An experimental study of gemcitabine inducing pancreatic cancer cell apoptosis potentiated by nuclear factor-kappa B P65 siRNA / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism of NF-kappaB P65 gene silencing by small interference RNA on the apoptosis of human pancreatic cancer cells induced by gemcitabine in vitro and in vivo.</p><p><b>METHODS</b>Human pancreatic cancer cells (BxPC-3 and PANC-1) were cultured and respectively divided into five groups: blank control group, negative control siRNA group, gemcitabine group, NF-kappaB P65 siRNA group and gemcitabine + P65 siRNA group. The ability of cell proliferation was analyzed by MTT; the expression of NF-kappaB P65 and the apoptosis related proteins were examined by Western blot assay; the apoptosis was evaluated by the flow cytometry and laser scanning confocal microscopy analysis stained with Annexin V-FITC/PI; the DNA binding activity of NF-kappaB was examined by electrophoretic mobility shift assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors. The tumor volume was monitored and TUNEL assay was used to assess the apoptosis index in tumor tissue after treatment.</p><p><b>RESULTS</b>At 72 h after transfection, the combination with gemcitabine and p65 siRNA significantly decreased the cell viability index (P < 0.05), and down-regulated the expression of Bcl-2 and procaspase-3 and up-regulated the expression of Bax compared with other groups. The combined treatment significantly increased the rate of apoptosis compared with other groups (P < 0.05). EMSA assay indicated that the DNA binding activity of NF-kappaB significantly decreased in NF-kappaB P65 siRNA group and gemcitabine+P65 siRNA group compared with Control group. The combined therapy inhibited the growth of pancreatic xenograft tumors by apoptosis induction in nude mice (P < 0.01).</p><p><b>CONCLUSIONS</b>The effect of gemcitabine inducing cell apoptosis may be potentiated through inhibiting the DNA binding activity of NF-kappaB and regulating the expression of apoptosis related proteins by NF-kappaB P65 siRNA, which can activate the mitochondria apoptosis pathway in pancreatic cancer in vitro and in vivo.</p>

Experimental study of the function and mechanism combining dihydroartemisinin and gemcitabine in treating pancreatic cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor activity of combined gemcitabine with dihydroartemisinin, and the mechanism of the anti-tumor effect of gemcitabine enhanced by dihydroartemisinin on pancreatic cancer.</p><p><b>METHODS</b>For cultured cells, cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis and confocal laser scanning microscope stained with Annexin V-FITC/PI. The nuclear extract for determining NF-kappaB DNA-binding activity was analyzed by EMSA, while nuclear P65 and its downstream gene expression was determined by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to agents. TUNEL assay was used to assess tumor cell apoptosis in tumor tissue.</p><p><b>RESULTS</b>After combination of gemcitabine and dihydroartemisinin treatment, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and Panc-1 reached up to (81.1 +/- 3.9)% and (76.5 +/- 3.3)%, and the apoptosis rates were up to (53.6 +/- 3.8)% and (48.3 +/- 4.3)%, the differences were significantly (P < 0.01) compared with gemcitabine [(24.8 +/- 2.9)% and (21.8 +/- 3.5)%]. All the treatment groups inhibited the growth of pancreatic xenograft tumors in nude mice. The tumor volume and apoptosis index were (262 +/- 37) mm(3) and (50 +/- 4)% respectively in the combined treatment, compared to those of [(384 +/- 56) mm(3) and (25 +/- 3)%] in gemcitabine, the differences were significantly (P < 0.05). EMSA showed that gemcitabine alone obviously enhanced its DNA-binding activity compared to control. However, dihydroartemisinin significantly reduced its DNA-binding activity, so that abrogated the inducing effect of gemcitabine on NF-kappaB activation. Western blot assay indicated that dihydroartemisinin downregulated expression of nuclear P65, and combined treatment not only downregulated the expression of Cyclin D1, Bcl-xL and Bcl-2 while upregulated Bax, thus reduced the Bcl-2/Bax ratio, but also increased the caspase-3 activation, all of which increased apoptosis in both BxPC-3 and Panc-1 cells.</p><p><b>CONCLUSION</b>Dihydroartemisinin significantly abrogated the inducing effect of gemcitabine on NF-kappaB activation and downregulated the expression of NF-kappaB targeted gene products, which may be one possible mechanism by which dihydroartemisinin augments the anti-tumor effect of gemcitabine on pancreatic cancer.</p>

Study on anticancer effect of dihydroartemisinin on pancreatic cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo.</p><p><b>METHODS</b>For cultured cells, cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITC/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue.</p><p><b>RESULTS</b>After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 +/- 3.5)% and (79.5 +/- 2.9)%, and the apoptosis rates were up to (55.5 +/- 3.2)% and (40.0 +/- 3.5)%, the differences were significantly (P < 0.01) compared with control [(2.0 +/- 1.3)% and (0.9 +/- 0.4)%]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptosis index were (49.1 +/- 3.9)% and (50.2 +/- 4.4)% respectively in dihydroartemisinin 50 mg/kg treatment group, compared to those of (72.1 +/- 3.3)% and (9.4 +/- 2.9)% in control, the differences were significantly (P < 0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21(WAF1) and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells.</p><p><b>CONCLUSIONS</b>Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptosis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.</p>

The effect of hyperbaric oxygen on acute pancreatitis through downregulating hypoxia-inducible factor / 中华外科杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of hyperbaric oxygen (HBO) on acute pancreatitis (AP) by downregulating hypoxia-inducible factor (HIF).</p><p><b>METHODS</b>Forty Wistar rats were randomly divided into 4 groups (n = 10): sham group, AP group, normo-oxygen group (NP) and HBO group. At 4 hours after taurocholate-induced AP, the rats of NP group and HBO group were respectively treated with oxygen or HBO for 90 min. Several parameters were measured to evaluate oxygen stress after treatment including oxygen saturation (SaO2), partial pressure of oxygen (PO2), pH, and serum LDH. Pancreatic tissues were subjected to histopathological analysis, immunostained, and homogenized for Western blotted analysis of HIF-1alpha and VEGF, and measuring myeloperoxidase activity. The serum TNF-alpha and pancreatic histopathological scores were evaluated the severity of AP.</p><p><b>RESULTS</b>It was proved by immunohistochemisty that HIF in acinar cell and polymorphonuclear leukocytes (PMNs) was activated and transferred from cytoplasm into nucleus in AP group, NP group, and HBO group, following upregulation of VEGF. HBO therapy elevated blood SaO2 (99.6% +/- 0.7% vs. 87.7% +/- 1.8% or 91.2% +/- 2.5%, P < 0.05) and PaO2 [(369.1 +/- 67.6) mm Hg (1 mm Hg = 0.133 kPa) vs. (86.6 +/- 5.6) mm Hg or (99.9 +/- 4.0) mm Hg, P < 0.05]. HBO therapy attenuated the severity of AP through inhibiting AP-induced upregulation of HIF-1alpha and VEGF, as evidenced by reducing histopathological scores (12.40 +/- 1.21 vs. 16.45 +/- 1.10 or 16.38 +/- 1.10, P < 0.05), dry/wet weight ratio of pancreatic tissues, and myeloperoxidase activity.</p><p><b>CONCLUSIONS</b>HIF-1alpha plays a key role in the pathogenesis of AP. HBO therapy attenuates the severity of AP through downregulating the expression of HIF-1alpha.</p>

Regulatory Effect of Qing Yi Tang on Inflammatory Response in Rats with Acute Pancreatitis and Endotoxemia / 中华医院感染学杂志

OBJECTIVE To study the effect of Qing Yi Tang on acute pancreatitis(AP),especially AP complicated with endotoxemia and its possible mechanism.METHODS Fourty Wistar rats were divided into 5 groups including group A(the control group),group B(the AP group),group C(the AP group treated with Qing Yi Tang),group D(the AP group treated with LPS) and group E(the AP group treated with LPS + Qing Yi Tang).Pathological damage of pancreatic tissue was scored with HE staining.The mRNA expression of TNF-? was measured with semi-quantitative RT-PCR,and activation of NF-?B was detected with flow cytometry(FCM) assay.RESULTS It was shown in results that the expression of TNF-? mRNA,activation of NF-?B and pathological damage of the group B were all obviously higher than those of the group A.After treated with LPS which might promote the activation of NF-?B,there was seen the further rise of the activation of NF-?B,expression of TNF-? mRNA and pathological damage.When Qing Yi Tang intervention was applied,the activation of NF-?B and the expression of TNF-? mRNA could be remarkably relieved,so did the pathological damage of pancreas.CONCLUSIONS Qing Yi Tang may be applied to decrease activation of NF-?B and the expression of TNF-? so as to treat AP or AP with endotoxemia.

Experimental study on electroacupuncture for strengthening gastrointestinal motility in the rat with acute pancreatitis / 中国针灸

<p><b>OBJECTIVE</b>To explore the mechanism of EA in treatment of acute pancreatitis.</p><p><b>METHODS</b>Twenty-four healthy male Wistar rats were randomly divided into 3 groups, a control group, a model group and an EA group. In the model group, rat acute pancreatitis model was prearpared by intraperitoneal injection of Caerulein, and in the EA group, EA was given at "Zusanli" (ST 36) and "Tianshu" (ST 25) of the model rat. The gastric emptying rate, small intestinal impelling ratio, myeloperoxidase activity in the pancreas tissue, pathological score of the pancreas and serum amylase were detected.</p><p><b>RESULTS</b>Compared with the control group, both the gastric emptying rate and the intestinal impelling ratio significantly decreased in the model group (P<0.05), and they significantly increased in the EA group compared with the model group (P<0.05). Myeloperoxidase activity in the pancreas tissue, pathological score of the pancreas and serum amylase activity significantly decreased in the EA group as compared with the model group (P<0.05).</p><p><b>CONCLUSION</b>EA can significantly improve the disturbance of gastrointestinal motility induced by acute pancreatitis and relieve pathological damage of pancreas.</p>

Different cell death modes of pancreatic acinar cells on macrophage activation in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pathogenesis of acute pancreatitis is complex and largely unclear. The aim of this study was to explore the relationship between modes of cell death in pancreatic acinar cells, the release of cell contents and the inflammatory response of macrophages.</p><p><b>METHODS</b>Our experiment included four groups: group A (the control group), group B (AR42J cells overstimulated by caerulein), group C (AR42J cells treated with lipopolysaccharide and caerulein), and group D (AR42J cells treated with octreotide and caerulein). Apoptosis and oncosis, and the release of amylase and lactate dehydrogenase (LDH) from AR42J cells were detected. Rat macrophages were stimulated by 1 ml supernatant of culture medium of AR42J cells. Finally, NF-kappaB activation and TNF-alpha and IL-1beta secretion by macrophages were detected.</p><p><b>RESULTS</b>Oncotic cells in group C increased while apoptotic cells decreased (P < 0.05); cells in group D had the inverse reaction. The release of amylase and LDH changed directly with the occurrence of oncosis. The transcription factor NF-kappaB was activated and secretion of TNF-alpha and IL-1beta were significantly higher in group C than in group B (P < 0.05); in group D, these actions were significantly lower than in group B (P < 0.05). This trend was in line with changes in amylase and LDH production.</p><p><b>CONCLUSION</b>There is a close relationship between modes of pancreatic acinar cell death, the release of cell contents and the inflammatory reaction of macrophages.</p>

Regulating effects of arsenic trioxide on cell death pathways and inflammatory reactions of pancreatic acinar cells in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is accepted that inflammatory cytokines play a key role in the development of acute pancreatitis, so blocking the initiation of inflammatory reactions may alleviate pathological changes of acute pancreatitis. We studied the regulatory effect of arsenic trioxide (As(2)O(3)) on apoptosis and oncosis of pancreatic acinar cells in vitro and in vivo and its therapeutic effect on acute pancreatitis.</p><p><b>METHODS</b>Pancreatic acinar cells were isolated by collagenase digestion method. Apoptosis and oncosis of isolated pancreatic acinar cells were detected with Hoechst 33258 + PI or Annexin V + PI double fluorescent staining. Amylase and lactate dehydrogenase release were measured. Acute pancreatitis was induced in Wistar rats by intraperitoneal injections of caerulein, and apoptosis was detected with terminal dUTP nick-end labeling method. Tumor necorsis factor alpha (TNF-alpha) mRNA, myeloperoxidase, nuclear factor-kappaB and histological grading of pancreatic damage were measured.</p><p><b>RESULTS</b>There was an increased apoptosis but a decreased oncosis of pancreatic acinar cell after the treatment with As(2)O(3). The levels of lactate dehydrogenase and amylase release were markedly decreased in As(2)O(3) treated group. Myeloperoxidase content, TNF-alpha mRNA level, nuclear factor-kappaB activation and pathological score in As(2)O(3) treated group were significantly lower than in the untreated group.</p><p><b>CONCLUSIONS</b>As(2)O(3) can induce apoptosis and reduce oncosis of pancreatic acinar cell, thus resulting in reduced release of endocellular enzyme of acinar cells, reduced inflammatory cell infiltration and decreased the production of inflammatory cytokines, so that the outcome of alleviated pathological changes was finally achieved.</p>

The importance of nonrecurrent laryngeal nerve in thyroid surgery / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the anatomic variation of nonrecurrent laryngeal nerve (NRLN) and its surgical identification and prevention during thyroidectomy.</p><p><b>METHODS</b>The database of 5 NRLN cases was analyzed to investigate the difference of operative maneuvers and procedures.</p><p><b>RESULTS</b>All 5 NRLN were located in the right side. Two cases were found have vocal cord paralysis and 1 case recovered in 3 cases who have NRLN injures.</p><p><b>CONCLUSIONS</b>Any transverse bond should not be cut between vascular and laryngeal except middle thyroid vein. Recurrent laryngeal nerve (RLN) should be dissected during thyroid excision. Cervical pneumogastric nerve should be systematic dissected to detect whether RNLN is exist, if RLN is not exist in the same side.</p>

Effect of electroacupuncture on myoelectric activity of Jejunal limb After Roux-en-Y esophagojejunostomy / 中国针灸

<p><b>OBJECTIVE</b>To explore the effect of electroacupuncture on myoelectric activity of Jejunal limb after Roux-en-Y esophagojejunostomy.</p><p><b>METHODS</b>Fourteen health young pigs were randomly divided into 2 groups, an experimental group (total gastrectomy and Roux-en-Y esophagojejunostomy was carried out) and a control group (the abdominal cavity was closed after the electrode was placed), 7 pigs in each group. Electroacupunture was given at "Zusanli" (ST 36) in the experimental group. The changes of myoelectrogram of the jejunal limb was investigated.</p><p><b>RESULTS</b>Compared with the control group, the amplitude and the frequency of the slow wave, and the amplitude and incidence rate of the spike potential in the experimental group were changed significantly; the duration of migrating motor complex (MMC) phase III was (2.6 +/- 0.7) minutes in the experimental group, which was significantly shorter than (7.1 +/- 1.1) minutes in the control group. Electroacupuncture did not significantly influence the amplitude and the frequency of the slow wave, but could increased significantly the incidence rate and the amplitude of the spike potential; after electroacupuncture, the duration of MMC phase III was (5.7 +/- 0.9) minutes, which was significantly longer than (2.6 +/- 0.7) minutes before electroacupuncture.</p><p><b>CONCLUSION</b>Electroacupuncture at "Zusanli" (ST 36) can relieve the Roux-en-Y stasis syndrome through influencing myoelectric activity of the jejunum.</p>

Surgical treatment of acute obstructing colorectal cancer: a report of 297 cases / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To evaluate surgical treatment of obstructing colorectal cancer.</p><p><b>METHODS</b>From July 1993 to July 2003, clinical data of 297 cases undergoing emergency operation for obstructing colorectal cancer were analyzed retrospectively. There were 103 cases with right-sided lesion and 194 cases with left- sided lesion.</p><p><b>RESULTS</b>All patients received emergency operation. Stage i tumor resection and anastomosis were performed in 126 patients including 98 cases with right- sided lesion and 28 with left- sided lesion, total or subtotal colectomy in 108,Hartmann operation in 36,Dixon operation in 9, ileocolic anastomosis in 11,and colostomy in 7 cases. Postoperative complications occurred in 53 cases (17.8% ) including incision infection, intraperitoneal infection and intestinal fistula. There were 17 perioperative deaths. Two hundred and eighty cases healed (94.3% ).</p><p><b>CONCLUSION</b>Stage i tumor resection and anastomosis and total or subtotal colectomy are feasible and safe surgical procedures for obstructing colorectal cancers.</p>