ObjectiveTo evaluate the most efficient method for transfection of human umbilical cord mesenchymal stem cells (HUMCSs) in vivo. MethodsHUCMSCs were isolated from human umbilical cord and cultured, which were labelled by PKH26 and lentivirus-GFP, then were observed by using a fluorescence microscope. Sixty SD rats were randomly divided into PKH26 transfection group and lentivirus-GFP transfection group. The right hepatic lobe of rat was resected, then the transfected stem cells were injected into portal vein. The rats were sacrificed on day 3, 8, and 13 after transfection. The liver specimens were observed by using a fluorescence microscope. Flow cytometry was used to evaluate the percentage of transfected stem cells and the apoptotic stem cells. ResultsThe third generation of HUCMSCs labelled by PKH26 and lentivirus-GFP were spindle shaped. PKH26 red dye was evenly distributed in the cell membrane of HUCMSCs and could be clearly labelled. The HUCMSCs labelled by lentivirus-GFP were green fluorescence under the fluorescence microscope, and it was clear and stable. The HUCMSCs were clear and could be clearly distinguished on day 3 after transfection by two methods in vivo. As the time went by, red was faded and blurred, then was gradually disappeared on day 13 after transfection in the HUCMSCs stansfected by PKH26; but the color in the HUCMSCs stansfected by lentivirus-GFP were clear at all the time points. The transfection rate of the lentivirus-GFP was significantly higher that that of the PKH26 (P < 0.05), the rate of apoptotic stem cells had no significant differences at all the time points between these two groups (P > 0.05). ConclusionLentivirus-GFP transfection is a higher efficient method for stem cell labelling in vivo, it could be used to observe transplantation cells for a long time in future.
Objective To explore the safety and feasibility of using modified Glisson pedicle transection methodin the precise hepatectomy of hepatocellular carcinoma （HCC）. Methods Sixty patients with HCC， which confirmed by postoperational pathology were admitted in the study. During the surgery of experimental group （Glisson group）， the segment pedicle were transected firstly using modified Glisson pedicle transection method. Then， the liver parenchyma was split follow the hepatic vein guided by intraoperative ultrasound. During the surgery of contrast group （Prigle group），the liver parenchyma was split using ultracision harmonic scalpel under intermittent pringle clamping of hilar. Results There were no significant difference in the amount of intraoperative bleeding， blood transfusion， as well as duration of surgery， serum alanine aminotransferase， total bilirubin， and length of hospital stay between the two groups （P＞0.05）. However， the incidence of postoperative complication was lower in Glisson group （23.3% vs. 50.0%， P＜0.05）. In addition， the length of tumor margin was more favorable in Glisson group 〔（2.3±0.7） cm vs. （1.5±0.6） cm， P＜0.05〕. The recurrence rate of Glisson group was lower than that Prigle group， but was not different significantly （P＞0.05）. Conclusions The modified Glisson pedicle transection method has the same safety as traditional method in the precise hepatectomy of HCC. And it has the advantages in lower postoperative complication and more favorable tumor margin， which may reduce the recurrence rate theoretically.
Objective To explore the value of pathologic diagnosis for pancreatic head mass by using recise Tru-cut biopsy under intraoperative ultrasound guided. Methods Twenty-eight patients with solid pancreatic masses in People’s Hospital of Suqian，Affiliated Hospital Xuzhou Medical College from August 2010 to August 2011 were performed precise Tru-cut biopsy under intraoperative ultrasound guided. In all patients of 28 cases, male 20 cases and female 8 cases, the male-to-female ratio was 5∶2. The patients’ age was 34-78 years old（mean age： 64 years old）. Twenty-eight patients were divided into three groups based upon the greatest dimension of the masses as follows：equal or less than 1.5 cm （group S, 5 cases）, 1.5-3.0cm （group M, 7 cases）, and greater than 3.0cm （group L, 16 cases）. Three needlepasses in each mass were performed. The results of postoperative pathologic findings were compared with specimens in paraffin sections. Results In all cases of 28, the diagnostic coincidence rate was 100%, there were no false positive finding and false negative. The coincidence ratio for pathological diagnosis of tissues with only 1 strip sample （1/3）, only 2 strip samples （2/3）, and with 3 strip samples （3/3） were 3/5, 2/5, and 0/0, respectivly in S group, 0/0, 5/7, and 2/7, respectivly in M group, and 0/0, 4/16, and 12/16, respectivly in L group. The false negative rate of single strip sample in S group and M group was higher than that in L group （χ2=9.833，P=0.002）. There was false negative finding with master single test in small focus of infection. Conclusion Precise Tru-cut biopsy under intraoperative ultrasound guided is a safe and highly accurate method for pathological diagnosis of patients with solid pancreatic lesions, especially in small lesions，it is worthy of clinical application..
Objective To explore the effectiveness of limited small incision with simple Krackow suture in treatment of acute closed Achilles tendon rupture. Methods Between October 2013 and July 2016, 25 cases with acute Achilles tendon rupture were repaired by simple Krackow suture via limited small incision. There were 21 males and 4 females with an average age of 33.6 years (range, 25-39 years). The left side was involved in 15 cases and the right side in 10 cases. The injury caused by sport in 22 cases and by falling in 3 cases. The time from injury to operation was 3-7 days (mean, 4.4 days). Physical examination showed that the Thompson sign and single heel raising test were positive. Results The operation time was 30-60 minutes with an average of 39.2 minutes. All incisions healed by first intention. There was no complication of wound infection, deep vein thrombosis, tendon re-rupture, and sural nerve injury. All patients were followed up 9-20 months (mean, 14.2 months). The ankle and hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS) was 92-97 (mean, 94.9) after 9 months. The AOFAS score results were excellent in 13 cases, good in 9 cases, and fair in 3 cases. The range of motion of ankle joint was 49-58° with an average of 53.7°. All single heel raising tests were negative. Conclusion The method of simple Krackow suture via limited small incision has the advantages of minimal injury, less incidence of re-rupture and sural nerve injury, quicker recovery and so on.