Cytotoxic activity of gypenosides and gynogenin against non-small cell lung carcinoma A549 cells.

The activity of gypenosides and gynogenin of Gynostemma pentaphyllum against non-small cell lung carcinoma (NSCLC) A549 cells was investigated to identify the structural characteristics of gypenosides and gynogenin to have anti-NSCLC activity. Of the tested dammarane-type compounds, 20S-dammar-24-en-2α,3ß,12ß,20-tetrol showed the strongest activity against A549 cells. Based on the structure and cytotoxic activity relationships of gypenosides and gynogenin, the OH group in C-2, the connected sugar number and the configuration in C-20 were important for cytotoxic activity against A549 cells.

Novel dammarane saponins from Gynostemma pentaphyllum and their cytotoxic activities against HepG2 cells.

Two new dammarane saponins, 2α,3ß,12ß-trihydroxydammar-20(22),24-diene-3-O-[ß-D-glucopyranoxyl(1→2)-ß-D-6″-O-acetylglucopyranoside (1, namely damulin C) and 2α,3ß,12ß-trihydroxydammar-20(21),24-diene-3-O-[ß-D-glucopyranoxyl(1→2)-ß-D-6″-O-acetylglucopyranoside (2, namely damulin D), were isolated from the ethanol extract of Gynostemma pentaphyllum, which had been heat processed by steaming at 125 °C. The NMR spectroscopic data of the novel saponins were completely assigned by using a combination of 2D NMR experiments including (1)H-(1)H COSY, HSQC, and HMBC. Their cytotoxic activities of human liver adenocarcinoma HepG2 cells were evaluated in vitro. They showed cytotoxicities against HepG2 cell line with IC50 of 40±0.7 and 38±0.5 µg/ml, respectively.

Clinicopathological characteristics of intrahepatic cholangiocarcinoma in patients with cirrhosis.

BACKGROUND/AIMS: The purpose of this study was to explore the clinicopathological characteristics of intrahepatic cholangiocarcinoma (ICC) in patients with cirrhosis. METHODOLOGY: A total of 155 patients with ICC were divided into those with cirrhosis (n=52) and those without cirrhosis (n=103). We compared the clinicopathological features of patients in both groups. RESULTS: The prevalence of HBsAg seropositivity and hepatolithiasis in ICC patients with cirrhosis was higher than that in patients without cirrhosis. Compared with noncirrhotic patients, cirrhotic patients had a higher incidence of reduced albumin (46.1% vs. 25.2%, p<0.008) and elevated total bilirubin (TBIL) levels (44.2% vs. 24.3%, p=0.011). The resectability rate in cirrhotic patients was lower than that in noncirrhotic patients (63.7% vs. 80.6%, p=0.033). CONCLUSIONS: Among ICC patients, we found marked differences in clinicopathological characteristics and therapeutic approaches between cirrhotic and noncirrhotic patients. ICC patients with cirrhosis may have poorer prognosis than those without cirrhosis.

Vector-based RNA interference against vascular endothelial growth factor-C inhibits tumor lymphangiogenesis and growth of colorectal cancer in vivo in mice.

BACKGROUND: RNA interference (RNAi) technology is emerging as a very potent tool to generate a cellular knockdown of a desired gene. The aim of this study was to explore whether RNAi targeting vascular endothelial growth factor-C (VEGF-C) could inhibit colorectal tumor lymphangiogenesis and tumor growth. METHODS: We used vector-based RNAi to inhibit VEGF-C expression in colon cancer in vitro and in vivo. VEGF-C expression was quantified by real-time polymerase chain reaction and Westen blotting. To establish LoVo cell tumor xenografts in mice, we subcutaneously inoculated 1.0 x 10(6) LoVo cells in nude mice; after injection, tumors were allowed to grow for 4 weeks until the volume reached (75.8+/-55.8) mm(3). The mice were then randomly divided into two groups: (1) the VEGF-C-siRNA group (n=10) received direct injection of "therapeutic" plasmid 50 microg of LoVo-VEGF-C-siRNA into the tumor mass; (2) the control group (n=10) were injected with LoVo-control in 20 microl of sterile 0.9% NaCl solution into the tumor mass. Tumor growth, microlymphatics and microvessels were compared for mice administered either systemic VEGF-C-siRNA or control over 4 weeks. RESULTS: The mRNA and protein expression of VEGF-C in the colon tumor cell line, LoVo, stably transfected with a VEGF-C-siRNA vector, were significantly downregulated 45.3% and 35.3% respectively. In vivo, four weeks after injection, the tumor volume were significantly smaller in VEGF-C-siRNA group than in LoVo-control group ((314.8+/-54.8) mm(3) vs (553.9+/-90.1) mm(3)); the incidences of lymph node metastasis (30%) in VEGF-C-siRNA were significantly inhibited compared with LoVo-control group (70%); in VEGF-C-siRNA group, the number of microlymphatics per microscopic field was (5.3+/-0.7) and the number of microvessels per microscopic field was (24.2+/-6.5) decreased compared with LoVo-control group (12.5+/-6.9) and (42.1+/-7.4) (all P<0.001). CONCLUSION: Inhibition of VEGF-C expression using siRNA-mediated gene silencing vectors reduced lymphangiogenesis and lymph node metastasis and enhanced survival.

[Breast intraductal lesion resection under breast fiberoptic ductoscopy].

OBJECTIVE: To explore the feasibility and to sum up the experience of breast intraductal neoplasm resection under breast fiberoptic ductoscopy (FDS). METHODS: FDS was performed on 548 patients with nipple discharge from Sep.2004 to Nov.2006. The clinical data of breast intraductal neoplasm found by FDS in patients who underwent tumor resection were analyzed, and the breast intraductal neoplasm image characteristics, diagnosis, operative type and postoperative pathological results were analyzed. RESULTS: Of the 548 patients with nipple discharge, intraductal neoplasm was found in 187 cases (34.1%), intraductal papilloma in 159 cases (29.0%), intraductal papillomatosis in 12 cases (2.2%), and breast carcinoma in 16 cases (2.9%). One hundred thirty-five patients were operated on in our hospital, of whom 91 were performed tumor resection or segmentectomy under the localization by FDS, and the other 44 were performed segmentectomy after breast duct infusion of methylene blue. The diagnostic rate under FDS in the FDS group (97.8%) was higher than that in the breast duct infusion methylene group (86.4%) (chi2=6.96, P=0.008). CONCLUSION: FDS is not only an accurate diagnosis for breast intraductal lesion, but also an assistance to localize the breast intraductal neoplasm and to remove them in the operation.

Metabolite profiling of gypenoside LVI in rat after oral and intravenous administration.

Gypenoside LVI, one of the major bioactive triterpene saponins in Gynostemma pentaphyllum, has been regarded as a potential and promising lead drug for anti-tumor strategy. To better understand the pharmacological activities of the component, an investigation of its in vivo metabolism is important and necessary. In the present study, a liquid chromatography-ion trap time of flight tandem mass spectrometry has been utilized to discover and identify the metabolites of gypenoside LVI in rat urine after oral and intravenous administration. Negative electrospray ionisation mass spectrometry was used to discern gypenoside LVI and its possible metabolites in urine samples. As a result, after oral and intravenous administration, eight and six metabolites together with gypenoside LVI were detected and identified in rat urine, respectively. As metabolites of gypenoside LVI, they have never been reported before. Deglycosylation and dehydration were found to be the major metabolic processes of gypenoside LVI in rat.

Role of CXC chemokine receptor type 7 in carcinogenesis and lymph node metastasis of colon cancer.

Upregulated expression of the CXC chemokine receptor type 7 (CXCR7) promotes breast, lung and prostate cancer progression and metastasis. However, the role of CXCR7 in colon cancer has not been determined. We hypothesized that increased CXCR7 expression may contribute to human colon cancer occurrence and progression. Reverse transcription quantitative polymerase chain reaction and western blot analysis were performed on 34 malignant and 18 normal colon tissue specimens. The specimens were obtained from 19 male and 15 female patients, with a mean age of 52 years (range, 34-79 years). Of the 34 patients, 20 had lymph node metastases. None of the patients had received adjuvant radiotherapy or chemotherapy prior to surgery. This study demonstrated that CXCR7 levels were significantly higher in colon tumors compared with those in normal colon tissue (P﹤0.01). In addition, lymph node metastatic colon tumors exhibited significantly higher CXCR7 expression compared with non-metastatic tumors (P﹤0.01); however, there were no differences in CXCR7 expression among distinct histopathological types (well-differentiated vs. moderately-to-poorly differentiated adenocarcinoma, P﹥0.01). Therefore, the evidence obtained from the present study supports involvement of the upregulated CXCR7 expression in colon tumorigenesis and lymph node metastasis.

[Effect of Chinese traditional medicine mixture on inflammatory response in rats with severe burn].

OBJECTIVE: To investigate the regulatory effect of Chinese traditional medicine mixture (CTMM) on inflammatory response in rats with severe burn. METHODS: One hundred and ten rats were randomly divided into 3 groups:scalded rats inflicted by 30% III degree scald were treated with CTMM and SD-Ag (CTMM group), scalded rats inflicted by 30% III degree scald were treated with SD-Ag alone (scalded group), and healthy rats were treated with SD-Ag (normal group). The serum contents of TNF-alpha, IL-beta, IL-6, IL-8, IL-4, and IL-10 in rats in the 3 groups were dynamically monitored. RESULTS: The serum contents of TNF-alpha, IL-1beta, IL-6, IL-8, IL4, and IL-10 evidently increased in both the CTMM and scalded groups. But the contents of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6, and IL-8) in the CTMM group were much lower than those in the scalded group. However, the contents of anti-inflammatory cytokines (IL-4 and IL-10) in the CTMM group were much higher than those in the scalded group. CONCLUSION: CTMM has double-way regulatory effect on the inflammatory response in rats with severe burn.

Major hepatectomy is a safe modality for the treatment of intrahepatic cholangiocarcinoma in selected patients complicated with cirrhosis.

OBJECTIVE: The objective of this paper is to evaluate the perioperative outcomes of major hepatectomy for intrahepatic cholangiocarcinoma (ICC) in patients with cirrhosis. METHODS: We retrospectively evaluated the preoperative, intraoperative, and postoperative findings in 42 consecutive patients with cirrhosis and in 102 patients with normal livers who underwent major hepatectomy for ICC. RESULTS: Preoperative liver function was worse in patients with cirrhosis compared to patients without cirrhosis. Cirrhotic patients had significantly higher intraoperative blood loss, longer operation time, and longer hospital stay than non-cirrhotic patients. However, the two groups had similar overall morbidity and hospital mortality rates and similar rates of liver failure or other complications. Their R0 resection rates, resection margin widths and disease-free survival rates were also similar. CONCLUSIONS: Major hepatectomy for ICC can be performed in selected cirrhotic patients with acceptable morbidity and mortality rates, as compared to patients without cirrhosis.

Metabolic profiling of Gynostemma pentaphyllum extract in rat serum, urine and faeces after oral administration.

Folk drug Gynostemma pentaphyllum (Thunb.) Makino contains many biologically active phytochemicals which have been demonstrated to be effective against chronic diseases. As in vivo anti-tumor experiments of G. pentaphyllum extract (GP) show much stronger antitumor activities than in vitro, it is important and necessary to understand the metabolic study of GP. A sensitive and specific U-HPLC-MS method was utilized for the first time to rapidly identify gypenosides and its possible metabolites in rat serum, urine, and faeces after oral administration. Solid phase extraction was utilized in the sample preparation. Negative Electrospray ionisation (ESI) mass spectrometry was used to discern gypenosides and its possible metabolites in rat samples. As a result, after oral administration, a total of seven metabolites of G. pentaphyllum extract were assigned, two from the rat serum and seven both from the rat urine and faeces. As metabolites of G. pentaphyllum extract, all of them have never been reported before.

Dammarane-type saponins from heat-processed Gynostemma pentaphyllum show fortified activity against A549 cells.

An ethanol extract from heat-processed Gynostemma pentaphyllum showed more potent cytotoxic activity against human lung adenocarcinoma A549 cells than that of raw G. pentaphyllum. Four constituents were isolated from heat-processed G. pentaphyllum using resin HP-20, silica gel and reversed ODS column chromatography. They were identified by mass and NMR spectra as damulin A and damulin B, gypenoside L and gypenoside LI, respectively. To evaluate the efficacy of these four constituents, the MTT cytotoxicity assay was performed using A549 cells. Based on the structure of these four constituents, the results indicate that the hydroxyl group in C-2 and double bond in C20(21) and C20(22) positions are of importance in inhibition of A549 cell proliferation.

Effect of CYP3A4*18B polymorphisms and interactions with OPRM1 A118G on postoperative fentanyl requirements in patients undergoing radical gastrectomy.

The present study aimed to investigate the effect of cytochrome P450 3A4 (CYP3A4)*18B polymorphisms and the interaction of the µ opioid receptor gene (OPRM1) A118G and CYP3A4*18B polymorphisms on postoperative fentanyl analgesia in Chinese Han patients undergoing radical gastrectomy. In total, 97 patients scheduled to undergo radical gastrectomy under general anesthesia were enrolled in this study. Post­operative patient­controlled intravenous analgesia of fentanyl was administered as analgesia up to 48 h following surgery. Venous blood (2 ml) was obtained from each patient to measure the OPRM1 A118G and CYP3A4*18B genotypes. The differences in fentanyl consumption and adverse effects were compared among the genotypes at 24 and 48 h following surgery. In the first 48 h following surgery, patients in the CYP3A4*18B/*18B group consumed significantly less fentanyl compared with patients in the *1/*1 group (P=0.032). With regards to the joint genetic effect, during the 48­h period, patients with AA and *1*18B polymorphisms received fewer fentanyl doses compared with those with AG and *1*1 (P=0.049), while patients with AG and *1*18B polymorphisms received significantly fewer fentanyl doses compared with those with AG and *1*1 (P=0.010), and patients with *18B*18B polymorphisms received significantly fewer fentanyl doses compared with those with AA and *1*1 (P=0.024) or those with AG and *1*1 polymorphisms (P=0.006). No correlation between OPRM1 A118G and CYP3A4*18B and postoperative nausea, vomiting and dizziness was found. Results demonstrated that 48 h following surgery, patients with the CYP3A4*18B/*18B genotype required less fentanyl than patients with the CYP3A4*1/*1 genotype to control pain. Additionally, the combined genotype of CYP3A4*18B and OPRM1 A118G may affect fentanyl doses administered for pain control, but not postoperative nausea, vomiting and dizziness.

Association between polymorphisms of XRCC1 Arg399Gln and XPD Lys751Gln genes and prognosis of colorectal cancer in a Chinese population.

We conducted this study to detect associations between XRCC1 Arg399Gln and XPD Lys751Gln genotypes and survival of colorectal cancer patients treated with 5-FU/oxalipatin chemotherapy. We included 289 Chinese patients with advanced colorectal cancer, who had received 5-FU/oxalipatin chemotherapy as first-line treatment from January 2005 to January 2007. All patients were followed up till Nov. 2011. Genotyping for XRCC1 Arg399Gln and XPD Lys751Gln polymorphisms was based upon duplex polymerase-chain-reaction with the PCR-RFLP method. In our study, we found the XRCC1 399 Gln/Gln genotype to confer significantly higher rates of response to chemotherapy when compared to the Arg/Arg genotype [OR (95% CI)=2.56(1.57-2.55)]. patients with the XPD 751 Gln/Gln genotype had significantly higher rates of response to chemotherapy [OR (95% CI)=1.54(0.87-2.65)] and those with the XRCC1 399 Gln/Gln genotype had a longer average survival time and significantly lower risk of death than did those with the Arg/Arg genotype [HR (95% CI)=0.66(0.36-0.95)]. Similarly, those carrying the XPD 751Gln/Gln genotype had 0.51-fold the risk of death of those with XPD 751Lys/Lys [HR (95% CI)=0.51(0.33-0.94)]. In conclusion, it is suggested that the XRCC1 Arg399Gln and XPD Lys751Gln polymorphisms should be routinely assessed to determine colorectal patients who are more likely to benefit from 5-FU/oxalipatin chemotherapy.

Multivariate analysis of molecular indicators for postoperative liver metastasis in colorectal cancer cases.

AIMS: To explore the relationship between various molecular makers and liver metastasis of colorectal cancer (CRC). METHOD: Using immunohistochemistry, protein expression of CEA, nm23, c-met, MMP2, COX- 2, VEGF, EGFR, and CD44 was assessed in 80 CRC cases. The Chi-square test and logistic regression were performed to analyze the relationship between these indicators and CRC liver metastasis. RESULTS: There were significant differences in expression of CEA, MMP2, CD44, VEGF and EGFR between the liver metastasis and non metastasis groups (P < 0.05); no significant differences were noted for nm23, c-met, and COX-2 expression. Logistic regression analysis showed that only CEA, VEGF, and EGFR entered into the regression equation, and had significant correlations with CRC liver metastasis (α inclusion= 0.10, α elimination = 0.15, R2 = 0.718). CONCLUSIONS: Combination detection of CEA, VEGF, and EGFR may be an effective means to predict CRC liver metastasis. Nm23, c-met, MMP2, COX-2, and CD44, in contrast, are not suitable as prognostic markers.

Prognostic value of cirrhosis for intrahepatic cholangiocarcinoma after surgical treatment.

BACKGROUND: The surgical outcome and prognostic factors of intrahepatic cholangiocarcinoma are not fully understood. This study aimed to establish the clinical significance of cirrhosis for prognosis in patients with intrahepatic cholangiocarcinoma after surgery. METHODS: One hundred fifteen patients with intrahepatic cholangiocarcinoma who underwent surgical resection between December 2001 and January 2008 were retrospectively analyzed. The prognostic significance of clinicopathologic factors including cirrhosis was assessed by univariate and multivariate analyses. RESULTS: Thirty-two of the 115 patients (28%) had liver cirrhosis. Complete tumor removal (R0 resection) was performed in 42 patients (75%). Overall median survival time was 21 months, with 1-, 3-, and 5-year actuarial survival rates of 68%, 27%, and 17%, respectively. There was a significant difference in survival between patients with cirrhosis and those without cirrhosis (P = 0.027). Univariate analysis showed that cirrhosis, vascular invasion, hepatic duct invasion, lymph node metastasis, positive surgical margin (R1), and TNM stage were significantly associated with poor survival. Multivariate analysis showed that cirrhosis, positive surgical margin, and lymph node metastases were related to survival, with hazard ratios of 2.49, 3.53, and 4.16, respectively. CONCLUSIONS: Cirrhosis is an independent factor for poor prognosis in intrahepatic cholangiocarcinoma after surgery.

[Construction and identification of shRNA expressing vectors targeting RhoA gene].

AIM: To construct and identify shRNA interference vector targeting RhoA gene. METHODS: Two pairs of single stranded oligonucleotides encoding RhoA siRNA sequence were designed and synthesized. After annealing, its were inserted into vector pGPU6/GFP/Neo, constructed recombinant vectors and then were identified by restrictive digestion and DNA sequencing. LoVo cells were transfected with the recombinant DNA samples by the liposome complex method, and then fluorescence photographs were taken. RESULTS: Enzyme digestion and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pGPU6/GFP/Neo vector. CONCLUSION: The RhoA gene-targeted siRNA and its vector are successfully constructed.

[Ex vitro sentinel lymph node mapping in colorectal carcinoma].

OBJECTIVE: To investigate the feasibility of ex vitro sentinel lymph node (SLN) mapping with methylene blue staining and its clinical value of predicting regional lymph node metastasis in colorectal cancer. METHODS: Methylene blue (1 ml) was injected submucosally around the tumor immediately after resection. After 2-5 minutes, the first blue-dyed lymph nodes, sentinel lymph nodes (SLNs), were harvested for pathological examination, and compared with the pathological results of other lymph nodes. RESULTS: Of the total 32 patients, 57 SLNs were successfully identified in 30 patients with an average of 1.9 nodes per person. The successful labeling rate was 93.8% (30/32). Among the 13 patients with positive SLNs, there were 5 patients with positive non-SLNs and 8 patients with negative Non-SLNs. Among the 17 patients with negative SLNs, there were 15 patients with negative non-SLNs and 2 patients with positive Non-SLNs. The accuracy of SLNs for regional lymph node metastasis was 93.3% (28/30), the false negative rate was 11.8% (2/17), and the specificity was 100% (13/13). CONCLUSIONS: Ex vitro sentinel lymph node mapping with methylene blue staining in colorectal carcinoma is technically feasible and can effectively reflect the metastatic situation of regional lymph nodes.

[Comparative study on long-term results of laparoscopic and open radical resection for colorectal carcinoma].

OBJECTIVE: To compare the long-term results of laparoscopic and open radical resection for colorectal carcinoma. METHODS: Two hundred and fifteen patients with colorectal cancer from January 1996 to September 2000 were non-randomly divided into laparoscopic and open operation groups. Local recurrence, distant metastasis, 5-year survival rate and long-term postoperative complications were compared between the two groups. RESULTS: Eighty-seven cases received laparoscopic resection and 128 cases received open operation. There were no statistical differences in age, sex and tumor stage between the two groups (P > 0.05). The 5-year-survival rate was 70% in open operation group, and 78% in laparoscopic group (P > 0.05). There were no significant differences in the incidences of local recurrence, distant metastasis, incision seeding, and incision hernia between the two groups (P > 0.05). The complication rate of postoperative adhesive intestinal obstruction was significantly lower in laparoscopic group than that in open operation group (P< 0.05). CONCLUSIONS: Long-term results of laparoscopic resection are similar to those of open resection for colorectal carcinoma, but laparoscopic surgery has less long-term complications.