected from all the subjects,and enzyme-linked immunosorbent assay (ELISA) was performed to determine the plasma levels of IL-17.The relationship of plasma IL-17 levels with disease stage,clinical course and lesion area was assessed.Results The plasma levels of IL-17 were significantly higher in the patients with progressive and stable vitiligo than in the healthy controls (both P ＜ 0.05),and higher in the patients with progressive vitiligo than in those with stable vitiligo (P ＜ 0.05).Moreover,the plasma levels of IL-17 were positively correlated with the area of vitiligo lesions (r =0.456,P ＜ 0.05),but unrelated to the clinical course of vitiligo (r =0.239,P ＞ 0.05).Conclusion IL-17 may play a certain role in the occurrence and development of vitiligo.

OBJECTIVEThis study measures the glutaredoxin (Grx) gene and protein expression in umbilical vein endothelial cells upon exposure to Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS). The involvement of the Akt-signaling pathway is also determined.

METHODSEA-hy926 cells were pretreated with 1,000 ng · mL⁻¹ P. gingivalis LPS for 4, 12, 18, and 24 h, and then real-time reverse transcription polymerase chain reaction was employed to detect Grx1 expression. The effect of Grx on Akt activity was investigated using Western blot for the control, LPS (1,000 ng · mL⁻¹ LPS), and carmus- tine (BCNU) groups (1,000 ng · mL⁻¹ LPS, and the EA-hy926 cells were pretreated with 25 μmol · ml⁻¹ BCNU for 30 min).

RESULTSGene expression of Grx1 significantly increased in LPS group compared with that in the control group. The Grx1 expression reached the peak level in 12 h, and the variation between the expression in 4 and 12 h was significant (P < 0.05). After 12 h, the protein levels of Grx and phosphorylated-Akt (p-Akt) significantly increased in the LPS group (P < 0.05), whereas the BCNU group showed a considerable decrease in both Grx and p-Akt expression levels (P < 0.05). Moreover, a slight difference was observed in the total Akt protein levels in the three groups (P > 0.05).

CONCLUSIONGrx expression increased upon exposure of EA-hy926 cells to the LPS. Akt activity could be inhibited by BCNU (a Grx inhibitor), which indicated that Akt might act as a downstream regulator of Grx.

Endothelial Cells ; Glutaredoxins ; genetics ; Humans ; Lipopolysaccharides ; pharmacology ; Oxidative Stress ; drug effects ; Phosphorylation ; Porphyromonas gingivalis ; pathogenicity ; Proto-Oncogene Proteins c-akt ; drug effects ; Signal Transduction ; drug effects ; Umbilical Veins

Objective To investigate the possible protective effect of neurotropin on the corresponding spinal cord motoneuron after sciatic nerve injury early in rats.Methods 108 male Sprague Dawley rats were divided randomly into Groups A,B and C with 36 animals in each group,and all animals were cut both side of sciatic nerve,instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve.Group A was control group.Group B was treated with neurotropin and Group C was treated with normal saline after operation.The rats were sacrificed on 1st d,4th d,9th d,14th d,21st d and 28th d after operation.The corresponding spinal cord segments(L4～L6)were gotten to make Bcl-2 and Bax immunohistochemical staining and TUNEL staining.Results The values of Bcl-2/Bax on 9th d,14th d and 21st d after operation of Group B were different from that of Group A and Group C.The number of positive cells stained by TUNEL staining on 9th d was significantly different among Group A,Group B and Group C(v<0.05).The expression of positive TUNEL staining cells reached a peak on 14th d after operation,the number of Group B was less than that of Group A and Group C.Conclusion After instantly make the operation of end-to-end anastomose of epineurium for abscised sciatic nerve,neurotropin can protect the spinal cord motoneurons to avoid apoptosis excessively in some extent.

Objective:To study the indications and clinical efficacy of video assisted small incision in treatment of infected pancreatic necrosis.Methods:A retrospective study was conducted on 27 patients with infected pancreatic necrosis treated by video assisted small incision at the Department of Pancreatic and Splenic Surgery, Hunan Provincial People's Hospital, from January 2018 to December 2019. The surgical approach, operation time, intraoperative blood loss, postoperative hospital stay and complications were analysed. Postoperative follow-up was carried out at outpatients’ clinic, and the patient's time to full recovery and long-term complications were studied.Results:The 27 patients successfully underwent the operations. There were 22 males and 5 females, aged (50.6±6.2) years. The treatment results were analyzed according to the different surgical approaches: the retroperitoneal approach group ( n=4); the omental sac approach group (n=14); the intercostal space approach group ( n=2); and the combined approach group ( n=7). The operation time was (85.3±31.6)min. The intraoperative blood loss was 65.0(45.2, 121.4)ml. The postoperative hospital stay was 23.0(12.5, 36.1)days. The incidence of complications (Clavien-Dindo grade Ⅲ and above) was 14.8%(4/27). There were 2 patients in this study who were admitted to the intensive care unit due to postoperative hemorrhage: 1 patient responded well to conservative treatment and the remaining patient required interventional treatment. Another patient because of poor results, underwent debridement by laparotomy 2 weeks after the operation. There was 1 patient who developed grade C pancreatic fistula which was cured by surgical treatment 6 months later. On follow-up, 2 patients developed colonic fistula 2 weeks after surgery and 2 patients gastric fistula 1 week and 3 weeks after surgery. These patients responded to conservative treatment. Conclusion:With proper case selection, video assisted small incision could safely and effectively be used to treat infected pancreatic necrosis.

In recent years, with the application and exploration of fluorescence imaging technology in pancreatic surgery, it has brought new changes and new hope for the intraoperative treatment of pancreatic diseases. Intraoperative fluorescence imaging with indocyanine green (ICG) as probe is increasingly applied in identify neoplasms, real-time margin assessment, tissue perfusion, lymph node detection, anastomotic leakage and so on, which provides a new means for intraoperative decision-making. This article reviewed the progress in the application of ICG fluorescence imaging in pancreatic surgery.