Objective To discuss the advance and experience of postoperative management in multivisceral transplantation(MVT).Methods Five patients with abdominal multiple organs malig- nances were subjected to multivisceral transplantation.The excised specimens included the liver,pan- creas,duodenum,stomach,spleen,mesentery and all of the involved lymph nodes.The transplanted organs were liver,pancreas,duodenum and parts of intestine.Patients were monitored and sustained stably in breath,hemodynamics and homoeostasis.The coagulation was also monitored and kept in balance.The intensive immunosuppressive protocol,including steroid,tacrolimus,mycophenolate mofetil and Zenapax,was used to prevent the acute rejection.The endocrine and exocrine of pancreas were tested,color Doppler examination was done every day,and Somatostatin was used to prevent pancreatitis.Drugs to bacteria,fungus and virus infection were prophylactically used.Results All patients waked up with 4-6 h after operation and began recovering with 3-7 days.Dynamic examina- tions revealed that the functions of grafted liver,pancreas and duodenum were satisfactory.No com- plications of acute rejection,pancreatitis,bleeding and thrombosis occurred.Conclusion Many ques- tions in postoperative management related to multivisceral transplantation are devoted to study.Nice experience of management to organ transplantation,detailed monitoring and prophylactically medica- tions are safeguards to steady recovering and further prognosis.

Objective To explore the feasibility of anti-85B and ESAT-6 monoclonal antibodies targeted contrast agent of CT by the murine acute tuberculosis animal model.Methods Preparation the targeted contrast agent of computed tomography by iodine atoms coupled with anti-85B and ESAT-6 murine monoclonal antibodies after purified.Calculate the label rate and the quality of 127Ⅰ of the targeted contrast agent solution,and dilute the contrast agent solution to the required concentration (5μg I/ml) to spare.There were twenty mice of acute tuberculosis animal model,which were divided into four groups by completely randomized digital table and each group was five animals.According to the different antibody named as 85B group and ESAT-6 group of targeted contrast agent,common contrast agent and blank control separately.The common contrast agent group was injected with diluents of iohexol,which was diluted into the same concentration with the targeted contrast agent.The control group was injected with antibody diluents pH 7.4 Phosphate Buffered Saline (PBS).All the animals were scanned before and after injection the contrast agent in different time,such as immediate,6 hours,12 hours and 24 hours.Observe the display and changes of the murine tuberculosis lesions,and measurement the CT value,which was regarded as evaluating mark.Enhancement ratio was also calculated.Two sample mean differences with t test and the multiple sample mean differences with ANOVA.Results The volume of anti-85B contrast agent solution was 2.52 ml,and the quality of antibody and 127Ⅰ were range from 210 to 255 μg and 10.5 to 16.6 μg respectively.The volume of anti-ESAT-6 contrast agent solution was 2.93 ml,and the quality of antibody and 127Ⅰ were 147 μg and 20.58 μg respectively.The lesions of the control group showed no visible density changes before and after injection of PBS.The CT value of the lung lesions in the targeted contrast agent group were gradually increased with time,and the lesion showed visible enhancement after the contrast injection twelve hours(t12),and also remained visible enhancement after injection twenty-four hours(t24)[85B group t12 =(-125.04 ± 13.58) HU,t24 =(-117.37 ± 12.28) HU and ESAT-6 group t12 =(-122.14 ± 19.01) HU,t24 =(-114.23 ± 17.08) HU],which is significant difference compared to the common contrast agent [t (85B-24 h) =4.05,t (ESAT-6-24 h) =6.39,P ＜ 0.05].Conclusions The targeted property of anti-85B and ESAT-6 murine monoclonal antibody contrast agent of CT had been partly proved by the acute tuberculosis animal model.Also provided an experimental basis for further study of tuberculosis targeted contrast agent.

Objective To investigate the effects of Krüppel like factor 4 (KLF4)on matrix metalloproteinase 9 (MMP9)ex-pression in hepatocellular carcinoma(HCC).Methods A total of 50 primary hepatocellular carcinoma samples and their correspond-ing adjacent tissues specimens were collected.The expression of KLF4 and MMP9 were detected by IHC,Western blot and qRT-PCR.After KLF4 gene was transfected into hepatocellular carcinoma cell line (HepG2 cell line),the expressions of KLF4 and MMP9 were conformed by qRT-PCR and Western blot.Migration and invasion of HepG2 cell line transfected by KLF4 were detec-ted by wound-healing assay and invasion assay.Results Compared to corresponding adjacent tissues,The expression of KLF4 was significantly lower in HCCs(P <0.05),and MMP9 expression was remarkably higher in HCCs(P <0.05).KLF4 over-expression inhibited the expression of MMP9 on the protein and mRNA levels.Wound-healing assay and invasion assay confirmed that KLF4 regulated cell invasion and migration through regulating MMP9 expression.Conclusion KLF4 showed low expression in HCCs,and MMP9 was overexpressed.Up-regulation of KLF4 could decrease the expression of MMP9 in HepG2 cell line,which inhibited inva-sion and migration.

This study aimed at investigating the effects of the extract of Chinese herb,Nansheteng (C.orbiculatus Thunb.),on human HepG2 cells through the overexpression of mTOR.The GV238-mTOR recombinant plasmids were transfected into HepG2 cells using molecular biological technology.The expression level of mTOR was evaluated by means of relative activity of luciferase and western blot.Human hepatic carcinoma HepG2/mTOR++ cells were treated with C.orbiculatus extract in different concentrations (20,40,80,160 and 320 tg·mL-1) for 24 h.The mTOR protein expression was detected by western blot.It was found that the protein expression of mTOR in transfected HepG2 cells was significantly enhanced.C.orbiculatus extract significantly inhibited the proliferation of HepG2/mTOR++ cells.Simultaneously,C.orbiculatus extract inhibited mTOR at its protein level in a dose-dependent manner.In conclusion,we successfully constructed recombinant mTOR cloning vectors,and established the stable HepG2 cell line with the overexpression of mTOR.Besides,C.orbiculatus extract significantly inhibited mTOR protein expression in human hepatic carcinoma HepG2 cells.

[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.

OBJECTIVE:To study the technical conditions and parameters of DiaionHP-20 macroporous adsorption resin for the purification of the anti-PMOP(primary Type I osteoporosis) active fraction from Camellia semiserrata seed.METHODS: With the elution ratios and purity of total flavonoids and polyphenol from Camellia semiserrata as indexes,the technical parameters of DiaionHP-20 macroporous adsorption resin including the adsorption quantity,the loading sample,the elution concentration and volume of alcohol were investigated.RESULTS: The adsorption quantity of DiaionHP-20 macroporous resin on total flavonoids and polyphenol were 59.5 mg?g-1 and 23.9 mg?g-1 respectively.The optimal technical conditions were as follows:the quantity of drug was 10.5 times that of resin;water-soluble foreign substance was washed off with distilled water weighing 3 times the weight of resin,and total flavonoids and polyphenol were eluted using 25% ethanol weighing 13.5 times the weight of resin.CONCLUSION:DiaionHP-20 macroporous resin had good purification effects on total flavonoids and polyphenol from Camellia semiserrata seeds.The process is simple,low cost,and suitable for lavge-scale production.

Objective To investigate the clinical value of a new anvil inserting method for esophagogastrostomy or esophagojejunostomy during laparoscopic radical proximal gastrectomy or radical total gastrectomy for gastric cancer.Methods The clinical data of 21 patients with gastric cancer who received laparoscopic radical proximal gastrectomy or radical total gastrectomy at the Southwest Hospital from March 2010 to February 2011 were retrospectively analyzed.Five trocars were inserted through the abdominal wall of the patients.After perigastric lymphadenectomy and mobilization of esophagus,an incision was made on the esophagus above the tumor,and then the anvil with drawn wire attached was inserted into the esophagus.An endo-cutter was applied to cut the esophagus adjacent to the incision left the drawn wire untouched,and then the stem of the anvil was pulled out by the drawn wire for laparoscopic anastomosis. Results The operations were successfully accomplished under the laparoscope with no conversion to open surgery.Fifteen patients received laparoscopic radical total gastrectomy and 6 received laparoscopic radical proximal gastrectomy. The mean operation time,volume of blood loss,time to off-bed activity,passage of flatus and postoperative duration of hospital stay were (257 ± 38) minutes,( 119 ± 32) ml,(2.5 ± 0.5 ) days,( 3.7 ± 0.8 ) days and (7.5 ± 2.6) days,respectively.No perioperative mortality,anastomotic bleeding or anastomotic fistula was detected.One patient was complicated with pulmonary infection + pleural effusion and was cured by conservative treatment; 1 was complicated with anastomotic stenosis which was alleviated by gastroscopic balloon dilation; 1 was complicated by incisional infection and was cured by medical treatment after drainage.No cancer cells were detected at the anastomotic ring or resection margin of the specimen.There were 4 patients with well-differentiated adenoma,8 with moderate-differentiated adenoma and 9 with poor-differentiated mucinous adenoma.There were 5 patients in stage Ⅰ,10 in stage Ⅱ and 6 in stage Ⅲ (UICC staging).Twenty-one patients were followed up for a mean period of (11 ±4) months (range,6-17 months ),no tumor recurrence or metastasis was detected. Conclusions The new technique for anvil insertion is safe,effective and easy for manipulation and learn.It offers a new approach for laparoscopic digestive tract reconstruction.

Advanced gastric cancer is usually dealt with D2 radical dissection. There are different opinions as to whether it is necessary to perform D3 radical lymphadenectomy.Some scholars thought that properly enlarged radical dissection can improve long-term outcomes for the treatment of advanced gastric cancer.In recent years,laparoscopic D1 and D2 radical dissection of gastric cancer could be carried out in many hospitals.However,the technique and related skills for performing D3 radical lymphadeneetomy through laparoscope remains to be explored.Based on our previous experiences,D3 radical lymphadeneetomy using artery suspension method and medial-to-lateral approach for advanced gastric cancer is proved to be safe and feasihle.

Objective To compare the effects and significance of laparoscopic and open D2 gastrectomy on the expression of interleukin (IL)-6 and IL-10.Methods The clinical data of 146 patients with gastric cancer who were admitted to the Southwest Hospital from November 2010 to October 2011 were prospectively analyzed.All the patients were randomly divided into the laparoscopic group (75 patients) and open group (71 patients)according to the sealed envelope method.Laparoscopic or open D2 gastrectomy were performed according to the 14th edition of gastric cancer treatment guidelines of Japan Gastric Cancer Association.Peritoneal lavage fluid was collected at the beginning and the end of operation,and the concentrations of IL-6 and IL-10 in the peritoneal lavage fluid were detected by enzyme linked immunosorbent assay.The measurement data were analyzed using the t test,and the count data were analyzed using the chi-square test.Results The preoperative concentrations of IL-6 in the laparoscopic group and the open group were (34 ± 13)μg/L and (35 ± 12)μg/L,respectively,with no significant difference between the 2 groups (t =-5.110,P ＞ 0.05).The postoperative concentrations of IL-6 in the laparoscopic group and the open group were (4015 ± 1592)μg/L and (6724 ± 2112)μg/L,respectively.The postoperative concentration of IL-6 in the laparoscopic group was significantly lower than that of the open group (t =-8.367,P ＜ 0.05),and the postoperative concentrations of IL-6 were significantly higher than those before operation in the laparoscopic group and open group (t =-59.065,-87.123,P ＜0.05).The preoperative concentrations of IL-10 in the laparoscopic group and the open group were (43 ±9) μg/L and (42 ± 10) μL,respectively,with no significant difference between the 2 groups (t =1.190,P ＞0.05).The postoperative concentrations of IL-10 in the laparoscopic group and the open group were (92 ± 32)μg/L and (62 ± 23)μg/L,respectively.The postoperative concentration of IL-10 was significantly higher than that of the open group (t =6.408,P ＜ 0.05),and the postoperative concentrations of IL-10 were significantly higher than those before operation in the laparoscopic group and the open group (t =-12.680,-6.802,P ＜ 0.05).Conclusion Peritoneal inflammatory reaction is relatively lighter after laparoscopic D2 gastrectomy when compared with open D2 gastrectomy,which might prevent the peritoneal metastasis of gastric cancer mediated by IL-6.

Although the surgical procedure and approach of da Vinci robotic surgical system-assisted radical resection of gastric cancer are gradually mature,it is rarely used for the resection of gastric stump cancer because of the complexity and low resection rate.In November of 2012,resection of gastric stump cancer using da Vinci robotic surgical system was performed in the Southwest Hospital.The short-term efficacy was satisfactory after the follow-up for 12 months.Da vinci robotic surgical system has the advantages of clear vision,easy manipulation of abdominal adhesion detaching,flexible operation and stable traction during resection of gastric stump cancer.