A new and effective method to produce transgenic animals was established. Without a surgical incision, the recombinant plasmid containing green fluorescence protein (GFP) cDNA was repeatedly injected into male mouse testis at multi-sites. After few weeks of the final injection, the injected male was mated with normal oestrus female to produce transgenic mice. The presence of the GFP cDNA in F1 transgenic individuals were detected by polymerase chain reaction and Southern blot hybridization, which showed that the transgenic rate of mouse F1 offspring was 41%. The transferred gene was integrated into the host genome and could be transmitted to its offspring. When the positive F1 individuals were mated with the wild type ICR mice, the F2 individuals had a transgenic rate of 37%. The results indicate that the high efficiency of gene transfer and the limited number of manipulations make the method suitable for creating a large number of transgenic animals, especially, for producing domestic animals.

Objective To observe the morphological changes of pancreatic tissue of thoracic duct ligated rats in fine and ultrastructural levels, and to determine whether lymph block animal model can affect pancreatic islet amyloid polypeptide(PIAP)deposit in rat pancreas. Methods At the 6th month after the operation, some pancreatic tissue sections of 16-month-old experimental rats were embedded in paraffin wax and stained with HE and Congo red;immunohistochemical staining was performed on some frozen sections, which were then observed with light microscope;transmission electron microscope (TEM) specimen preparation and observation were performed on other samples. Results HE and Congo red stained sections showed that the pancreatic glandular lobule space was widened, with significant connective tissue hyperplasia, and fat accumulation when the islet was stained indistinctly or vermeil and tissue space was broadened. The sections with immunohistochemical staining displayed the pancreatic islet as well as the tissues around it were stained into dark brown being positive with PIAP antigen. TEM observation showed the pancreatic glandular interlobule space was widened, while blood vessels and enlarged lymphatic vessels were visible;within widened pancreatic islet interstitial space, a great quantity of lipid droplets and some collagen fibril structures could be seen.Conclusion The ligation of thoracic duct can contribute to pancreatic lymph draining block, lymphagiectasis, connective tissue space and interstitial space widening, fat accumulation, and PIAP deposit in rat pancreas. These structural changes may affect the function of pancreatic islets.

Objective To analyze the change in the concentration of IL-17 and IL-10 inflammatory factors among the deadaptation personnel who returned from the plateau.Methods A total 21 healthy males were investigated who averaged 25 years in age, lived permanently in the plains (200 m), and once stayed to the plateau (Lasha) for 6 months.Their venous blood was collected at three time points:the day before ascending to the plateau(control), the second day after return to the plains(d2) and the 30th day(d30), respectively.Their serum was seperated from the whole blood and the level of IL-17A and IL-10 was detected by ELISA method.Results The concentration of IL-17A and the IL-17A/IL-10 ratio were significant increased at d2 and d30, respectively, compared with control (P<0.05).Compared with d2, IL-17A and the IL-17A/IL-10 ratio were decreased obviously at d30(P <0.05).The level of IL-10 at d2 and d30 was significantly reduced compared with control ( P <0.05), but increased at d30 compared with d2.Correlative analysis showed that there was a negative correlation in the levels of IL-10 and IL-17A between control, d2 and d30, respectively (r1=0.948, P<0.05;r2=0.969, P<0.05;r3=0.972, P<0.05).A significant negative correlation was observed in the alteration levels of IL-10 and IL-17A between the three groups(r4=-0.793, P<0.05; r5=-0.756, P<0.05). Conclusion The concentration of inflammatory factors among the plateau deadaptation patients is imbalanced, but it is gradually reduced with time.The mechanism is still not clear.

Objective To evaluate the effect and mechanism of bone morrow MSCs transplantation in swine with AMI by cell biology and molecular imaging methods including PET/CT, SPECT, and MRI. Methods Twenty?four Chinese mini?swine ( ( 25 ± 5 ) kg ) were randomly divided into 2 groups: MSCs group ( n=12) and control group ( n=12) . Myocardial infarction was induced in swine hearts by occlusion of the LAD. Thirty minutes later, the MSCs group received autologous MSCs transplantation through in?tramyocardial injection into the peri?infarcted areas (2×107,2 ml) and the control group was subjected to cell culture medium in the same way. At the 1st and 4th weeks after MSCs transplantation, myocardial glu?cose metabolism, myocardial perfusion and cardiac function were evaluated in the two groups through PET/CT, SPECT and MRI. The minimum FDG mean signal intensity ( MSI ) , summed MSI, SRS, SRS%, LVEF, ESV, stroke volume ( SV) and cardiac output ( CO) were calculated. On the 4th week, HE and Masson′s Trichrome stains were performed. Mann?Whitney u test and non?parametric Wilcoxon test were used. Results (1) As evaluated by PET in the 1st week, the MSI and summed MSI in MSCs group were less than those in control group ( 22. 10 ± 3. 18 vs 35. 70 ± 3. 02, z=-2. 65; 1 013. 50 ± 29. 37 vs 1 084. 00 ± 21?15, z=-1.97;both P<0.05) . Compared to the minimum MSI and summed MSI in the 1st week, those in MSCs group increased significantly (34.00±4.25, z=-2.81;1 075.50±28.30, z=-2.80;both P<0?01) in the 4th week. SRS and SRS% decreased in the 4th week compared to those in the 1st week (20.20±2.24 vs 23.80±1.58, (29.80±3.31)% vs (35.10±2.34)%;both z=-2.08, both P<0.05). The averaged MSI in left ventricular infarction area (MSI<70) also increased (56.25±3.54 vs 48.14±2.71;z=-2.80, P<0.01). The a?bove?mentioned parameters had no statistically significant differences in the 4th week compared to those in the 1st week in the control group (all P>0.05). (2) In the 1st week, the perfusion variables had no signifi?cant differences between the two groups ( P>0.05) . There was no significant difference in any perfusion vari?ables between the 1st and 4th weeks in the two groups, respectively (P>0.05). (3) As evaluated by MRI, the cardiac functional parameters had no significant differences between the two groups at the 1st week. In the MSCs groups, LVEF increased significantly ((54.41±2.62)% vs (47.54±2.43)%;z=-2.60, P<0.01) and ESV reduced significantly ((22.85±1.91) vs (27.07±1.67) ml;z=-2.70, P<0.01) in the 4th week com?pared to those in the 1st week; SV and cardiac CO in the 4th week also increased significantly ((29.35± 1?84) vs (26.52±1.46) ml, (2.23±0.14) vs (1.96±0.13) L/min;z=-2.09 and -1.99, both P<0?05). In the control group, there were no significant differences in the cardiac functional parameters between the 1st and 4th weeks ( all P>0.05) . Conclusions Four weeks after MSCs transplantation for AMI, cardiac func?tion and myocardial glucose metabolism improved significantly but without significant myocardial perfusion improvement. Therefore, the cardiac function improvement might be associated with increased myocardial glucose metabolism.

Objective　To observe killing effect of HSV1-TK／GCV system on human pulmonary adenocarcinoma cell A549 in vitro and in vivo. Methods　A retroviral vector containing TK gene was constructed and transduced into pulmonary adenocarcinoma cell A549 by electroporation. The sensitivity of transfected cell to GCV in vitro and bystander effect and cellular apoptosis were observed. The recombination and expression of TK gene were examined by DNA PCR and in situ hybridization individually. In addition, the therapeutic effect of GCV on subcutaneous tumors inoculated with transfected and parental cells respectively was observed. Results　The transfected cells were irregular in shape, polyangular and easy of vacuolization. The double time of A549,A549-PLXSN and A549-TK was 36.15±3.27、40.82±3.75 and 42.06±4.12 hours respectively (P＞0.05). The sensitivity of transfected cells to GCV was 46 times higher than that of parental cells and bystander effect was more apparent in high density inoculation cells than in low density. Apoptotic bodies and semimoon feature in nuclear were observed in transfected cells, but not in parental cell. Apoptotic cells were found significantly more in transfected cells than in parental cells by FCM and TUNEL (P＜0.001). The recombination and expression of TK gene were positive in the transfected cells. In vivo, growth of tumors which formed by transfected cells was significantly inhibited by GCV, however, there was no similar inhibitive effect found in control group. Conclusion　The transfected cells have obtained sensitivity to GCV. The killing effect of TK／GCV system on tumor cells is probably related to apoptosis. GCV could inhibit growth of tumors inoculated by transfected cells.

[Abstract] Objective： ： To explore miR-103a-3p expression in the tumor tissues and the serum of breast cancer patients, and its role and mechanism in breast cancer development. Methods: Pathologically confirmed 31 cases of tumor tissues and 21 cases of para-cancerous tissues resected at Department of Oncological Surgery of the Second Affiliated Hospital of Hainan Medical University (Haikou, China) from March 1, 2017 to August 31，2017 were collected for this study; in addition, serum samples from 38 breast cancer patients and 22 healthy subjects as well as the breast cancer cell lines MCF-7 and MDA-MB-231 were used in this study. pHBLV-U6-Luc-T2A-Puro and PLL3.7 lentivirus were applied to knock down miR-103a-3p and PDK4 in MCF-7 and MDA-MB-231 cells, respectively. qPCR and Western blotting were performed to examine the mRNA and protein expressions of miR-103a-3p and PDK4 in tissues and serums of breast cancer patients as well as the in cell lines, respectively; CCK-8 assay was applied to detect the proliferation of MCF-7 and MDAMB-231 cells; Olympus AU5400 was applied to detect the glucose consumption and lactate production in indicated cell line. Results： ： miR-103a-3p was significantly decreased in tumor tissues compared with the paracancerous tissues (P<0.01). miR-103a-3p knockdown activated the glucos consumption and lactate production (all P<0.01), increased the PKD4 expression (P<0.01) in MCF-7 and MDAMD-231 cells, and promoted the proliferation of MCF-7 and MDA-MB-231 cells (P<0.01). Furthermore, knockdown of PDK4 suppressed the glucose consumption, lactate production and proliferation in MCF-7 and MDA-MB-231 cells with miR-103a-3p silencing (all P<0.01). Conclusion： ：In the breast cancer, miR-103a-3p inhibited the proliferation of breast cancer cells through down-regulation of PDK4 and PDK4-mediated aerobic glycolysis.

Objective:Explore the feasibility and advantages of using proximal and distal ends of thoracodosal artery and vein as recipient vessels in deep inferior epigastric perforator flap immediate breast reconstruction.Methods:The clinical data of patients who underwent breast reconstruction surgery using the proximal and distal ends of the thoracodorsal vein as recipient vessels at the Department of Breast and Thyroid Surgery of the Second Affiliated Hospital of Hainan Medical University from March 2022 to June 2023 were analyzed retrospectively. Preoperative examinations included thoracoabdominal angiography and color Doppler ultrasonic localization of the main trunk and perforators of the inferior epigastric vessels. The procedure began with mastectomy and axillary lymph node dissection, followed by the isolation of bilateral perforators and the main trunk of the abdominal flap. The main trunks of the bilateral inferior epigastric arteries were then transected, and their vascular pedicles exposed and anastomosed respectively to the proximal and distal ends of the thoracodorsal artery and vein. Both arteries and veins were joined end-to-end. The flap after trimming and reconstruction was then implanted into the cavity left after mastectomy through the incision. Breast positioning was performed with the patient in a knee-bent and hip-flexed position. After adjusting the shape of the reconstructed breast. The donor site was closed, the umbilicus was reconstructed, drainage tubes were placed, and the breast incision was closed. Postoperative follow-up monitored complications associated with the flap and patient satisfaction with the breast reconstruction, utilizing a self-assessment method.Results:Four female patients were included, aged (46.0±6.5) years, ranging from 37 to 52 years. All four patients had bilateral vascular pedicles in the donor area, with three patients having thoracodorsal vessels at the distal and proximal ends as recipient vessels, and one patient having anterior serratus branch of the thoracodorsal vessels at the distal and proximal ends. All drainage tubes were removed within 7 to 10 days after surgery. Patients were discharged. Follow-up period ranged from 1 to 15 months, averaging 6 months. The patients recovered well postoperatively, with no flap-related complications occurring. All four patients were satisfied with the result of the reconstruction.Conclusion:The simultaneous application of the proximal and distal ends of the thoracodorsal artery and vein can ensure the safety of flap survival while reducing damage to the ribs and intercostal muscles, achieving better aesthetic result.

Objective:Explore the feasibility and advantages of using proximal and distal ends of thoracodosal artery and vein as recipient vessels in deep inferior epigastric perforator flap immediate breast reconstruction.Methods:The clinical data of patients who underwent breast reconstruction surgery using the proximal and distal ends of the thoracodorsal vein as recipient vessels at the Department of Breast and Thyroid Surgery of the Second Affiliated Hospital of Hainan Medical University from March 2022 to June 2023 were analyzed retrospectively. Preoperative examinations included thoracoabdominal angiography and color Doppler ultrasonic localization of the main trunk and perforators of the inferior epigastric vessels. The procedure began with mastectomy and axillary lymph node dissection, followed by the isolation of bilateral perforators and the main trunk of the abdominal flap. The main trunks of the bilateral inferior epigastric arteries were then transected, and their vascular pedicles exposed and anastomosed respectively to the proximal and distal ends of the thoracodorsal artery and vein. Both arteries and veins were joined end-to-end. The flap after trimming and reconstruction was then implanted into the cavity left after mastectomy through the incision. Breast positioning was performed with the patient in a knee-bent and hip-flexed position. After adjusting the shape of the reconstructed breast. The donor site was closed, the umbilicus was reconstructed, drainage tubes were placed, and the breast incision was closed. Postoperative follow-up monitored complications associated with the flap and patient satisfaction with the breast reconstruction, utilizing a self-assessment method.Results:Four female patients were included, aged (46.0±6.5) years, ranging from 37 to 52 years. All four patients had bilateral vascular pedicles in the donor area, with three patients having thoracodorsal vessels at the distal and proximal ends as recipient vessels, and one patient having anterior serratus branch of the thoracodorsal vessels at the distal and proximal ends. All drainage tubes were removed within 7 to 10 days after surgery. Patients were discharged. Follow-up period ranged from 1 to 15 months, averaging 6 months. The patients recovered well postoperatively, with no flap-related complications occurring. All four patients were satisfied with the result of the reconstruction.Conclusion:The simultaneous application of the proximal and distal ends of the thoracodorsal artery and vein can ensure the safety of flap survival while reducing damage to the ribs and intercostal muscles, achieving better aesthetic result.

Objective To investigate the characteristics of liver deformation and changes in abdominal vascular diameter under artificial pneumoperitoneum based on CT images,and also to reveal the impact of intestinal adhesion on these characteristics.Methods A retrospective study was conducted on 61 patients who underwent abdominal CT scan and artificial pneumoperitoneum CT.Eight cases did not meet the inclusion criteria.The sample included 41 patients with postoperative intestinal adhesion(POIA)and 12 patients without POIA.A three-dimensional post-processing software was used to quantify the gas volume,liver volume,liver surface area,and lumen of the abdominal aorta,inferior vena cava,and portal vein on CT images of patients who underwent abdominal CT scan and artificial pneumoperitoneum CT.The measured indices were compared before and after artificial pneumoperitoneum in all patients,and the impact of intestinal adhesion on the aforementioned indices was evaluated.Results At equivalent levels of pneumoperitoneum pressure,no statistically significant difference was observed in the quantity of gas in the abdominal cavity between patients with and without POIA(P=0.753).In the absence of pneumoperitoneum,POIA patients exhibited a significantly higher liver volume than non-adhesion patients(P＜0.01).Following the application of artificial pneumoperitoneum pressure,both liver volume and liver surface area were reduced in all patients,with a more pronounced decrease in POIA group(P=0.003,P=0.002).In addition,the reductions in the internal diameters of the inferior vena cava at three locations and the portal vein position 1 were also observed in all patients(P＜0.05).Notably,the POIA group exhibited more significant decrease in the internal diameters of inferior vena cava position 1 and the portal vein position 1(P=0.022,P=0.038).No significant differences were observed in the changes of internal diameter of other blood vessels or in the intergroup comparison.Conclusion Artificial pneumoperitoneum can reduce the volum,the surface area of liver,and the inner diameter of some abdominal vessels,which provides a scientific basis for navigation and real-time correction in laparoscopic abdominal surgery for patients,especially for patients with POIA.