Purpose To investigate the mutation status of KRAS and PIK3CA gene in colorectal cancer (CRC) primary lesions and corresponding liver metastasis and its clinical significance.Methods The gene mutations of KRAS and PIK3CA were detected in 58 cases of primary lesions of CRC and corresponding liver metastasis tissue by real-time PCR.Results The mutation rates of KRAS were 31.03％ (18/58) and 25.86％ (15/58) in primary lesions of CRC and corresponding liver metastasis tissue,respectively,in which G12D was most commonly detected.The mutation rates of PIK3CA were 8.62％ (5/58) and 10.34％ (6/58) respectively,in which the most common mutation site was E545K.Only one case carried simultaneously both mutations of KRAS (G12D) and PIK3CA (E545K).The mutation of KRAS and PIK3CA had a good consistency between primary lesions and liver metastasis.Univariate analysis showed that the mutation of KRAS was related to the primary lesion of tumor location,the quantity of metastasis and the types of tumor (P ＜ O.05),PIK3 CA mutation was associated with the synchronous/metachronous liver metastasis and the quantity of metastasis (P ＜ 0.05).Multivariate Cox regression analysis showed that synchronous/metachronous liver metastasis and the mutation of KRAS were influencing factors for prognosis of CRC.The overall survival of patients with CRC who had simultaneous liver metastases was longer than those with heterotopic liver metastases;the overall survival of KRAS wild-type mutant patients was longer than those of mutant patients (P ＜ 0.05).Conclusion The G12D site of KRAS gene has the highest mutation frequency in CRC,KRAS/PIK3CA mutation has a good consistency of the primary lesions of CRC and corresponding liver metastasis.Primary lesions can be as the source of molecular detection.To achieve individualized treatment,we need to reassess the genetic status of metastasis based on the choice of targeted therapy for precision medicine.

Objective To explore the methods of Y-V vascular pedicle lengthening technique for cover-age of the defect of great toe after wrap-around flap transfer, and decrease the morbidity of donor site in great toes. Methods Fifteen patients received three kinds of flap for immediate resurfacing of donor defect of the great toes during wrap-around flap transfer, all flaps were harvested with Y-V vascular pedicle lengthening technique. Among them, 9 cases received the free lateral tarsal flaps transplantation for coverage of defect in donor great toes, 5 cases was repaired by medial tarsal flaps, and only one was treated by the medial plantar flap. Results All the flaps survived postoperatively. The medial plantar flap encountered the venous crisis in postoperative 3 days, and regained the normal blood supply after continuous bleeding for 2 days. All patients were satisfied with cosmetic and functional outcome in average 10- month follow-up. The appearance and sensory function of donor toe repaired by medial tarsal flaps was best among three ones. Conclusion Depending on the Y-V vascular pedicle lengthening technique, the local pedical flaps of foots are enlarged scope for coverage of great toe after wrap-around flap harvest, which can decrease the complication of donor site at the most.

Objective To explore the experience in treatment of primary retroperitoneal tumors (PRPT).Methods The clinical data of 62 cases with primary relroperitoneal tumors from 2001 to 2007were studied retrospectively.Results The abdominal mass was the main symptom in PRPT.There were 29 cases of benign tumors and 33 cases of malignanat tunlors respectively.Complete resections were performed in 47 patients(76%),partial resection was conducted for 10 patients.Resection could not be performed in 5 patients so only biopsy was conducted,12 patients accepted combined organ resections,2 patients died.Conclusion Adequate preoperative preparation and excellent surgical technique are the keys to compele tumour resection.

Objective To reveal the role of serum ACE2/Ang (1-7)in the occurrence of atrial fibrillation (AF)and find new targets for the prevention and treatment of AF by analyzing the correlation between the serum concentration of ACE2/Ang (1-7 )in patients with rheumatic valvular heart disease and the occurrence of AF. Methods We collected the basic clinical information and peripheral venous blood of patients with rheumatic heart valve disease (totally 46 patients,including 24 with AF and 22 with SR).ELISA method was used to detect the serum concentration of ACE2,Ang (1-7)and AngⅡ in the serum samples.Then the differences and correlation between the two groups were analyzed.Results In the AF group ① the diameter of the left atrium was significantly greater than that in the SR group [(60.70±3.08 vs.48.15±2.16)mm,P<0.05];② the serum concentration of AngⅡ was significantly higher than that in the SR group [(45.88±2.87 vs.35.78±1.08)pg/mL, P<0.05],AngⅡ and left atrium diameter were positively correlated (Pearson test,P<0.05);③ the serum concentrations of ACE2 [(7.87±0.74 vs.11.65±0.57)U/L,P<0.05]and Ang (1-7)[(146.05±17.61 vs. 321.71±36.50)pg/mL,P<0.05]were significantly lower than those in the SR group,and negatively correlated with left atrium diameter (Pearson test,P<0.05);④ the serum concentration of Ang (1-7)was negatively correlated with AngⅡ concentration (Pearson test,P<0.05).Conclusion For patients with rheumatic valvular heart disease,ACE2/Ang (1-7 )may play a protective role in the occurrence of AF via antagonizing AngⅡ and inhibiting atrial remodeling.

AIM:To study the effects of extracellular potassium on the protein expression of wild-type HERG and its mutant L539fs/47.METHODS:Wild-type HERG (WT) or its mutant HERG-L539fs/47 (MT) were transfected into HEK293 cells for 36 h.The cells were incubated in different media containing 0.8, 4.3 or 10 mmol/L potassium.Af-ter 6 h of incubation, the protein expression of HERG was detected by flow cytometry.After 12 h of incubation, the locali-zation and quantity of the proteins were detected by laser confocal imaging and Western blot.RESULTS: Different from the retention of mutant protein in cytoplasm, wild-type HERG protein was mainly distributed in the cell membrane.The 2 proteins both increased with the changes of extracellular potassium.Flow cytometry showed that the fluorescence in the 2 groups both increased with the changes of extracellular potassium ( P<0.01 ) .The fluorescence in WT group was signifi-cantly higher than that in MT group (P<0.01).Western blot showed that mutant HERG protein included only one 60 kD band, different from the 135 kD and 155 kD bands in wild-type HERG, which were affected by the changes of extracellular potassium (P<0.05).CONCLUSION:The retention of HERG mutant L539fs/47 protein in the cytoplasm is more than wild-type HERG.Chronic high extracellular potassium keeps the stability of wild-type and mutant HERG proteins on the cell membrane.Chronic low potassium reduces the expression of HERG channel proteins in a time-dependent manner.

AIM: To explore the influence of Huqi San on the Hedgehog signaling pathway in rats with prehe-patocarcinoma.METHODS: The model of prehepatocarcinoma in the rats was established by a modified solt-farber method.The rats were intragastric administrated with Huqi San solution for 3 d after subtotal hepatectomy.Four weeks after administration of the Huqi San solution, the hepatic damage was observed by histopathological analysis.The protein expression of glutathione S-transferase-π (GST-π), alpha-fetoprotein (AFP), OV6, albumin (ALB) and glioma-associated oncogene homolog 2 (Gli2) was detected by immunohistochemistry and immunofluorescence staining.The expression of Sonic hedgehog (Shh), Smoothened (Smo), Gli2, cyclin D and cyclin E at mRNA and protein levels in the rats was determined by RT-qPCR and Western blot, respectively.The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyltransferase (GGT) were assayed using diagnostic kits.RESULTS: Compared with model group, Huqi San decreased the serum levels of ALT, AST and GGT, and alleviated the pathological changes in prehepatocarcinoma rats.Huqi San inhibited the protein expression of GST-π and AFP (P<0.05) in the prehepatocarcinoma rats.Huqi San also promoted the protein expression of OV6 and ALB (P<0.05).Furthermore, Huqi San activated Hedgehog signaling pathway and its downstream targeting molecules such as Shh, Smo, Gli2, cyclin D and cyclin E.In addition, the results in vitro showed that Huqi San may activate Hedgehog signaling pathway and promoted oval cell proliferation.CONCLUSION: Huqi San not only promotes hepatic progenitor cell proliferation, but also induces hepatic progenitor cell differentiation and inhibits prehepatocarcinoma in the rats probably via activating Hedgehog signaling pathway.

Objective To evaluate the diagnosis and surgical management for pancreatic duct stones (PDS). Methods The clinical data of 19 PDS patients admitted to our hospital from Jan. 1985 to Sep. 2003 were analyzed retrospectively. Results Epigastric pain was the first symptom in all patients,and chronic pancreatitis was associated, pancreatic carcinoma coexisted in 6 cases. Fifteen cases were found to have PDS or pancreatic duct dilation by B-type ultrasonography. Twelve patients underwent pancreatolithotomy plus side to side Roux-en-Y pancreaticojejunostomy. Pancreatoduodenectomy was performed on four patients ,caudal pancreatectomy and pancreatolithotomy on three patients. Six pancreatic carcinoma patients died during follow-up of 9～26 months. Abdominal pain was relieved completely in 11 cases and partially in 1,stones and pain recurred in one patient. Conclusions B-type ultrasonography、CT and endoscopic retrograde cholangiopancreatography(ERCP) are most useful diagnostic methods;Surgical procedures should be selected according to the following factors: (1) location of the pancreatic duct stones. (2) stenosis of the main pancreatic duct.(3) pancreatic duct stones associated with or without pancreatic carcinoma.

To investigate the cytotoxicity of the homemade peptide cationic liposome CDO14 and its efficacy of RNA interference (RNAi). MTT method was used to determine the cytotoxicity of the liposome to a human lung cancer cell line Luc-A549 that can express luciferase stably. Luciferase siRNA (Luc-siRNA) was transfected into Luc-A549 cells by CDO14. Contents of luciferase in the transfected cells were detected by luminous instrument and contents of total protein in these cells were detected by BCA method. Nude mice were inoculated with Luc-A549 cells in axilla to establish xenograft tumor model. Complexes of Luc-siRNA and the cationic liposomes were injected into the modeling mice via tail vein. Contents of luciferase in the transfected mice were detected by the whole body imaging system. The cytotoxicity of the homemade cationic liposome was similar to that of commercial liposome DOTAP, and lower than that of Lipo2000. The siRNA transfection efficacy mediated by CDO14 was higher than that mediated by DOTAP. The homemade peptide cationic liposome CDO14 is expected to serve as delivery vector in gene therapy because of its low cytotoxicity and high transfection efficiency.
Animals ; Cations ; Cell Line, Tumor ; Fatty Acids, Monounsaturated ; Genetic Therapy ; Genetic Vectors ; Humans ; Liposomes ; Luciferases ; Lung Neoplasms ; Mice ; Mice, Nude ; Peptides ; Quaternary Ammonium Compounds ; RNA Interference ; RNA, Small Interfering ; Transfection

Objective To explore the clinical value of serum procalcitonin (PCT)-based antibiotic therapy in the second-classedexacerbations of chronic obstructive pulmonary disease (AECOPD). Methods 240 patients diagnosised as AECOPD were randomized to the PCT group and the control group. Serum PCT levels of patients from the PCT group were measured 1 h after hospitalized and the third, fifth, eighth day respectively. When PCT < 0.1 μg / L, patients will stop taking antibiotics and initiated while PCT≥0.1 μg / L. Antibiotic treatment in the control group was based on guidelines of COPD diagnosis and treatment. Results Duration of antibiotic therapy and hospitalization were respectively 5.6 ± 1.4 and 8.2 ± 1.1 days in the PCT group, 9.2 ± 2.2 and 11.4 ± 2.5 days in the control group (both P < 0.05). Mean costs of hospitalization expensesand antibiotic therapy were 5700 ± 201 and 1650 ± 189) yuan in the PCT group, 6210 ± 220 and 2350 ± 210 yuan in the control group (both P < 0.05). The clinical effective rate, times of exacerbation, one-year ΔFEV1, the 1-year hospitalization rate and time to next exacerbation all showed no significant differences between the two groups. Conclusion PCT-guided antibiotic treatment reduces antibiotic use inthe second-classed acute exacerbations patients.

OBJECTIVETo explore the effect of body mass index (BMI) on postoperative short-term prognosis and survival rate of gastric cancer patients.

METHODSClinical and follow-up date of 153 gastric cancer cases undergoing radical operation in our hospital from January to June 2010 were retrospectively analyzed. According to BMI, patients were divided into low group (BMI<18.5, 23 cases), normal group (18.5≤BMI<25.0, 95 cases) and high group (BMI≥25.0, 35 cases). Clinicopathological features and outcomes were compared the among three groups.

RESULTSAmong three groups, the differences in operation time, intraoperative blood loss, number of lymph node retrieved, postoperative hospital stay, lymph node metastasis rate, tumor staging and postoperative complication morbidity were not statistically significant (all P>0.05). Preoperative hemoglobin in the low group was significantly lower as compared to normal and high groups [(106.1±13.8) g/L vs. 113.5±5.2) g/L and (123.5±8.7) g/L, F=3.265, P=0.041], and so was the preoperative albumin [(38.7±2.5) g/L vs. (41.3±0.8) g/L and (43.5±1.4) g/L, F=8.516, P=0.000]. The ratio of gastric cardiac cancer in the low group was significantly lower as compared to the normal and high groups[34.8%(8/23) vs. 68.4%(65/95) and 62.9%(22/35), χ(2)=8.913, P=0.012]. Five-year survival rate of the low, normal and high groups were 43.5%, 50.5% and 65.7% respectively(P=0.189). Subgroup analysis showed that the 5-year survival rate of patients with gastric cardiac cancer in the low group was significantly lower as compared to those in the high group (25.0% vs. 84.6%, P=0.004).

CONCLUSIONSBMI dose not generally play a role in short-term outcomes and long-term survival of the gastric cancer patients. Nutritional improvement and body weight maintenance may be beneficial to low BMI patients, especially those with gastric cardiac cancer.

Blood Loss, Surgical ; Body Mass Index ; Gastrectomy ; Humans ; Length of Stay ; Lymph Nodes ; Lymphatic Metastasis ; Neoplasm Staging ; Operative Time ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Survival Rate ; Treatment Outcome