Abstract： Objective To investigate the effects of calcium preconditioning (CP) on immature myocardial cell apoptosis and apoptosisregulated proteins. Methods The experiment was carried out from June 2000 to December 2001 in the Renmin Hospital of Wuhan University. Twelve rabbits with the age of 1421 d and the weight of 230300 g were divided into 2 groups with 6 in each group by random digital table. For rabbits in the ischemia/reperfusion group (I/R group), after Langendorff models were routinely set up, KrebsHenseleit (KH) solution was perfused for 20 minutes and reperfused for 120 minutes after 45 minutes of ischemia. For rabbits in the CP group, after Langendorff models were established, KH solution was perfused for20 minutes, and 45 seconds’ noncalcium KH solution perfusion and 5 minutes’ KH solution perfusion were repeated 3 times before 45 minutes of ischemia and 120 minutes of reperfusion of KH solution. In situ apoptosis identification and semiquantitative analysis were used to detect the myocardial cell apoptosis; agarose gel electrophoresis was used to detect the nucleosomal ladder of DNA fragments; and the expression of bcl-2, bax and fas were detected with Western blot method. Results The apoptosis rate for the CP group was lower than that of the I/R group (4.53%±1.22% vs. 12.30%±2.12%,t=7.780, P=0.000). Nucleosomal ladder of DNA fragments of the CP group was lower than that of the I/R group (OD value: 56 460±1 640 vs. 135 212±3 370,t=51.460,P=0.000). The expression of bcl-2 in the I/R group was lower than that of the CP group (OD value: 13 217±1 770 vs. 31 790±1 018,t=22.280, P=0.000). The expression of bax (OD value: 30 176±1 025 vs. 7 954±730, t=43.260, P=0.000) and fas (OD value: 29 197±1 233 vs. 8 140±867, t=34.220, P=0.000) in the I/R group was higher than that of the CP group. Conclusion CP can affect the expression of myocardial bcl-2, bax, and fas, and decrease immature myocardial cell apoptosis.
Objective To investigate the effects of tight blood glucose control on the shortterm prognosis of the patients after heart valve replacement, in order to improve treatment effectiveness and lower postoperative complications. Methods A total of 240 patients including 150 males and 90 females underwent mitral valve replacement or mitral and aortic valve replacement were enrolled in this study from January 2007 to December 2008 at the cardiac surgery department of Renmin hospital of Wuhan university. The age of these patients ranged from 19 to 65 years old with an average age of 53.33 years. According to insulin administration time and blood glucose control level, they were randomly separated into two groups. In the experimental group, there were 121 patients who received continuous insulin infusion to maintain postoperative glucose level between 4.4 and 6.1 mmol/L, while 119 patients in the control group received insulin infusion when their glucose level went higher than 11.1 mmol/L to control the level between 6.1 and 11.1 mmol/L. Then the postoperative wound infection, malignant arrhythmia rate, the assisted ventilation time, intensive care unit(ICU) stay time and count of neutrophils were compared and analyzed. Results There was no hospitalized death in both groups. The rate of wound infection（3.31% vs.10.08%, χ2=4.430,P=0.035）, the assisted ventilation time（9.02±2.73 h vs. 10.01±3.58 h, t=2.280,P=0.024）, time for count of leukocytes to decrease to the normal level（11.04±3.16 d vs. 12.05±3.76 d, t=2.168,P=0.031）, average hospitalization time（13.49±3.81 d vs. 14.51±4.02 d,t=2.017,P=0.045）, and count of neutrophils on the third day（0.82±0.04 vs. 0.84±0.05, t=2.644,P=0.009） in the experimental group were significantly lower or shorter than those in the control group. But there was no significant difference between both groups in ICU stay time and the rate of malignant arrhythmia. Conclusion Tight blood glucose control can lower the rate of postoperative wound infection, shorten the assisted ventilation time and hospitalization time, and reduce the usage of antibiotics in patients after heart valve replacement. Accordingly, it can enhance the curative effect, reduce overall medical expenses, and improve prognosis.
Abstract： Objective To investigate the mechanism of protein kinase C(PKC) in immature myocardial ischemic preconditioning in order to further its clinical applicability. Methods Langendorff perfusion heart models of 24 rabbits were set up and they were randomly divided into 4 groups: ischemic reperfusion group (I/R group), myocardial ischemic preconditioning group (MIP group), chelerythrine group (CLT group) and protein kinase C group (PKC group). The emodynamics, biochemistry and myocardial ultrastructure were observed. Results The heart function recovery and myocardial water content in the MIP and the PKC groups were better than those of the I/R and the CLT groups (Plt;0.01). The adenosine triphosphate (ATP) content, superoxide dismutase activity, mitochondrial Ca2+-ATPase activity and synthesizing ATP activity of mitochondria in the MIP and the PKC groups were significantly higher than those of the I/R and the CLT groups (Plt;0.01). The dehydrogenase and creatine kinase leakage, malondialdehyde content, myocardial cell Ca2+ content and mitochondrial Ca2+ content in the MIP and the PKC groups were significantly lower than those of the I/R and the CLT groups (Plt;0.01). The myocardial ultrastructure injuries in the MIP and the PKC groups were less than that of the I/R and the CLT groups. Conclusion Myocardial ischemic preconditioning plays an important role in protecting immature myocardium, which is probably realized by the activation of PKC.
ObjectiveThis study is aimed to determine the expression of ubiquitin-specific peptidase 39 (USP39) protein in the colorectal cancer (CRC) tissues, and the effect of silencing USP39 gene on the cell growth and cell cycle distribution of CRC cells.Methods① The expressions of USP39 protein in CRC tissues and its paracancerous tissues were determined by immunohistochemical staining method. ② By lentiviral infection, Lv-shUSP39 (KD-1 and KD-2 group) and Lv-shCon (shCon group) were transferred into SW1116 and HCT116 cells, and cells of blank control group did not received any treatment (Con group). To determine the role of USP39 gene in cell growth, MTT assay was performed to draw growth curve, and cell cycle distribution of CRC cells in the 4 groups were determined by flow cytometer.Results① The expression of USP39 protein was higher in CRC tissues compared to adjacent tissues (P=0.007). ② For SW1116 and HCT116 cells, the cell proliferation ability of KD-1 and KD-2 groups were remarkably decreased than those in corresponding shCon and Con groups on 3, 4, and 5-day (P<0.05). ③ Flow cytometry assay showed that, the percentage of G0/G1 phase cells were decreased obviously (P<0.05), while increased significantly in percentage of G2/M phase and number of sub-G1 phase cells in KD-1 group compared with that in the Con group and shCon group of SW1116 and HCT116 cells (P<0.05).ConclusionsThe expression of USP39 protein is highly expressed in CRC tissues. Knockdowning of USP39 gene can inhibit cell proliferation and promote cell apoptosis.
Objective To investigate the short-term effectiveness of unilateral biportal endoscopic transforaminal lumbar interbody fusion (UBE-TLIF) in the treatment of Meyerding degree Ⅰ or Ⅱ single-segment lumbar spondylolisthesis. MethodsThe clinical data of 26 patients with Meyerding degree Ⅰ or Ⅱ single-segment lumbar spondylolisthesis treated with UBE-TLIF between January 2021 and August 2021 were retrospectively analyzed. Among them, there were 10 males and 16 females with a mean age of 61.5 years (range, 35-76 years). The lesion segment included L3, 4 in 2 cases, L4, 5 in 18 cases, and L5, S1 in 6 cases. There were 17 cases of degenerative spondylolisthesis and 9 cases of isthmic spondylolisthesis; according to the Meyerding classification of spondylolisthesis, 19 cases were grade Ⅰ and 7 cases were grade Ⅱ. Twenty-one cases were complicated with lumbar disc herniation and spinal stenosis and 5 cases with lumbar spinal stenosis. The operation time, hospitalization stay, complications, hemoglobin (Hb) and serum creatine kinase (CK) levels before operation and at 1 day after operation were recorded; lumbar lordosis angle changes and postoperative spondylolisthesis reduction were evaluated by lumbar anteroposterior and lateral X-ray films before operation and at last follow-up; visual analogue scale (VAS) score was used to evaluate the low back pain and leg pain before operation, at 2 days, 1 week, 2 weeks after operation, and at last follow-up; Oswestry disability index (ODI) was used to evaluate the functional recovery of the patients before operation and at last follow-up. ResultsThe operation was successfully completed in all 26 patients, with an average operation time of 181.9 minutes (range, 130-224 minutes) and an average hospitalization stay of 6.3 days (range, 3-9 days). Hb levels were significantly lower and serum CK levels were significantly higher at 1 day after operation when compared with those before operation (t=7.594, P<0.001; t=–15.647, P<0.001). No serious complication occurred during and after operation. CT examination at 3 days after operation showed that the percutaneous screw was not in good position in 1 case, and nerve paralysis (pain, numbness) occurred in 2 cases after operation, which were improved within 2 weeks after operation. All the 26 patients were followed up 6-11 months, with an average of 8.7 months. Complete reduction (the slippage reduction rate was 100%) was achieved in 24 patients (92.3%), and partial reduction (the slippage reduction rate was 87.5%) in 2 patients (7.7%). During the follow-up, there was no complication such as incision infection, fusion Cage subsidence or displacement, and internal fixator loosening. The VAS scores of low back pain and leg pain significantly improved at each time point after operation when compared with those before operation (P<0.05); there was no significant difference in the VAS scores of low back pain and leg pain between at 2 days and 1 week after operation, the VAS scores of low back pain between at 1 week and 2 weeks after operation, and the VAS scores of leg pain between at 2 weeks after operation and last follow-up (P>0.05); but there was significant difference between the other time points after operation (P<0.05). ODI and lumbar lordosis angle significantly improved at last follow-up (P<0.05). Conclusion UBE-TLIF provides favorable short-term effectiveness and obvious advantages of minimally invasive in the treatment of Meyerding degree Ⅰ or Ⅱ single-segment lumbar spondylolisthesis. However, the safety and long-term effectiveness need to be further studied.
ObjectiveTo analyze the status of applying programmed death-1 (PD-1) / programmed death-ligand 1 (PD-L1) inhibitors combined with vascular endothelial growth factor (VEGF) / vascular endothelial growth factor receptor (VEGFR) inhibitors in advanced refractory colorectal cancer. MethodThe relevant literature on domestic and foreign research in recent years was summarized. ResultsThe discovery of immune checkpoint PD-1/PD-L1 and the clinical application of related drugs had changed the treatment pattern of advanced solid tumors, but PD-1/PD-L1 inhibitors had a poor efficacy in the mismatch repair prodicient tumors, and most advanced colorectal cancer belonged to this type. The combination of PD-1/PD-L1 inhibitors and VEGF/VEGFR inhibitors could enhance the therapeutic effect in the advanced refractory colorectal cancer, and their interaction mechanisms and clinical efficacy were continuously being proven. ConclusionsThe combination of PD-1/PD-L1 inhibitors and VEGF/VEGFR inhibitors is a promising treatment strategy for advanced refractory colorectal cancer. More studies need to be further clarified its efficacy.
ObjectiveGelatin methacryloyl (GelMA)/hyaluronic acid methacryloyl (HAMA)/chitosan oligosaccharide (COS) hydrogel was used to construct islet biomimetic microenvironment, and to explore the improvement effect of GelMA/HAMA/COS on islet activity and function under hypoxia. Methods Islets cultured on the tissue culture plate was set as the control group, on the GelMA/HAMA/COS hydrogel with COS concentrations of 0, 1, 5, 10, and 20 mg/mL respectively as the experimental groups. Scanning electron microscopy was used to observe the microscopic morphology, rheometer test to evaluate the gel-forming properties, contact angle to detect the hydrophilicity, and the biocompatibility was evaluated by the scaffold extract to L929 cells [using cell counting kit 8 (CCK-8) assay]. The islets were extracted from the pancreas of 8-week-old Sprague Dawley rats and the islet purity and function were identified by dithizone staining and glucose-stimulated insulin secretion (GSIS) assays, respectively. Islets were cultured under hypoxia (1%O2) for 24, 48, and 72 hours, respectively. Calcein-acetyl methyl/propidium iodide (Calcein-AM/PI) staining was used to evaluate the effect of hypoxia on islet viability. Islets were cultured in GelMA/HAMA/COS hydrogels with different COS concentrations for 48 hours, and the reactive oxygen species kits were used to evaluate the antagonism of COS against islet reactive oxygen species production under normoxia (20%O2) and hypoxia (1%O2) conditions. Calcein-AM/PI staining was used to evaluate the effect of COS on islet activity under hypoxia (1%O2) conditions. Islets were cultured in tissue culture plates (group A), GelMA/HAMA hydrogels (group B), and GelMA/HAMA/COS hydrogels (group C) for 48 hours, respectively. Immunofluorescence and GSIS assays were used to evaluate the effect of COS on islet activity under hypoxia (1%O2) conditions, respectively. Results GelMA/HAMA/COS hydrogel had a porous structure, the rheometer test showed that it had good gel-forming properties, and the contact angle test showed good hydrophilicity. CCK-8 assay showed that the hydrogel in each group had good biocompatibility. The isolated rat islets were almost round, with high islet purity and insulin secretion ability. Islets were treated with hypoxia for 24, 48, and 72 hours, Calcein-AM/PI staining showed that the number of dead cells gradually increased with time, which were significantly higher than those in the non-hypoxia-treated group (P<0.001). Reactive oxygen staining showed that GelMA/HAMA/COS hydrogels with different COS concentrations could antagonize the production of reactive oxygen under normal oxygen and hypoxia conditions, and this ability was positively correlated with COS concentration. Calcein-AM/PI staining indicated that GelMA/HAMA/COS hydrogels with different COS concentrations could improve islet viability under hypoxia conditions, and cell viability was positively correlated with COS concentration. Immunofluorescence staining showed that GelMA/HAMA/COS hydrogel could promote the expression of islet function-related genes under hypoxia conditions. GSIS assay results showed that the insulin secretion of islets in hypoxia condition of group C was significantly higher than that of groups B and C (P<0.05). Conclusion GelMA/HAMA/COS hydrogel has good biocompatibility, promotes islet survival and function by inhibiting reactive oxygen species, and is an ideal carrier for building islet biomimetic microenvironment for islet culture and transplantation.
Objective To investigate the clinical applications of plasma shock wave lithotripsy （PSWL） in the treatment of calculi via choledochoscope. Methods Between 2004 and 2009, 56 huge residual calculi (54 cases) were underwent PSWL via choledochoscope treatment in our hospital. Calculi size: diameter ≤10 mm in 9 calculi, 10 mm lt;diameter ≤15 mm in 24 calculi, 15 mm lt;diameter ≤20 mm in 17 calculi, and diameter gt; 20 mm in 6 calculi. Twenty four cases of these 54 patients had bile duct stricture. Procedure: Firstly found the calculus, and then the lithotriptor wire was introduced to the place 0.5-1.0 mm far away from the calculus surface through choledochoscope channel, and powered on, fired, destructed. Results Forty-eight cases of 54 patients were cured by PSWL with 1 times, 2 cases with 2 times, 2 cases with 3 times, and 2 cases with 4 times. Shock 4-300 times were exported per PSWL, with an average of 65 times. Crushing each stone needed shock for 4-680 times, with an average of 77 times. The calculi were ruptured in different degrees by PSWL, of which 20 cases were completely ruptured. All calculi were completely extracted. Except one case with bile duct stricture was found a small amount of bile duct bleeding during operation, all other patients had no operation complications. Conclusion PSWL technique plays an important role in lithotripsy. It is easy to deal with intrahepatic impacted calculi by PSWL, especially the pigment stones with rough surface.
Objective To investigate the feasibility and efficacy of transcatheter directed thrombolysis （TDT） approach in treatment for deep venous thrombosis （DVT） of lower limbs and as compared with trans-dorsal pedis vein thrombolysis （TPVT） approach. Methods The clinical data of 437 patients with acute DVT （184 males and 253 females） at the age of （43±12） years （range 19-76 years） from July 2008 to January 2012 in the First Affiliated Hospital of Zhengzhou University were analyzed retrospectively. Patients in the group TDT received TDT were 293 cases， 32 inferior vena cava filters were implanted. Patients in the group TPVT received TPVT were 65 cases， 4 inferior vena cava filters were implanted. Results The resolution time of thrombus in the group TDT was shorter than that in the group TPVT （6 d versus 9 d， P＜0.05）. The circumference difference of leg or upper leg before and after treatment in the TDT group was significantly greater than that in the TPVT group （P＜0.05）. The rate of venous patency was （65.2±15.4）% and preservation rate of valvular function was （78.2±12.6）% in the group TDT， and which was （63.8±16.3）% and （91.1±10.7）% in the group TPVT， respectively. The differences of venous patency rate was not statistically significant（P＞0.05） between two groups， but the prevervation rate of valvular function was significant difference （P＜0.05） .Hematomas in 3 cases and gross hematuria in 4 cases were observed， and displacement of inferior vena cava filter occurred in 1 patient in the group TDT. The gums bleed or gross hematuria in 5 cases were observed in the group TPVT. Conclusions Both TDT and TPVT can effectively relieve symptoms. TDT can shorten the course of disease， but itincreases functional damage of the deep vein valvular.