Objective To discuss the efficacy and cosmetic effect of laparoscopic thyroidectomy for patients with thyroid diseases. Methods The clinical data of 160 patients who underwent laparoscopic thyroidectomy through the anterior chest approach were analyzed. The operation was performed under a pressure of 8 mm Hg within the surgical space at the neck. After the thyroid was separated completely, the lesions were resected using an ultrasound knife. Results Laparoscopic thyroidectomy was completed in 157 of the patients including 6 cases of papillary adenocarcinoma. The other 3 patients were converted to open surgery because of hyperthyroidisms complicated with intraoperative hemorrhage (1) or thyroid carcinoma complicated with lymph node metastasis (2). No severe complications involving the trachea and parathyroid occurred in this series. One patient with thyroid carcinoma developed transient hoarseness after the operation; one patient with nodular goiter had postoperative subcutaneous hemorrhage and hydrops; both of them were cured spontaneously. Postoperative hospital stay ranged from 3 to 5 days (mean, 4 days). The patients were followed up for 3 to 24 months with a mean of 6.8 months, during which none of them had recurrence. All the patients were satisfied with cosmetic outcomes of the surgery. Conclusion Laparoscopic thyroidectomy via the anterior chest approach is safe and feasible for patients with thyroid diseases with good cosmetic results.

Objective To identify a detailed biological characterization of mesenchymal stem cells (MSCs) isolated from hu‐man umbilical cord(UC) tissue regarding their morphology ,immunophenotype ,purity and proliferative capacity and establish a rea‐sonably cultured and amplified system .Methods After stripping off arteries and veins ,the remaining parts of umbilical cord were cut into 1 mm3 small sections and cultured with DMEM/F12 containing 10% fetal bovine serum .Adhere cells were obtained and the morphology of the cells was observed under inverted phase contrast microscope .The growth curves of them were drawn by CCK‐8 and the cell cycle and surface antigens (CD29 ,CD73 ,CD90 ,CD105 ,CD31 ,CD14 ,CD34 ,CD45 ,CD11b ,HLA‐DR) were detected by flow cytometry .Results Seven to ten days after primary culture ,adhere cells came out of fragments .The MSCs harvested were a high purity and mainly presented as a fibroblast‐like morphology .UC‐MSCs had a strong ability of proliferation through the cell growth curve .The special surface antigens CD29 ,CD73 ,CD90 ,CD105 were positive expression ,while CD31 ,CD14 ,CD34 ,CD45 , CD11b ,HLA‐DR were negative .More than 80% cells of MSCs were found at G0/G1 phase .Conclusion Human UC‐MSCs could be cultured and proliferated in vitro .

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

ObjectiveTo explore the feasibility, safety and clinical value of laparoscopic Rouxen-Y cystojejunostomy in the treatment of pancreatic pseudocyst. Method Four patients with pancreatic pseudocyst received totally laparoscopic pancreatic pseudocystojejunostomy. The data on intraoperative bleeding, operative time, postoperative time to get out of bed, time of first flatus/bowel motion, complication and duration of hospital stay were collected and analyzed retrospectively. ResultsAll operations were carried out successfully with laparoscopic surgery. The mean operative time was 90 min. The average intraoperative blood loss was 40 ml. The mean postoperative time to get out of bed was 1.5 d, and the mean time of first flatus/bowel motion was 2. 3 d. All patients recovered smoothly without any pancreatic fistula. The average hospital stay was 7 days. Fever, pancreatitis,adhesive intestinal obstruction and other complications did not occur. ConclusionsTotally laparoscopic Roux-en-Y pancreatic pseudocystojejunostomy was an efficacious, safe, and minimally invasive procedure.

Objective To investigate the expression of pSTAT5 in 7 pancreatic carcinoma cell lines,and the change of expression of pSTAT5 in pancreatic carcinoma cells SW1990 after growth hormone (GH) treatment, and explore its molecular mechanism. Methods Human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, AsPc, P3, PANC1) were cultured in vitro, and Western blotting was used to detect the expression of pSTAT5 in these cell lines. SW1990 in exponential growth phase was collected and nude Balb/c mice were inoculated with SW1990 cells. When tumors became palpable after inoculation, mice (normal saline group). 1 h, 2 h and 24 h after the last dose of GH treatment, the mice were sacrificed.Western blotting was used to detect the expression of pSTAT5 in SW1990 and inoculation tumor cells after GH injection. Results Positive expression of pSTAT5 was observed in all human pancreatic carcinoma cell lines (SW1990, Cap-1, Colo, Mia, Aspc, P3, PANC1). 5 minutes after GH (50 ng/ml) stimulation, the expression of pSTAT5 in SW1990 was 0.57 ±0.05, which was significantly increased; and it reached 0.64 ±0.04 at 10 minutes, then decreased to 0.39 ±0.03 at 15 minutes, however, it remained higher than that in the control group at 1 h (0.33 ± 0.02 vs 0.25 ± 0.06), and its expression at 2 h was 0.26 ± 0.03 and returned to the normal level. The expression of pSTAT5 in xenograft was not significantly changed. Conclusions GH could rapidly up-regulate the expression of pSTAT5 in SW1990 but the effect lasted for a relatively short period. GH had no significant effect on the expression of pSTAT5 in xenograft.

Objective To explore the feasibility and safety of cotransplantation of autologous bone marrow-derived mesenehymal stem cells (MSCs) and peripheral blood stem cells in hematological malignant diseases and to observe its effect on hematopoietic reconstruction after cotransplantation. Methods Adult human MSCs were isolated from the healthy bone marrow of the patient himself with Percoll (1. 073 g/ml) and cultured in Dulbecco's modified Eagle's medium with low glucose containing 10% AB type human serum. After conditioning regimen of high-dose chemotherapy and radiotherapy, cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells was done in five patients with hematological malignant diseases. Results The process of the infusion was safe and there were no adverse reactions or other toxicities related to the infustion of MSCs. The median time to achieve neutrophil counts greater than 0. 5 × 109/L was 9.4 days ( ranging from 8 to 11 days) after cotransplantation and platelet counts greater than 20 × 109/L 12. 2 days (ranging from 10 to 14 days). Conclusion Cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells in hematological malignant diseases is feasible and safe. The rapid hematopoietic reconstruction after cotransplantation shows that MSCs have an effect on hematopeiesis, but the mechanism is still to be investigated.

Objective To investigate the effect of GH on proliferation of pancreatic cancer cells and observe the features of IGF-IGFBP3 pathway in the host after GH administration. Methods Pancreatic cancer cells (SW-1990,PANC-1 and P3) during exponential growth stage were harvested and cultured in medium containing growth hormone (50 ng/ml). After 24, 48 and 72 hours, cells were counted using a Coulter Counter. Thirty-five Athymic nude Balb/c mice were inoculated with SW-1990cells. When tumors became palpable after inoculation, animals were randomized to receive GH points (1 h, 2 h, 6 h, 24 after the last injection), plasma samples were gathered for subsequent ELISA determination and liver was rapidly incised for immune blotting analysis. Results The results revealed that GH stimulated cell growth in vitro. GH elevated levels of IGF-Ⅰ , Ⅱ at the 1st , 2nd , 6th hour after the last injection. GH augmented the expression of IGFBP3 in the liver of the host in vivo (1 h, 2 h, 6 h, 24 h, respectively). Conclusion Such proteins as IGF- Ⅰ and Ⅱ might be associated with mechanism of last effect of GH on tumor host. The up-regulation of IGFBP3 by GH administration in the host may help to explain the phenomena that GH doesn't accelerate growth of pancreatic tumor in vivo.

Objective To investigate the protection effect of cyclosporine A on spatial memory following chronic cerebral hypoperfusion in rats and its possible mechanism.Me,ods Sixty SD rats were randomly divided into sham operation,vehicle,low-dose cyclosporine A,medium-dose cyclosporine A,and high-dose cyclosporine A groups.A chronic cerebral hypoperfusion model was prepared by permanent bilateral ligation of bilateral common carotid arteries.From 46 days after modeling,olive oil 1 ml/d was used for intragastric administration in the sham-operation group and the vehicle group.Cyclosporine A 3 mg/kg,6 mg/kg,and 12 mg/kg were administrated intragastrically in the low-dose,medium-dose and high-dose cyclosporine A groups,respectively,once a day for 14 days.The spatial memory was assessed using Morris water maze test.Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of NADPH oxidase 4 (NOX4) mRNA in the cerebral cortex.Immunohistochemical staining and Western blot were used to detect the expression of NOX4 protein in the cerebral cortex.Results The Morris water maze test showed that the escape latencies in all cyclosporine A groups were significantly shorter than the vehicle group (all P ＜0.05).Immunohistochemical staining showed that the percentages of the NOX4-positive cells in the sham-operation,vehicle,low-dose,medium-dose,and high-dose cyclosporine A groups were4.43％ ±0.37％,37.44％ ±4.76％,18.05％ ±2.91％,12.51％ ±3.4％,and 11.06％ ±1.74％,respectively (F =262.021,P ＜ 0.001),the vehicle group was significantly higher than the sham-operation group (P ＜ 0.01),and all cyclosporine A groups were significantly less than the vehicle group (all P ＜ 0.01).RT-PCR showed that the expression levels of NOX4 mRNA in cerebral cortex in the sham-operation,vehicle,low-dose,medium-dose,and high-dose cyclosporine A groups were 0.36 ± 0.03,1.04 ± 0.04,0.58 ± 0.02,0.49 ± 0.01,and 0.40 ± 0.02,respectively (F =1 324.941,P ＜ 0.001),all cyclosporine A groups were significantly lower than the vehicle group (all P ＜ 0.01).Western blot showed that the expression levels of NOX4 protein in cerebral cortex in the sham-operation,vehicle,low-dose,medium-dose,and high-dose cyclosporine A groups were 0.02 ± 0.01,0.27 ± 0.04,0.09 ± 0.02,0.06 ± 0.02,and 0.06 ± 0.01,respectively (F =222.692,P ＜ 0.001),all cyclosporine A groups was significantly lower than the vehicle group (all P＜0.01).Conclusion Cyclosporine A may improve spatial memory following chronic cerebral hypoperfusion in rats by down-regulation of NOX4.

BACKGROUND:Studies have shown that paeoniflorin functions as replenishing blood and treatment of autoimmune diseases, and bone marrow mesenchymal stem cels also play an important role in the body’s blood and immune function. However, paeoniflorin effects on bone marrow mesenchymal stem cel proliferation and cytokine secretion and expression are rarely reported. OBJECTIVE:To investigate the effect of paeoniflorin on proliferation of bone marrow mesenchymal stem cels and the expression of interleukin-6. METHODS:Human bone marrow mesenchymal stem cels were separated and culturedin vitro by density gradient centrifugation combined with attachment method. The biological characteristics of bone marrow mesenchymal stem cels were identified by flow cytometry and osteogenic/adipogenic induction. The proliferation of bone marrow mesenchymal stem cels under different concentrations of paeoniflorin was detected by MTT method. The mRNA expression and secretion of interleukin-6 in the supernatant of bone marrow mesenchymal stem cels were detected by RT-PCR and ELISA, respectively. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated successfuly and had osteogenic and adipogenic differentiation potential. Compared with the controlgroup, 2 μmol/L and 10 μmol/L paeoniflorin significantly promoted the proliferation of bone marrow mesenchymal stem cels. 10 μmol/L paeoniflorin could significantly decrease the proportion of bone marrow mesenchymal stem cels in G0/G1 phase and increase this proportion in S phase. Compared with the control group, the experimental group could significantly increase the secretion and mRNA expression of interleukin-6 (P < 0.01). It is concluded that paeoniflorin at certain concentrations can obviously promote the proliferation of bone marrow mesenchymal stem cels, and increase the expression and secretion of interleukin-6.