Mechanism of PP242 inducing autophagy and apoptosis in cholangiocytes of polycystic kidney rats / 中国药理学通报

Aim To investigate the molecular mecha-nism of mTORC1/2 inhibitor PP242, which inhibiting cholangiocyte cell preliferation and cystic diliatation via inducing apoptosis and autophagy in the polycystic kid-ney ( PCK ) rats. Methods The expression of p-mTOR and p-Akt in the bile duct epithelial cells was examined by immunohistochemistry. The inhibiting effect of rapamycin and PP242 on cell proliferation ac-tivity on bile duct epithelial cells, the effect of gene si-lence on LC3, Beclin-1 and the effect of the authoph-agy-specific inhibitor 3-methyladenine (3-MA) on cell proliferation were respectively analyzed by WST-1 as-say. The expression of PI3K/Akt signaling pathway re-lated proteins, autophagy-related proteins LC3, Bec-lin-1 and clevead caspase-3, which were treated by PP242 were determined by Western blot. The effect of PP242 on apoptosis was detected by Annexin V/PI double staining and ELISA. The expression of LC3 in cytoplasm was detected by immunofluorescence. The a-bility of rat bile duct epithelial cells spheroid formation was detected by 3D cell culture method, and the cells were treated by single applied with rapamycin and ap- plied rapamycin combined with Rictor gene silencing respectively. Results The protein levels of p-Akt and p-mTOR markedly increased in the bile duct epitheli-um of PCK rats. PP242 inhibited the proliferation of bile duct epithelial cells more effectively than rapamy-cin and showed a dose-and time-dependent manner ( P<0.05 ) . PP242 significantly reduced the levels of PI3K/Akt signaling pathway-related proteins in PCK rat cholangiocytes. PP242 induced apoptosis and auto-phagy, up-regulated the levels of cleaved caspase-3, Beclin-1 and increased the ratio of LC3-II/LC3-I. The combination of Rictor gene silencing and rapamycin was more effective than rapamycin alone in inhibiting cholangiocytes in PCK rats. The inhibitory effect of PP242 on the cell viability was significantly weakened by treatment with 3-MA and knockdown of LC3 and Beclin-1 ( P <0.05 ) . Conclusions PP242 inhibits the proliferation of PCK rat cholangiocytes through PI3K/Akt/mTOR signaling pathway, and the mecha-nism is closely related with autophagy and apoptosis.

Clinicopathological significance of the expression of carbonic anhydrase II in human pancreatic invasive ductal cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To study the clinicopathological significance of the expression of carbonic anhydrase (CA)II protein and mRNA in primary invasive ductal cancer (IDC) of human pancreas.</p><p><b>METHODS</b>The expression of CAII protein in 33 paired paraffin embedded IDC specimens of the pancreas and paired adjacent non-cancerous pancreatic tissues was detected by immunohistochemistry. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to examine the expression of CAII protein and mRNA level in 12 paired fresh IDC specimens of the pancreas and adjuvant non-cancerous pancreatic tissues. The relationship between the protein expression and clinicopathological features was analyzed.</p><p><b>RESULTS</b>Overexpression of CAII protein was shown in 11 cases of pancreatic IDC tissues (33.3%, 11/33), which was much lower than that in paired non-cancerous pancreatic tissues (72.7%, t = 6.275, P = 0.000). The expression of CAII protein had no correlation with tumor position (χ² = 0.992, P = 0.319), differentiation (χ² = 0.866, P = 0.352), TNM stage (χ² = 1.210, P = 0.271) and Lymph node metastasis (χ² = 0.798, P = 0.372), but had bordering statistic sig with the prognosis of the patients (χ² = 3.233, P = 0.072). The median survival time in the patients with high expression of CAII protein was 540 days, while that in the patients with low expression was 320 days. The expression of CAII protein and mRNA was lower in IDC than that in paired non-cancerous pancreatic tissues detected by Western blot and RT-PCR respectively (t = 3.399, P = 0.006; t = 2.281, P = 0.043).</p><p><b>CONCLUSION</b>CAII is down regulated in pancreatic IDC and might be relative with the prognosis.</p>

Case-control study on therapeutic effects of Chinese herb of promoting blood circulation to dissipate blood stasis in the treatment of mudslide injuries / 中国骨伤

<p><b>OBJECTIVE</b>To observe the effects of Chinese herb of promoting blood circulation to dissipate blood stasis on the levels of tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6) in serum of patients with mudslide injuries, and investigate the mechanisms of Chinese herb of promoting blood circulation to dissipate blood stasis in the treatment of mudslide injuries.</p><p><b>METHODS</b>Patients with mudslide injuries who were translated from Zhouqu and Chengxian to Lanzhou from 12th August 2010 were divided into two groups (group A and group B). Twenty-five patients in group A, containing 15 males and 10 females, with an average age of (39.0 +/- 3.9) years. According to AIS scoring system, 1 point in 2 cases, 2 points in 3, 3 points in 17 and 4 points in 3. No patients got 5 points. Based on ISS rating system, 16 cases got 16 points or less, 7 cases rated between 16 and 25, only 2 cases were equal to 25 points or more. Another 25 patients were in group B, including 11 males and 14 females, with a mean age of (40.1 +/- 3.6) years; AIS score showed 3 cases got 1 point, 4 got 2 points, 16 got 3 and 2 got 4 points, no patients got 5 points. Patients in group B were divided into three parts by ISS score:15 cases (16 points or less); 9 cases (range 16 to 25 points) and 1 case (25 points or more). All the patients accepted general physical checkup. Eight patients were treated by surgical treatment in group A (3 patients were treated with open reduction plate fixation, 4 patients were treated with debridement BHID, and 1 patient underwent foreign body removal) and 6 cases in group B (1 patient was treated with craniocerebral surgery,2 patients were treated with chest surgery, and 3 patients were treated with soft tissue debridement). All the patients of two groups were treated by support treatment, detumescence treatment and preventing infection, complications. A seven days course of treatment with detumescence analgesic mixture 50 ml p.o. bid, traumatologic cleansing liquid 20 ml ad us, ext 20 to 30 min qid to group A,continuous treatment of two courses. Using immunometric assay to determine serum levels of inflammatory cytokine TNF-alpha, IL-6 at the 2nd, 7th, 14th days after admission. Analysis of data were done with the help of SPSS 16.0 statistic software.</p><p><b>RESULTS</b>There were no statistical differences of TNF-alpha, IL-6 between two groups at the second day after admission, and there were statistical differences at the 7th and 14th days between two groups. There were significant differences of TNF-alpha and IL-6 levels between the 7th day and the 2nd day, the 14th day and 2nd day after admission.</p><p><b>CONCLUSION</b>The Chinese herb of promoting blood circulation to dissipate blood stasis can inhibit the release of inflammatory factor after traumatism.</p>

Clinical study on the prevention of heterotopic ossification after total hip arthroplasty by Xiaozhong Zhitong / 中国骨伤

<p><b>OBJECTIVE</b>To explore the therapeutic effects of Xiaozhong Zhitong mixture preventing heterotopic ossification (HO) after total hip arthroplasty.</p><p><b>METHODS</b>From July 2006 to October 2009, 154 patients underwent total hip replacement surgery were randomly divided into sham group (group A, 50 cases), indomethacin group (group B, 55 cases) and Xiaozhong Zhitong mixture group (group C, 49 cases). Among 154 patients, 9 cases were primary osteoarthris, 34 cases osteoarthritis secondary to acetabular dysplasia, 98 cases osteoarthritis secondary to avascular necrosis of the femoral head, 2 cases rheumatoid arthritis, 5 cases femoral neck fracture, 6 cases other diseases. Modified Gibson approach was used during the operation. After operation, group A was no preventing treatment, group B was treated by indomethacin 50 mg every time, twice a day; group C was treated by Xiaozhong Zhitong mixture 50 ml every time, twice a day for 4 weeks. Eighteen months after operation was study termination point and X-ray (including the double hip anteroposterior,obturator oblique and iliac oblique film) was used to observe whether heterotopic ossification was formed (Brooker classification was used to evaluate ossification degree); Harris scoring was used to evaluate the function of hip joint,including PAHSS 80 scores and IAHSS 20 scores.</p><p><b>RESULTS</b>All the patients were followed up,with the average of duration of 21.2 months. The condition of heterotopic ossification: for group A,there were 27 cases with heterotopic ossification(54%) ,and Brooker I in 8 cases, II in 9 cases, III in 8 cases and IVin 2 cases; for group B, there were 12 cases heterotopic ossification (21.82%), and Brooker I in 10 cases, II in 2 cases; for group C, there were 11 cases heterotopic ossification(22.45%), and Brooker I in 9 cases, I in 2 cases. There was significant difference among three group in heterotopic ossification by rank test (P<0.05), but no difference between group B and C (P>0.05); there were no significant difference among three groups before treatment in Harris, PAHSS and IAHSS by analysis of variance (one-way ANOVA) (P>0.05), and has significant difference at 18 months after treatment (P<0.01). There were significant difference in Harris, PAHSS and IAHSS before and after treatment at 18 months (P<0.01). LSD-t was used to analyzed the scoring of Harris, PAHSS and IAHSS, there was significant difference among group A and group B and group C (P>0.05), but no difference between group B and C (P<0.01). There were gastrointestinal reaction in 5 of group A, 35 in group B and 4 in group C.</p><p><b>CONCLUSION</b>The effect of Xiaozhong Zhitong mixture on the prevention of heterotopic ossification after total hip arthroplasty is similar to indomethacin, but Xiaozhong Zhitong mixture has the advantages of less side effects and easily acceptance by patients.</p>

Analysis of pancreatic leaking-related risk factors after pancreaticoduodenectomy / 中华外科杂志

<p><b>OBJECTIVE</b>To analysis the risk factors of pancreatic fistula after pancreaticoduodenectomy (PD).</p><p><b>METHODS</b>A retrospective clinical study had been done in 97 patients who underwent PD between June 2001 and June 2006. The two groups were first compared by the univariate analysis;logistic regression was then used to determine the effect of multiple factors on pancreatic fistula. A P-value of less than 0.05 was considered to be statistically significant.</p><p><b>RESULTS</b>Of the 97 patients, 13 patients were identified as having pancreatic fistula. Factors significantly increasing the risk of pancreatic fistula by univariate analysis included preoperative serum total bilirubin (P = 0.038), operative time (P = 0.003) and whether or not Braun anastomosis (P = 0.034), and prophylactic use of somatostatin (P = 0.003) after operation. A multivariate logistic regression analysis revealed the factors most highly associated with pancreatic fistula to be preoperative serum total bilirubin (OR = 11.687, P = 0.021) and postoperative prophylactic use of somatostatin (OR = 0.056, P = 0.020).</p><p><b>CONCLUSIONS</b>Preoperative serum total bilirubin more than 170 mmol/L was a risk factor of pancreatic fistula after PD, and postoperative prophylactic use of somatostatin was a protect factor of pancreatic fistula after PD.</p>

Epidermal growth factor-mediated NF-kappaB activation promotes uPA expression and invasiveness in pancreatic cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To determine the effect of EGF on the invasiveness of pancreatic cancer cells and its related regulatory mechanism.</p><p><b>METHODS</b>The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay. The expression of uPA was assayed by Western blot and RT-PCR. The activity of NF-kappaB was examined by EMSA.</p><p><b>RESULTS</b>EGF significantly increased the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of uPA at both mRNA and protein levels. Furthermore, EGF stimulated the NF-kappaB binding activity, and pretreatment of cells with a NF-kappaB inhibitor, pyrrolidine dithiocarbamate, markedly attenuated EGF-induced NF-kappaB activation. Subsequently, the EGF-induced uPA expression and invasiveness were also inhibited by NF-kappaB inhibitor.</p><p><b>CONCLUSION</b>Our findings indicated that NF-kappaB-mediated up-regulation of uPA expression is responsible for EGF-induced invasiveness in pancreatic cancer cells, and implicate that such anti-NF-kappaB therapy with NF-kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer.</p>

Role of Epidermal Growth Factor in Invasion and Metastasis of Pancreatic Cancer Cells / 中国普外基础与临床杂志

Objective Overexpressions of epidermal growth factor(EGF)and EGF receptor have been associ- ated with progression and invasive phenotype of pancreatic cancer.However,the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood.In this study,effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated.Methods The effects of EGF on the proliferation,adhesion and invasion of pancreatic cancer cells were detected by WST-1 prolif- eration assay,adhesion assay and invasive assay,respectively.The activity and expression of MMP-2 and MMP-9 were examined by zymography,Western blot and RT-PCR,respectively.The activity of NF-?B was examined by EMSA.Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion.The expressions of NF-?B and MMP-9 were significantly increased by EGF,but EGF did not affect the activity and expression of MMP-2.Furthermore,EGF stimulated the NF-?B binding activity.Pre- treatment with NF-?B inhibitors,pyrrolidine dithiocarbamate(PDTC),could significantly inhibit the activity of NF-?B induced by EGF.Meanwhile,the EGF-induced expression and activity of MMP-9,as well as cell invasiveness were also inhibited by NF-?B inhibitor.Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF-?B in pancreatic cancer cells,which implies that NF-?B inhibitant,such as PDTC,may diminish the invasiveness of pancreatic cancer cells.