【Abstract】ObjectiveTo establish an efficient, effective hepatocyte isolation technique in order to increase cell production and decrease the prime cost. Methods The inferior vena cava below diaphragm was dissected and ligatured, and the inferior vena cava below liver was separated. Subsequently, the liver was perfused with EGTA through the portal vein while the inferior vena cava below liver was opened, and then the liver was harvested. The liver tissue was cut into 1 mm×1 mm×1 mm and digested at 37 ℃ water bath with Ⅳ collagenase for 30-40 minutes, then the hepatocytes were purified and cultured in CO2 incubator. The production and function of hepatocytes were assessed. ResultsThe isolated hepatocytes using this technique were more than 95% among the all isolated cells. No statistic difference was found in cell production and cell function comparing with traditional technique. But this technique was simplified and more economically. ConclusionThis modified hepatocyte isolation technique is efficient and effective. It can ensure the amount of production and purity of hepatocytes.
【Abstract】ObjectiveTo investigate the causes of death in recipient rats after modified orthotopic liver transplantations and the countermeasure.MethodsTwo hundred and ten orthotopic liver transplantations were performed in Wistar and SD rats weighing 250～300 g through modified Kamada’s cuff technique, and to investigate the causes of death and preventive measures. ResultsIt took about (41±5) min, (13±2) min and (45±5) min for donor operation，donated liver trimming and recipient operation respectively. The anhepatic phase was (20±5) min. The 4week survival after transplantation was 90.5%. Nineteen rats died during and after transplantation. The causes of death included: bleeding of suprahepatic inferior vena cava (n=5 ), infrahepatic vena cava thrombosis (n=2), biliary obstruction (n=3 ), portal vein constriction and thrombosis (n=2 ), liver injury(n=1), bleeding of left subphrenic vein (n=2 ), infection (n=2 ), excessively deep anesthesia (n=1), and respiratory failure (n=1). ConclusionThe sophisticated microsurgical technique and the delicate surgical manipulation are very important in preventing operative complications, and the short anhepatic phase of recipient is the key to animal survival, and proper anesthesia is also important to the success of operations. Improving the condition of operation and reasonable use of antibiotics are necessary in preventing infections.
ObjectiveTo investigate the effect of multidisciplinary team (MDT) on perioperative complications and clinical efficacy of patients who were receiver radical operation to treat lung cancer by video-assisted thoracoscopic surgery (VATS). MethodsEighty patients in the Thoracic Surgery Department of First Hospital of Lanzhou University from December 2017 to February 2019 who were diagnosed lung cancer were divided into two groups. Forty patients in the MDT group were treated with MDT discussion. The control group consisted of 40 patients who were treated without MDT discussion. The incidence of postoperative complications and clinical efficacy were compared between the two groups . ResultsThere was no statistical difference in incision infection, atelectasis, pleural effusion and pulmonary leakage between the two groups. However, the incidence of postoperative pulmonary infection (5% vs. 20%, P=0.043) and the overall postoperative complications (17.5% vs. 42.5%, P=0.015) in the MDT group was lower than that in the control group with a statistical difference. In the MDT group, the operative time (140.3±8.0 min vs. 148.8±6.8 min, P<0.001), intraoperative bleeding ( 207.8±19.4 mL vs. 222.0±28.3 mL, P=0.010), lymph node dissection number (25.1±6.2 vs. 20.1±7.0, P=0.001), postoperative drainage (273.0±33.5 mL vs. 24.0±52.5 mL, P<0.001), drainage duration (81.9±6.1 h vs. 85.3±8.1 h, P=0.039), pain on the first day after surgery (4.6±0.7 vs. 5.4±0.7), P<0.001), pain on the second day (2.5±0.7 vs. 3.0±0.8, P=0.002), pain on the third day (1.1±0.8 vs. 1.5±0.6, P=0.014), postoperative activity time (40.7±6.7 h vs. 35.3±7.1 h, P<0.001), postoperative recovery time (6.8±0.9 d vs. 7.4±0.7 d, P=0.003), patient satisfaction (8.1±1.4 vs. 7.2±2.0, P=0.020) were significantly better than those of the control group with statistical differences. But there was no statistical difference in the conversion to thoracotomy between the two group. ConclusionMDT discussion can reduce the surgical risk and postoperative complications, improve the clinical efficacy and accelerate the postoperative rehabilitation of patients, which has a good clinical significance.
ObjectiveTo investigate the effect of Wnt5a derived from tumor-associated fibroblasts （CAFs） on the migration and invasion of gastric cancer cells. MethodsThe differentially expressed genes Wnt5a between CAFs and normal gastric fibroblasts (NGFs) in gastric cancer tissues and their corresponding normal gastric tissues using the GEO database GSE194261 dataset were screened. Immunohistochemical method was used to detect the expression of Wnt5a protein in tissue samples of clinical gastric cancer patients, and the relationship between Wnt5a protein expression and clinicopathological features of gastric cancer was analyzed. CAFs and NGFs were extracted from fresh surgical specimens of gastric cancer patients, and the expression of Wnt5a in CAFs was detected by real-time fluorescence quantitative-polymerase chain reaction and Western blot experiment. Transwell invasion and migration experiment was used to observe the effects of CAFs, inhibition of Wnt5a expression in CAFs and different concentrations of recombinant Wnt5a protein on the migration and invasion ability of gastric cancer MGC-803 and MKN-28 cell lines in vitro. ResultsThrough the screening of GEO database GSE194261 data set, it was found that Wnt5a was more expressed in CAFs than NGFs (P<0.05). Immunohistochemical results showed that the expression of Wnt5a protein in gastric cancer tissues was significantly stronger than that in normal gastric tissues (P<0.05), and the expression of Wnt5a protein was related to T stage of tumor (χ2=5.035, P<0.05), but not related to gender, age, degree of tumor differentiation, lymph node metastasis, vascular invasion and nerve invasion (P>0.05). Inhibiting Wnt5a derived from CAFs could inhibit the invasion and migration of gastric cancer cells. By stimulating gastric cancer cells with different concentrations of human recombinant Wnt5a protein, it was found that when the concentration of human recombinant Wnt5a protein was greater than 100 ng/mL, the invasion and migration abilities of MGC-803 and MKN-28 gastric cancer cells were significantly increased (P<0.05). ConclusionWnt5a is highly expressed in CAFs derived from the interstitial tissue of gastric cancer, which is related to the invasion depth of gastric cancer and can promote the invasion and migration of gastric cancer cells.
【Abstract】ObjectiveTo establish the stable model of orthtopic liver transplantation in rats.MethodsIn the light of Kamada’s method, the donor’s liver was perfused through portal vein before it was harvested,and the anastomosis was modified as continous suture with one suture for the suprahepatic inferior vena cava. Two hundred and ten orthotopic liver transplantations were performed in Wistar male rats according to this method. ResultsThe mean time of donor operation was 35 min, and that of recipient operation was 51 min. The mean cold preserving time of graft was 60 min.The anhepatic phase was about 17 min 〔(17.6±4.5) min〕. Nineteen rats died during operation. The causes of death included: bleeding of suprahepatic inferior vena cava, infrahepatic vena cava thrombosis, biliary obstruction, portal vein constriction and thrombosis, liver injury, bleeding of left subphrenic vein, infection, excessively deep anesthesia and respiratory failure. The 24hour survival was 91.0%(191/210),and the oneweek survival was 85.2%（179/210）.ConclusionThrough the modification of the anastomosis of suprahepatic inferior vena cava, the nonhepatic time of the recipients could be shortened,and the complications could be decreased. In this way, the survival of recipient rats after liver transplantation could be increased.
Objective To explore the influence of different withdrawal time of trachea cannula on percutaneous dilational tracheostomy (PDT) in critically ill patients. Methods In this study, we retrospectively analyzed the clinical data of 185 critically ill patients experienced PDT, who had been admitted to the adult mixed ICU of Xiaolan Hospital of Southern Medical University from January 2015 to July 2017. The patients were divided into an early PDT group (EPDT group) and a delayed PDT group (DPDT group) according to the timing of withdrawing trachea cannula. Operation information such as operation time, blood loss and the incidence of complications were collected and compared between the two groups. Results Between the EPDT group and the DPDT group, there were no obvious differences in operation time (minutes: 6.5±2.6 vs. 7.3±3.5), amount of blood loss (ml: 5.2±2.8 vs. 6.0±3.4) or conversion to traditional operation (1.9% vs. 2.4%) (all P>0.05). Compared with the EPDT group, the DPDT group patients experienced more fluctuation of intraoperative vital signs, used more dose of sedative and analgesic drugs, and experienced higher occurrence of aspiration (18.3%vs. 5.6%), balloon burst (13.4% vs. 2.9%), guide-wire placing difficulty (11.0% vs. 1.9%) and tracheostomy cannula placing difficulty (14.6% vs. 2.9%) (all P<0.05). There were no statistical significances in postoperative complications such as postoperative-hemorrhage, pneumothorax, pneumoderm, the posterior tracheal injury or incision infection between the two groups (allP<0.05). More patients acquired postoperative pulmonary infection in the DPDT group than the EDPD group (12.2%vs. 5.8%, P>0.05), and there was no statistical significances in mechanical ventilation time between the two groups (days: 5.5±3.0vs. 6.0±2.5, P>0.05). Conclusions The operation and complications of PDT in critically ill patients are influenced by the timing of withdrawing trachea cannula. The standard procedure of withdrawing trachea cannula preoperatively may offer better clinical operability and lower technical risk.
ObjectiveTo investigate the clinical effect of metal supported multi-sided versus ordinary ultra-fine drainage tube in the uniportal video-assisted thoracic surgery (VATS) lower pulmonary lobectomy. MethodsFrom January 2021 to June 2022, the clinical data of patients who underwent uniportal VATS lower lobectomy in our hospital were retrospectively analyzed. According to the different types of ultra-fine drainage tubes used in the surgery, the patients were divided into an experimental group (using multi-sided hole 10F ultra-fine drainage tubes with metal support) and a control group (using ordinary 12F ultra-fine drainage tubes). The clinical data of the two groups were compared. ResultsA total of 190 patients were enrolled, including 108 males and 82 females. There were 90 patients in the experimental group aged 56.60±10.14 years; and 100 patients in the control group aged 57.07±11.04 years. The incidences of postoperative lung infection and pleural effusion in the experimental group were lower than those in the control group, with statistically significant differences (P<0.05). The postoperative visual analogue scale score, the need to adjust the chest drainage tube after the surgery, the need for chest puncture after the surgery, the time of postoperative chest tube removal, and the hospitalization cost were statistically different (P<0.05). There was no statistical difference in the length of postoperative hospital stay or the incidences of postoperative lung leakage, arrhythmia, and atelectasis complications (P>0.05). ConclusionCompared with the ordinary ultra-fine drainage tubes, multi-sided hole ultra-fine drainage tubes with metal support can reduce the incidences of lung infection and pleural effusion complications after the uniportal VATS lower lobectomy, reduce the pain and economic burden, which can be applied in the uniportal VATS lower lobectomy.