Implementation of sodium alginate-Fe3O4 to localize undiagnosed small pulmonary nodules for surgical management in a preclinical rabbit model.

Many methods are used to locate preoperative small pulmonary nodules. However, deficiencies of complications and success rates exist. We introduce a novel magnetic gel for small pulmonary nodules localization in rabbit model, and furtherly evaluate its safety and feasibility. Rabbits were used as the experimental objects. A magnetic gel was used as a tracer magnet, mixed as sodium alginate-Fe3O4 magnetic fluid and calcium gluconate solution. In short-term localization, a coaxial double-cavity puncture needle was applied to inject the gel into the lung after thoracotomy, and a pursuit magnet made of Nd-Fe-B permanent magnetic materials was used to attract the gel representing location of the nodule. In long-term localization, the gel was injected under X-ray guidance. Imaging changes to the lung were observed under X-ray daily. Thoracotomy was performed to excise tissue containing the gel, and hematoxylin-eosin staining was used to observe the tissue on postoperative days 1, 3, 5, and 7. Observe tissues morphology of heart, liver, spleen, and kidney in the same way. The gel was formed after injection and drew lung tissue to form a protrusion from the lung surface under the applied magnetic field. No complication was observed. The shape and position of the gel had not changed when viewed under X-ray. Pathological analysis showed the gel had a clear boundary without diffusion of magnetic fluid. All tissues retained good histologic morphology and no magnetic fluid was observed. Our study preliminarily suggested that the technique using sodium alginate-Fe3O4 magnetic gel to locate small pulmonary nodules with guidance of X-ray, and to search for them under an applied magnetic field during the operation is safe and feasible.

Magnetic-assisted laparoscopic liver transplantation in swine.

BACKGROUND: Although laparoscopic technology has achieved rapid development in the surgical field, it has not been applied to liver transplantation, primarily because of difficulties associated with laparoscopic vascular anastomosis. In this study, we introduced a new magnetic-assisted vascular anastomosis technique and explored its application in laparoscopic liver transplantation in pigs. METHODS: Two sets of magnetic vascular anastomosis rings (MVARs) with different diameters were developed. One set was used for anastomosis of the suprahepatic vena cava (SHVC) and the other set was used for anastomosis of the infrahepatic vena cava (IHVC) and portal vein (PV). Six laparoscopic orthotopic liver transplantations were performed in pigs. Donor liver was obtained via open surgery. Hepatectomy was performed in the recipients through laparoscopic surgery. Anastomosis of the SHVC was performed using hand-assisted magnetic anastomosis, and the anastomosis of the IHVC and PV was performed by magnetic anastomosis with or without hand assistance. RESULTS: Liver transplants were successfully performed in five of the six cases. Postoperative ultrasonographic examination showed that the portal inflow was smooth. However, PV bending and blood flow obstruction occurred in one case because the MVARs were attached to each other. The durations of loading of MVAR in the laparoscope group and manual assistance group for IHVC and PV were 13 ± 5 vs. 5 ± 1 min (P < 0.01) and 10 ± 2 vs. 4 ± 1 min (P < 0.05), respectively. The durations of MVAR anastomosis in the laparoscope group and manual assistance group for IHVC and PV were 5 ± 1 vs. 1 ± 1 min (P < 0.01), and 5 ± 1 vs. 1 ± 1 min (P < 0.01), respectively. The anhepatic phase was 43 ± 4 min in the laparoscope group and 23 ± 2 min in the manual assistance group (P < 0.01). CONCLUSIONS: Our study showed that magnetic-assisted laparoscopic liver transplantation can be successfully carried out in pigs.

Plasma asprosin, CCDC80 and ANGPTL4 levels are associated with metabolic and cardiovascular risk in patients with inflammatory bowel disease.

Asprosin, coiled-coil domain-containing 80(CCDC80) and angiopoietin-like4(ANGPTL4) are newly discovered adipocytokine that affects glucose tolerance, insulin resistance and cardiovascular diseases. The goal of this study was to investigate if a relationship exists among asprosin, CCDC80 and ANGPTL4 and inflammatory bowel disease (IBD). Fifty subjects with newly diagnosed IBD and fifty healthy individuals were enrolled. Patients were treated with standard therapies for 3 months. Plasma asprosin, CCDC80 and ANGPTL4 levels were measured with enzyme-linked immunosorbent assay. High resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperemia (flow-mediated dilation, FMD) and after sublingual glyceryltrinitrate.Compare with healthy individuals, plasma CCDC80,erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) levels and homeostasis modelassessment of insulin resistance (HOMA-IR) were significantly higher (p < 0.05, respectively), whereas plasma asprosin,ANGPTL4 levels and FMD were significantly lower inboth UC and CD patients(p <0.05). Plasma CCDC80 levels were significantly higher in patients with CD (p<0.05), while plasma asprosin and ANGPTL4 levels were lower (p<0.05) as compared with those in patients with UC. Standard therapies increased plasma asprosin, ANGPTL4 levels and FMD in both UC and CD (p<0.05),UC and CD patientswhile decreased plasma CCDC80, ESR, CRP levels and HOMA-IR (p<0.05). The changes in HOMA-IR and FMD were correlated with the changes in plasma asprosin, CCDC80 and ANGPTL4 levels over the study period (p<0.05). Plasma asprosin, CCDC80 and ANGPTL4 levels may be applied as a significant marker for early stage of insulin resistance and atherosclerosis in IBD, especially of CD.

Predictive and prognostic implications of 4E-BP1, Beclin-1, and LC3 for cetuximab treatment combined with chemotherapy in advanced colorectal cancer with wild-type KRAS: Analysis from real-world data.

BACKGROUND: Colorectal cancer (CRC) is one of the main causes of cancer-related deaths in China and around the world. Advanced CRC (ACRC) patients suffer from a low cure rate though treated with targeted therapies. The response rate is about 50% to chemotherapy and cetuximab, a monoclonal antibody targeting epidermal growth factor receptor (EGFR) and used for ACRC with wild-type KRAS. It is important to identify more predictors of cetuximab efficacy to further improve precise treatment. Autophagy, showing a key role in the cancer progression, is influenced by the EGFR pathway. Whether autophagy can predict cetuximab efficacy in ACRC is an interesting topic. AIM: To investigate the effect of autophagy on the efficacy of cetuximab in colon cancer cells and ACRC patients with wild-type KRAS. METHODS: ACRC patients treated with cetuximab plus chemotherapy, with detailed data and tumor tissue, at Sun Yat-sen University Cancer Center from January 1, 2005, to October 1, 2015, were studied. Expression of autophagy-related proteins [Beclin1, microtubule-associated protein 1A/B-light chain 3 (LC3), and 4E-binding protein 1 (4E-BP1)] was examined by Western blot in CRC cells and by immunohistochemistry in cancerous and normal tissues. The effect of autophagy on cetuximab-treated cancer cells was confirmed by MTT assay. The associations between Beclin1, LC3, and 4E-BP1 expression in tumor tissue and the efficacy of cetuximab-based therapy were analyzed. RESULTS: In CACO-2 cells exposed to cetuximab, LC3 and 4E-BP1 were upregulated, and P62 was downregulated. Autophagosome formation was observed, and autophagy increased the efficacy of cetuximab. In 68 ACRC patients, immunohistochemistry showed that Beclin1 levels were significantly correlated with those of LC3 (0.657, P < 0.001) and 4E-BP1 (0.211, P = 0.042) in ACRC tissues. LC3 was significantly overexpressed in tumor tissues compared to normal tissues (P < 0.001). In 45 patients with wild-type KRAS, the expression levels of these three proteins were not related to progression-free survival; however, the expression levels of Beclin1 (P = 0.010) and 4E-BP1 (P = 0.005), pathological grade (P = 0.002), and T stage (P = 0.004) were independent prognostic factors for overall survival (OS). CONCLUSION: The effect of cetuximab on colon cancer cells might be improved by autophagy. LC3 is overexpressed in tumor tissues, and Beclin1 and 4E-BP1 could be significant predictors of OS in ACRC patients treated with cetuximab.

Magnetic Anastomosis Rings to Create Portacaval Shunt in a Canine Model of Portal Hypertension.

PURPOSE: This study evaluated a novel magnetic compression technique (magnamosis) for creating a portacaval shunt in a canine model of portal hypertension, relative to traditional manual suture. METHODS: Portal hypertension was induced in 18 dogs by partial ligation of the portal vein (baseline). Six weeks later, extrahepatic portacaval shunt implantation was performed with either magnetic anastomosis rings, or traditional manual suture (n = 9, each). The two groups were compared for operative time, portal vein pressure, and serum biochemical indices. Twenty-four weeks post-implantation, the established anastomoses were evaluated by color Doppler imaging, venography, and gross and microscopic histological examinations. RESULTS: Anastomotic leakage did not occur in either group. The operative time to complete the anastomosis for magnamosis (4.12 ± 1.04 min) was significantly less than that needed for manual suture (24.47 ± 4.89 min, P < 0.01). The portal vein pressure in the magnamosis group was more stable than that in the manual suture group. The blood ammonia level at the end of the 24-week post-implantation observation period was significantly lower in the magnamosis group than in the manual suture group. Gross and microscopic histological examinations revealed that better smoothness and continuity of the vascular intima had been achieved via magnamosis than with manual suture. CONCLUSION: Magnamosis was superior to manual suture for the creation of a portacaval shunt in this canine model of portal hypertension.

Serum HMGB1 levels and its association with endothelial dysfunction in patients with polycystic ovary syndrome.

High-mobility group box 1 (HMGB1) is newly discovered protein, which play a crucial role in the pathogenesis of systemic inflammation. Recent studies showed that HMGB1 is one of the important pathophysiological mechanisms in the occurrence and development of atherosclerosis. The purpose of the present study was to investigate the relationship between serum HMGB1 levels and endothelial function in patients with polycystic ovary syndrome (PCOS). Eighty newly diagnosed patients with PCOS and eighty normal women of similar age were selected. Metformin treatment (1,500 mg/day) was initiated in all patients for a period of consecutive 3 months. Serum HMGB1 levels were measured by ELISA. High resolution ultrasound was used to measure the brachial artery diameter at rest, after reactive hyperemia (flow-mediated arterial dilation, FMD) and after sublingual glyceryltrinitrate. Serum HMGB1 levels in PCOS were 24.87+/-14.93 ng/ml, which were significantly higher than that in controls (8.82+/-3.55 ng/ml, p<0.01). After 3 months treatment, serum HMGB1 levels decreased significantly (p<0.05). By dividing the distribution of HMGB1 levels into quartiles, serum HMGB1 levels were increased gradually with the increase of testosterone levels (p<0.05), whereas the FMD levels decreased (p<0.05). Multiple stepwise linear regression analysis showed that FMD (estimated coefficient beta=-0.69, p=0.005), testosterone (beta=0.31, p=0.045), TBARS (beta=0.69, p=0.012) and hs-CRP levels (beta=0.68, p=0.001) were significantly associated with HMGB1. The absolute changes in HMGB1 showed a positive correlation with the changes in testosterone (p<0.05) and negative correlation with the changes in FMD (p<0.05) in patients with PCOS during the course of metformin therapy. Serum HMGB1 levels are correlated with endothelial dysfunction in patients with PCOS. Our study suggests that HMGB1 may contribute to the early stage of atherosclerosis in patients with PCOS.

Magnetic Anastomosis for Biliojejunostomy: First Prospective Clinical Trial.

BACKGROUND: Magnetic compression anastomosis (magnamosis, MCA) has been verified safe and effective by us and others in animal bilioenteric anastomosis (BEA). The objective of the present study was to introduce clinical application of magnetic compression bilioenteric anastomosis (MC-BEA) with a unique device in series of patients. METHODS: Patients with obstructive jaundice with an indication of BEA were prospectively enrolled from 2012 to 2015. After dissection of bile ducts, the mother ring and drainage tube were placed in the proximal bile duct and the purse-string suture was tightened over the drainage tube. The drainage tube was introduced into the jejunal lumen at the anastomotic site and used to guide the daughter ring to assemble with the mother ring. All the patients were routinely followed up for magnets discharge or any complications associated. RESULTS: Forty-one patients were included. Thirty-four (82.9%) patients had a malignant primary disease, while seven (17.1%) had benign disease. The median time for MC-BEA was 10.5 min (interquartile range [IQR] 8.3-13.0 min). No perioperative morbidity or mortality associated with MC-BEA was observed. The median time for a patent bilioenteric anastomosis formation was 19.0 days (IQR 14.5-23.0 days), and the magnets were discharged with a median postoperative duration of 35.0 days (IQR 28.0-43.0 days). With a median follow-up of 547.5 days (range 223-1042 days), no patients had biliary fistula, while two (4.9%) developed anastomotic stricture at 4 months and 14 months after surgery, and underwent reoperation for reconstruction of BEA. CONCLUSIONS: MCA is a safe, effective, and time-saving modality for biliojejunostomy.

Treatment of rectovaginal fistula by magnetic compression.

INTRODUCTION AND HYPOTHESIS: Rectovaginal fistula (RVF) is an abnormal epithelium-lined connection between the rectum and vagina. The primary effective treatment is surgical repair, but recurrence remains a challenge. Magnetic compression anastomosis (MCA), an alternative to suturing, has been developed to generate an anastomosis between various hollow viscera. We hypothesized that the MCA approach could be used to treat RVF. METHOD: We designed a novel MCA device for RVF treatment and evaluated the magnetic compression procedure in a RVF pig model in comparison with the traditional suturing procedure. Following satisfactory outcomes, we also applied the MCA procedure to a human patient with recurrent RVF. The MCA device was designed based on the anatomical characteristics of the pig vagina and previous literature. The pig RVF model were established surgically (n = 12), and compression and control groups were each treated. The data were analyzed by one-way analysis of variance. RESULTS: qqExcept in one animal in each group, the RVF site was smooth and healing was complete. Histological analysis confirmed complete healing of the RVF with high histological continuity to neighboring tissues. The compression procedure applied to our patient with RVF was successful. The patient recovered quickly without complications, and RVF did not recur during a 15-month follow-up. CONCLUSIONS: From this preliminary investigation, MCA using our novel device appears to be a safe, simple, and effective nonsurgical procedure for the treatment of RVF.

Choledochojejunostomy with an innovative magnetic compressive anastomosis: How to determine optimal pressure?

AIM: To investigate the optimal magnetic pressure and provide a theoretical basis for choledochojejunostomy magnetic compressive anastomosis (magnamosis). METHODS: Four groups of neodymium-iron-boron magnets with different magnetic pressures of 0.1, 0.2, 0.3 and 0.4 MPa were used to complete the choledochojejunostomy magnamosis. Twenty-six young mongrel dogs were randomly divided into five groups: four groups with different magnetic pressures and 1 group with a hand-suture anastomosis. Serum bilirubin levels were measured in all groups before and 1 wk, 2 wk, 3 wk, 1 mo and 3 mo after surgery. Daily abdominal X-ray fluoroscopy was carried out postoperatively to detect the path and the excretion of the magnet. The animals were euthanized at 1 or 3 mo after the operation, the burst pressure was detected in each anastomosis, and the gross appearance and histology were compared according to the observation. RESULTS: The surgical procedures were all successfully performed in animals. However, animals of group D (magnetic pressure of 0.4 MPa) all experienced complications with bile leakage (4/4), whereas half of animals in group A (magnetic pressure of 0.1 MPa) experienced complications (3/6), 1 animal in the manual group E developed anastomotic stenosis, and animals in group B and group C (magnetic pressure of 0.2 MPa and 0.3 MPa, respectively) all healed well without complications. These results also suggested that the time required to form the stoma was inversely proportional to the magnetic pressure; however, the burst pressure of group A was smaller than those of the other groups at 1 mo (187.5 ± 17.7 vs 290 ± 10/296.7 ± 5.7/287.5 ± 3.5, P < 0.05); the remaining groups did not differ significantly. A histologic examination demonstrated obvious differences between the magnamosis groups and the hand-sewn group. CONCLUSION: We proved that the optimal range for choledochojejunostomy magnamosis is 0.2 MPa to 0.3 MPa, which will help to improve the clinical application of this technique in the future.

Fast magnetic reconstruction of the portal vein with allogeneic blood vessels in canines.

BACKGROUND: The resection and reconstruction of large vessels, including the portal vein, are frequently needed in tumor resection. Warm ischemia before reconstruction might have deleterious effects on the function of some vital organs and therefore, how to reconstruct the vessels quickly after resection is extremely important. The present study was to introduce a new type of magnetic compression anastomosis (MCA) device to establish a quick non-suture anastomosis of the portal vein after resection in canines. METHODS: The new MCA device consists of a pair of titanium alloy and neodymium-ferrum-boron magnet (Ti-NdFeB) composite rings. The NdFeB magnetic ring as a core of the device was hermetically sealed inside the biomedical titanium alloy case. Twelve canines were divided into two groups: a MCA group in which the end-to-end anastomoses was made with a new device after resection in the portal vein and a traditional manual suture (TMS) group consisted of 6 canines. The anastomosis time, anastomotic patency and quality were investigated at week 24 postoperatively. RESULTS: The portal vein was reconstructed successfully in all of the animals and they all survived. The duration of portal vein anastomosis was significantly shorter in the MCA group than in the TMS group (8.16+/-1.25 vs 36.24+/-2.17 min, P<0.05). Portography and ultrasound showed that the blood flow was normal without angiostenosis or thrombosis in all of the canines. Hematoxylin-eosin staining and electron microscope scanning showed in contrast to the TMS group, MCA anastomotic intimal was much smoother with more regularly arranged endothelial cells at week 24 postoperatively. CONCLUSIONS: The Ti-NdFeB composite MCA device was applicable in reconstruction of large vessels after resection. This device was easy to use and the anastomosis was functionally better than the traditional sutured anastomosis.

Tumor size is not a criterion for resection during the management of giant hemangioma of the liver.

BACKGROUND: Liver hemangiomas are the most common benign liver tumors. The management of giant (≥5 cm) hemangioma of the liver remains controversial. The aim of this study was to assess the influence of tumor size on postoperative outcomes after hepatectomy in patients with giant hemangioma of the liver. METHODS: Patients who were subjected to resection because of giant liver hemangioma between December 2006 and July 2012 were grouped by largest tumor size: 5-10 cm (group A) and 10-15 cm (group B). All patients underwent detailed preoperative assessments. Clinicopathologic features were analyzed, and univariate and multivariate analyses were used to determine risk factors that correlated independently with any complication, as well as the intraoperative red blood cell transfusion requirement. Long-term outcomes were assessed with a median follow-up of 56 months. RESULTS: One hundred and ninety patients, mean age 46 years, were included. The 146 patients with tumors 5-10 cm in size were compared with the remaining 44 patients with tumors 5-10 cm in size. The differences in postoperative morbidity (29.86 vs. 41.30%, P=0.150) and duration of hospitalization (11.06±7.02 vs. 12.17±7.74, P=0.465) between group A and group B did not reach statistical significance. Operation time, blood loss, and transfusion volume of group B were greater than those of group A. No perioperative deaths occurred and no recurrences were registered during follow-up in both groups. The results of univariate and multivariate analysis showed that diameter was not an independent risk factor of postoperative complications and intraoperative red blood cell transfusion. CONCLUSION: Giant hemangiomas should be monitored regularly. Asymptomatic tumors 5-10 cm in diameter can be managed conservatively even though they grow. When necessary, surgical treatment can be well justified because of low morbidity and mortality.

Relationship between serum irisin levels and urinary albumin excretion in patients with type 2 diabetes.

AIM: Irisin is first discovered as a potential mediator of obesity related energy homeostasis. Recent studies indicate that irisin is associated with endothelial dysfunction and atherosclerosis in patients with type 2 diabetes. Our objective was to examine the relationship between irisin and urinary albumin excretion in patients with type 2 diabetes. METHODS: 100 newly diagnosed patients with type 2 diabetes and 100 healthy subjects were selected. Serum irisin levels were measured by ELISA, and urine albumin was measured by radioimmunoassay. High resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperemia (flow-mediated arterial dilation, FMD) and after sublingual glyceryltrinitrate. RESULTS: Patients with type 2 diabetes presented decreased irisin levels when compared to controls (14.12±3.93 versus 28.98±2.56ng/ml, P=0.015).Serum irisin levels in the microalbuminuric and macroalbuminuria subgroup were 9.89±1.56ng/ml and 5.67±1.89ng/ml, respectively, which were significantly lower than those in the normoalbuminuria (15.97±3.12ng/ml). In comparison to microalbuminuric subgroup, macroalbuminuria subgroup had lower levels of irisin. By dividing the distribution of serum irisin levels into quartiles, FMD was increased gradually with the increase of serum irisin levels (P<0.001). Multiple stepwise linear regression analysis showed that FMD (ß=0.75, P=0.002), 2-hBG (ß=-0.25, P=0.038) and UAE (ß=-0.87, P=0.008) were significantly associated with irisin. Pearson's correlation analyses showed a negative correlation between irisin and logUAE (r=-0.57) and between FMD and logUAE (r=-0.47), and positive correlations between irisin and FMD (r=0.51). CONCLUSIONS: Decreased plasma levels of irisin seem to be associated with UAE and FMD in patients with type 2 diabetes.

Changes of plasma concentration of osteoprotegerin and its association with endothelial dysfunction before and after hypouricemic therapy in patients with hyperuricemia.

OBJECTIVE: Osteoprotegerin (OPG) is a secreted glycoprotein in the regulation of bone turnover. Recently, many studies showed that OPG acts as an important regulatory molecule in the vascular systems. Our objective was to examine the plasma OPG levels alteration and its association with endothelial function before and after hypouricemic therapy in patients with hyperuricemia. METHODS: Thirty patients (28 males and 2 females, serum uric acid > 7.0 mg/dl) with hyperuricemia were selected. Thirty healthy individuals (28 males and 2 females) with normal serum uric acid were also selected as control. Patients were administered with hypouricemic therapy for 6 months. Plasma OPG concentration was measured in duplicate using a sandwich ELISA and high-resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperemia and after sublingual glyceryltrinitrate. RESULTS: Plasma OPG levels in patients with hyperuricemia before hypouricemic therapy was significantly higher than those in controls (3.39 ± 0.25 vs. 2.05 ± 0.74 ng/L, p < 0.01). After hypouricemic therapy, OPG levels decreased markedly (2.54 ± 0.38 ng/L, p < 0.01). Flow-mediated dilation (FMD) in patients with hyperuricemia was 3.07 ± 1. 23%, which was significantly lower than that in control subjects (4.62 ± 0.69%, p < 0.01), and it improved significantly after hypouricemic therapy (3.91 ± 1.37%, p < 0.01). The absolute changes in OPG showed a significant positive correlation with the changes in serum uric acid (p < 0.05) and negative correlation with the changes in FMD (p < 0.01) in patients with hyperuricemia during the course of hypouricemic therapy. CONCLUSION: The current study demonstrates that plasma OPG levels increased significantly in patients with hyperuricemia and decreased significantly after hypouricemic therapy, and are correlated with FMD. These findings support the growing concept that elevated plasma OPG levels may be involved with the development of endothelial dysfunction in patients with hyperuricemia.

[Screening and identification of CD13-binding peptides with phage display peptide library].

OBJECTIVE: To screen and identify the peptides that specifically bind to CD13 on monocytes. METHODS: The phages capable of specific binding to CD13 were screened in the phage-displayed 12-peptide library. The affinity of the selected phages with CD13 was verified with enzyme-linked immunosorbent assay (ELISA). The sequences of the peptides bound to the phages were deduced according to the phage DNA sequences, and the functional peptides aligned using the BLASTP on the Website NCBI were synthesized. To analyze the biological function of the screened peptides, the location of the peptides bound to THP-1 cells was detected using immunofluorescence assay. The blocking effect of WM15 on the peptide binding to THP-1 cells was assessed by immunofluorescence assay. RESULTS: The phages that specifically bound to CD13 were effectively enriched to approach saturation after 4 rounds of panning. The recovery rate in the fourth round was 30 times that in the first round. Twenty selected phages were verified by ELISA, and the signals of 10 phages were higher than the control. The sequences of the peptides P9 and P7 showed 83% and 100% identity with those of human cytomegalovirus (HCMV) UL38 and UL105, respectively. The peptides bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptides P9 and P7 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels. CONCLUSION: Two peptides (P7 and P9) that can specifically bind to CD13 have been screened successfully, and these two peptides show specific binding to CD13 on the membrane of THP-1 cells.

[Construction of recombinant plasmid pIRES2-EGFP/CCK and its expression in vivo and in vitro].

OBJECTIVE: To construct eukaryotic expression plasmid of porcine CCK gene pIRES2-EGFP/CCK and express it in COS-7 cells and hamsters. Methods The aimed segments were obtained from intermediate vector pMD18-T/CCK and were inserted into an eukaryotic expression plasmid pIRES2-EGFP to construct a recombinant expression plasmid pIRES2-EGFP/CCK. The recombinant expression plasmid was transfected into COS-7 cells by liposome-mediated gene transfer method and was observed through fluorescence microscope. The plasmid was injected into the skeletal muscle of hamsters directly to detect the expression of the recombinant plasmid in vivo. RESULTS: A recombinant eukaryotic expression plasmid pIRES2-EGFP/CCK was successfully constructed. Green fluorescent protein could be detected in the transfected COS-7 cells 24, 48, and 72 hours after the transfection. On the 4th day postinjection into the skeletal muscle of hamsters, the protein could be detected at the injection site and the fluorescence intensity became much stronger on the 14th day than that on the 4th day. On the 42nd day the protein level increased. The green fluorescence protein was never expressed in the untransfected cells. CONCLUSION: The porcine CCK gene eukaryotic expression plasmid pIRES2-EGFP/CCK is constructed successfully, and is expressed in mammal COS-7 cells and hamsters in vivo. The research paves the way for the cross immunity therapy of hamster pancreatic carcinoma.

[Construction of recombinant plasmid pIRES2-EGFP/CCK and its expression in vivo and in vitro].

OBJECTIVE: To construct eukaryotic expression plasmid of porcine CCK gene pIRES2-EGFP/ CCK and express it in COS-7 cells and hamsters. METHODS: The aimed segments were obtained from intermediate vector pMD18-T/CCK by the method of restricted enzymatic resection and were inserted into a eukaryotic expression plasmid pIRES2-EGFP to construct a recombinant expression plasmid pIRES2-EGFP/CCK. The recombinant expression plasmid was transfected into COS-7 cells by liposome-mediated gene transfer method and observed through Fluorescence microscopy. The plasmid was injected into the skeletal muscle of hamsters directly to detect the expression of the recombinant plasmid in vivo. RESULTS: A recombinant eukaryotic expression plasmid pIRES2-EGFP/CCK was successfully constructed. Green fluorescent protein could be detected in the transfected COS-7 cells 24, 48, and 72 hours post transfection and the expression of green fluorescent protein reached its peak 72 h post transfection. The green fluorescent protein could be detected at the injection site on the 4th day post injection and the fluorescence intensity became stronger on the 14th day. The level of fluorescence became ever stronger on the 42nd day. No expression of green fluorescence was detected in the control group. CONCLUSION: Porcine CCK cDNA eukaryotic expression plasmid pIRES2-EGFP/CCK has been successfully constructed and expressed in mammal cells COS-7 and hamster in vivo. The research paved the way for cross immunity therapy of hamster pancreatic carcinoma.

Hepatic volumetry with PhotoShop in personal computer.

BACKGROUND: Convenient way to clarify liver volume or tumor volume in the liver is eagerly demanded by hepatobiliary surgeons, for so many aspects of clinical work need to know the liver volumetry. At present, some methods have been used to measure the liver volumetry, such as computed tomography (CT) scans, three-dimensional ultrasound volumetric system([1]) and 3-dimensional sonography([2,3]) et al. But enough volumetric information was failed to obtain by surgeons and a new way of measuring the liver volumetry that can be operated by themselves is exigent. Whereas we devise a new method of using PhotoShop in personal computer to measure the liver volumetry. METHODS: A piece of whole CT film was transformed to a high quality digitized image by digital camera or scanner and then the digitized image was conducted as JPEG file into personal computer. The JPEG image file of CT film was opened by PhotoShop. Determining the edge of interested areas, and the data of pixel values of the interested areas divided by 1 cm2 pixel value will produce the actual area with the unit of square centimeter. If section thickness of CT scan is 1 cm, the sum of the areas of the liver or tumor in all sections naturally is the volume of the liver or tumor. RESULTS: Comparison of 10 hepatic volumes gained by this method and those gained by the GE Prospeed CT set showed a good relativity between the two groups. The volumes of three right lobes were calculated by this method before lobectomy and their real volumes were obtained postoperatively by a volumenometer. Their variation was limited to 5%. CONCLUSIONS: Hepatic volume obtained by PhotoShop is reliable. This method can be used to measure hepatic volume perfectly to meet clinical demand, and many parameters such as liver resection rate, graft volume can be achieved. The disadvantage of this method is the step of copying the pixel value from PhotoShop to Microsoft Excel.