Volume 89  ·   Issue 34  ·  2009

In recent years, the level of diagnosis and treatment of pancreatic cancer in China has made great progress, especially the surgical treatment has made a new breakthrough. Compared with before 1998, the 5-year survival rate of pancreatic cancer patients has increased from less than 5% to 10%.

Objective To evaluate whether antisense oligonucleotides (ASODN) targeting hTERT mRNA could sensitize human pancreatic cancer cell to gemcitabine in vitro. Methods hTERT mRNA expression was measured by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) assay. Telomerase activity was examined by TRAP, polyacrylamide gel electrophoresis and silver staining. The proliferation capacity and the apoptosis of cancer cells were evaluated by MTT assay and double staining with both Annexin V-FITC and PI. Results Transient transfection in human pancreatic cancer cell with hTERT ASODN diminished the abundance of hTERT mRNA in a concentration-dependent fashion. And 0.2 μmoL/L ASODN decreased the telomerase activity by 0.47-fold in cancer cell. The IC50 value of gemcitabine in ASODN-transfected cell was (0.8±0.2) μmoI/L while that in oligofectamine-transfected control cell (7.3±0.9) μmol/L with a statistically significant difference, hTERT ASODN significantlly increased the gemcitabine-induced apoptosis rate in pancreatic cancer cell. The gemcitabine-induced apoptosis rate in ASODN-transfected cell was 60.28% while that in oligofectamine-transfected control cell 17.34%. Conclusion hTERT antisense oligodeoxynucleotide can increase the sensitivity of pancreatic cancer cell to gemcitabine. The mechanism may be due to the down-regulated expression of hTERT mRNA and telomerase activity.

Objective To investigate the expression of vascular endothelial growth factor C (VEGF-C) and the effect of antisense oligonucleotide (ASODN) upon lymph node metastasis of pancreatic cancer cell. Methods We selected 15 cases of human pancreatic cancer and detected the expression of VEGF-C in primary tumor and lymph node metastasis tissues with immunohistochemistry. Meanwhile, the spontaneous lymphatic metastasis model in nude mice was established with orthotopic implantation for the human pancreatic cancer cell line PANC-1, isolation and culture of primary tumor and spontaneous lymphatic metastasis. The effect of VEGF-C special ASODN upon the apoptosis of pancreatic cancer cell derived from primary tumor and spontaneous lymphatic metastasis were detected by reverse transcription polymerase chain reaction (RT-PCR), enzyme linked immunosorbent assay (ELISA), flow eytometer and terminal deoxytnucleotidyl transferase mediated-dUTP nick end labeling (TUNEL). Results In tissues of human pancreatic cancer, the values of VEGF-C on lymph nodes metastasis were more higher than primary tumor (P<0.05). About the expression of VEGF-C on pancreatic cancer cell derived from spontaneous lymphatic metastasis and primary tumor in nude mice model, the mRNA levels of VEGF-C were 0.87±0.11 and 0.61±0.15 respectively, the VEGF-C levels in culture supernatants were (1682±157) pg/ml and (1404± 128) pg/ml. The expression of VEGF-C on pancreatic cancer cells derived from lymphatic metastasis were also more higher than primary tumor (P<0.05). In vitro and vivo, transfection of VEGF-C ASODN decreased the expression of VEGF-C in pancreatic cancer cell. In control group, scramble-sense oligonucleotide (SODN) group and ASODN group, the apoptosis rates of pancreatic cancer cells derived from lymph node metastasis were (2.8±1.0)%, (5.0±2.1)%, (13.2±2.2)% respectively in vitro, and were (1.8±0.5)%, (2.0±0.7) %, (4.4±1.0) % respectively in vivo, the apoptosis was increased significantly after transfeetion of VEGF-C ASODN (all P<0.01). But pancreatic cancer cells derived from primary tumor were not effected (all P>0.05). Conclusion In human pancreatic cancer and nude mice model, the expression of VEGF-C on lymphatic metastasis was higher than primary tumor. The apoptosis of pancreatic cancer cells derived from spontaneous lymphatic metastasis were promoted by transfection of VEGF-C ASODN specially.

Objective To analyze the value of combined therapy in pancreatic cancer with a poor prognosis. Methods The prognostic factors of pancreatic cancer with different clinicopathological chacteristics and treatment modalities were analyzed retrospectively and the cumulative curve was plotted using the Kaplan-Meier method. Results (1) From January 2001 to December 2005, 233 patients with pathological and cytological diagnosis of pancreatic cancer were analyzed. The median survival of all patients was 8.67 months, the l-year survival rate 29.6% and the 5-year survival rate 4.5%. (2) Weightloss cases had a median survival of 7.7 months versus 10. 0 months for cases without weightloss (P=0.003). Back pain cases had a median survival of 6.5 months versus 9.0 months for cases without back pain (P= 0.015). Cases with normal CA19-9 levels (≤37 U/ml) had a median survival of 11.0 months versus 8.0 months for cases with CA19-9 > 37 U/ml levels (P=0.000). Stages Ⅲ and Ⅳ disease cases had a median survival of 8.7 and 6.3 months versus 16.0 months for cases of Stages Ⅰ+Ⅱ (P=0.000). In the present study, patients suffering from weightloss or back pain or whose CA19-9 was above 37 U/ml or whose TNM stage was Ⅲ/Ⅳ were defined as cases with a poor prognosis. (3) Patients were grouped as pancreatectomy group, cancer-directed treatment goup (including intraoperative iodine-125 seed interstitial brachytherapy, 5-FU interstitial chemotherapy, radiotherapy, chemotherapy and transcatheter arterial infusion chemotherapy) and no cancer-directed treatment group (including cases receiving biopsies or bypass). Median survival of three groups were 14.0, 8.3 and 6.6 months respectively. And the 1-year survival rates were 53.5%, 22.5% and 11.8% respectively while the 5-year survival rate 5.8%, 0 and 0 respectively. (4)Median survival of poor prognostic cases (who suffered from weightloss or back pain or whose CA19-9 was above 37 U/ml or whose TNM stage was Ⅲ/Ⅳ) treated with pancreatectomy were 12.0, 11.0, 12.0 and 7.0 months respectively. Median survival of poor prognostic cases treated with cancer-directed therapy were 7.7, 7.5, 8.6 and 8.0 months respectively. Median survival of poor prognostic cases treated with no cancer-directed therapy were 4.0, 3.0, 4.0 and 4.6 months respectively. Conclusion Optimized combined therapy is recommended for pancreatic cancer patients with poor prognostic factors.

Objective To investigate the longitudinal changes in quality of life (QoL) for gastroesophageal reflux disease (GERD) treated with 52-week rubeprasole over a period of 2-3 years. Methods A multi-center, open-label and randomized 52-week raberprazole trial was conducted in 67 eosinophilic esophagitis (EE) and 31 non-erosive reflux disease (NERD) patients. The follow-up period is 2-3 years after the treatment. Their QoL were evaluated using SF-36 Health Survey Questionnaire and GERD-HRQL scale. The results were compared with those acquired before and after a 52-week proton pump inhibitor (PPI) treatment. Results (1) Both EE and NERD patients improved significantly according to GERD-HRQL scale in scores of reflux symptoms as well as overall satisfaction (12.5 vs 3.5, 20.0 vs 14.0, both P < 0.01) versus the pre-therapy baseline. (2) Both EE and NERD patients had no significant difference in the scale of GERD-HRQL (2.0 vs 3.5, 5.0 vs 4.0, both P >0. 05) and most major domains of SF-36 questionnaire versus the post-therapy baseline(53±17 vs 61±17,t=-2.143, P=0.035). (3) The NERD patients had a higher score of reflux symptoms than the EE patients according to the GERD-HRQL Scale(14.0 vs 3.5, Z=2.377, P=0.017), however there were no significant differences between NERD and EE in 8 major domains of SF-36 questionnaire(P>0.05). Conclusion Long-term and low-dose PPI treatment achieves improvement both in reflux symptoms and QoL in GERD patients and such effects last a long time. At follow-ups, the reflux symptoms of NERD patients are more severe than EE patients. However, the overall QoL has shown little differences between these two subtypes.

Objective To investigate the prevalence and hazard of smoking in Chinese cardiovascular physicians. Methods A total of 4032 cardiovascular-certified physicians were recruited from 386 medical centers in all 31 provinces and municipalities in mainland China between June 1 2008 and August 31 2008. Results The prevalence of smoking in Chinese cardiovascular physicians was 15.2% (95%CI 14.1%-16.3%), 609 (29.8%) (95% CI 27.8%-31.8%) for men was much higher than 3 (0.2%) (95 % CI 0.1%-0.4%) for women (P<0.01). The prevalence of heavy male smoking was 6.9 % (95%CI 5.9%-8.1%)and there was no heavy female smoker. Significant geographic differences were observed among male smokers. Higher smoking rates were seen in the northwest and southwest parts of China. There were significant differences in prevalence of male smoking between areas. The prevalence of history of hypertension, diabetes mellitus and hypercholesterolemia in male smoking group were all significantly higher than in male non-smoking group (all P <0.01) . The 10-year absolute risk and relative risk of coronary heart disease and ischemic cardiovascular disease estimated by Framingham model and Chinese model were all significantly higher in smoking group than in non smoking group (all P<0.01). Conclusion The prevalence of smoking in Chinese cardiovascular physicians is relatively high and smoking cessation should be targeted in this population.

Objective To identify the novel genetic defects in patients with congenital ventricular septal defect (VSD). Methods The clinical data and blood samples from 160 unrelated subjects with congenital VSD were collected and evaluated in comparison with 200 healthy individuals. The coding exons and flanking introns of NKX2-5 gene were amplified by polymerase chain reaction and sequenced using the di-deoxynucleotide chain termination approach. The acquired sequences were aligned with those publicized in GenBank by the aid of program BLAST to identify the sequence variations. The software Clustal W was applied to analyzing the conservation of altered amino acids. Results Two novel heterozygous missense NKX2-5 mutations were identified in 3 of 160 VSD patients. The same triplet substitution of TTC for TCC at codon 179, predicting the conversion of serine into phenylalanine at amino acid residue 179 (Ser179Phe), was detected in two cases. Another transition of CGC into AGC at codon 36, leading to the change of arginine into serine at amino acid residue 36 (Arg36Ser), was detected in a third case. These two mutations were not observed in 200 healthy controls. A cross-species alignment of NKX2-5 encoded protein sequences displayed that the mutated amino acids were highly conserved evolutionarily. Additionally, two single nucleotide polymorphisms including a common c. 63A > G and a rare c. 606G > C were observed. However, the polymorphic frequency distributions in VSD cases were not statistically different from those in healthy controls (c. 63A>G: X2=3.403, P=0.0651; c. 606G>C: X2=3.278, P=0.0702). Conclusion Novel NKX2-5 mutations are identified in patients with ASD. They may provide new insight into the molecular etiology responsible for VSD.

Objective To observe the effects of CYP2E1 enzymatic alteration upon hematotoxicity by benzene poisoning in mice and the interference of alcohol and cureuma longa. Methods Eighty male Kunming mice(KM) were randomly divided into four groups: group A (20 mice injected subcutaneously with sodium chloride 2 ml/kg once every 2 days for 3 weeks as normal control group); group B (20 mice injected subcutaneously with 2 ml/kg benzene once every 2 days for 3 weeks as model group); group C (20 mice drinking freely 10% alcohol per day on the basis of benzene poisoning model as alcohol-treated group) and group D (20 mice given cureuma longa 3 g/kg once a day on the basis of benzene poisoning model as curcuma-treated group). Then the blood samples were collected so as to count the parameters of WBC, Hb, PLT. In addition, the liver cytochrome P450 activity and MN formation rate were measured. Results (1) Compared with group A, WBC, Hb and PLT of group B significantly decreased[ (2.6±1.0)×109/L, (93±11)g/L, (593±81)×109/L]. (2) Compared with group B, the parameters of group C decreased (P<0.05) while the liver cytochrome P450 activity of group C increased. It indicated that alcohol improved the activity of CYP2E1 and the MN formation rate of group C also increased [(8.8±1.8)‰ vs(6.5± 1.0)‰,P < 0.1]. (3) Compared with group B, the parameters of group D improved (P<0.01). The activity of CYP2E1 of group D decreased and the difference was significant (P<0.01). In the meantime, MN formation rate decreased (P<0.01). Conclusions (1) When the liver cytochrome P450 activity were altered, the hematotoxicity of benzene poisoning in mice underwent changes. (2) Alcohol increases the hematotoxicity induced by benzene poisoning in mice. (3) Curcuma longa has preventive effect against the hematotoxity of benzene.

Objective To investigate the change of proliferation and invasiveness of non-small cell lung cancer (NSCLC) cell lines SPC-A-1, A549 and LTEP-a-2 with fockhead box M1 (FoxM1) expression deficiency. Methods A siRNA targeting FoxM1 was designed to deplete the FoxM1 expression of these cell lines and an unrelated siRNA used as control. Real-time RT-PCR and Western blotting were used to examine the FoxM1 expression in mRNA and protein level respectively. Colony assay, wound healing assay and transwell chamber assay were employed to evaluate the colony formation ability and invasiveness of FoxM1 deficient cells. Results The designed siRNA could efficiently deplete FoxM1 expression by 83.9%, 83.6% and 88.6% in SPC-A-1, A549 and LTEP-a-2 cell lines respectively. Real-time RT-PCR and Western blot test showed that the FoxM1 protein was also depleted. The colony formation numbers (136.0± 15.5, 87.0±2.6 and 121.7±9.4 respectively) and invasion cell numbers (19.2±2.5, 4.2±0.8 and 6.2±1.8 respectively) of FoxM1 deficient SPC-A-1, A549 and LTEP-a-2 cell lines were significantly fewer than those of the unrelated-siRNA transfected group (colony formation numbers were 222.3±20.5, 164.7±14.1 and 260.7±13.5 respectively, and invasive cell numbers were 81.4±6.2, 39.2±4.6 and 35.6±3.0 respectively, all P<0.01). The cell migration rate of siFoxM1 deficient SPC-A-1 (52.6%± 7.8%) was significantly lower than that of the unrelated-siRNA transfected group (85.3%±18.6% ,P < 0.01). Conclusions The proliferation and invasiveness of several NSCLC cell lines were significantly inhibited after the FoxM1 gene expression was depleted. It suggests that inhibiting the FoxM1 expression might be a promising way for lung cancer therapy.

Objective To explore the effects of respiratory syncytial virus (RSV) upon the expressions of mRNA and protein of intercellular adhesion molecule-1 (ICAM-1) in human embryonic lung fibroblast cells. Methods The expressions of mRNA and protein of ICAM-1 were determined in human embryonic lung fibroblast cells after being infected by the LONG strain type A RSV and in normal fibroblast cells with real-time PCR and flow cytometry. Results The mRNA of ICAM-1 expression in human embryonic lung fibroblast cells was 2.51 times at 24 h post-infection as that in normal fibroblast cells(P< 0.05). The protein of ICAM-1 expression of RSV control group(1.25±0.09, 1.87±0.18, 4.78±0. 52, 13.34±0.64, 1.58±0.37) were significantly higher than those of normal cell group(0.21±0.06,0.30± 0.06,0.29±0.07,0.35±0.17,0.35±0.14)at 12, 24, 48, 72 and 96 h post-infection(all P<0.01). The protein of ICAM-1 expression of RSV control group achieved a peak value between 48 h and 72 h, and then it decreased significantly at 96 h. Conclusion Lung fibroblast cell and ICAM-1 may play some roles in pathogenic mechanism of RSV viral pneumonia.

Objective To investigate the effects of nuclear factor-κB (NF-κB) siRNA upon dextran sulphate sodium (DSS)-induced colitis. Methods Thirty-six BABL/C mice were randomly divided into 4 groups: normal control group, NF-κB siRNA, scrambled siRNA and DSS group. Colitis was induced by treatment with 5% DSS in drinking water and evaluated by disease activity index (DAI) and histological score. The tumor necrosis factor (TNF-α) level of colonic mucosa was measured by enzyme linked immunosorbent assay (ELISA). NF-κB p65 expression was determined by immunohistochemical staining. Results Compared with the scrambled siRNA (DAI 3.42±0.67, histological score 4.65±1.53) and DSS group(3.73±0.55,4.47±1.52), a significant decrease was observed in DAI (2.31±0.26) and histological score (2.19±0.77)in mice with NF-κB p65 siRNA (both P<0.05). The expression of NF-κB p65 and TNF-α [(266±35) pg/ml] in mice with DSS-induced colitis was significantly reduced after treatment with NF-κB 1065 siRNA (P<0.05) in comparisons with the scrambled siRNA[(412±51)pg/ml] and DSS group pg/ml[(449±44)pg/ml]. Conclusion NF-κB p65siRNA shows protective effects upon the mice with DSS-induced colitis. Blockade of NF-κB pathway by NF-κB 1065 siRNA may serve as a novel gene therapy for ulcerative colitis.

Objective To explore the remodeling mechanism of myocardium and sympathetic nerve in pressure overload left ventricular hypertrophy and elucidate the protective effect of statins. Methods Pressure-overload left ventricular hypertrophy (LVH) of rats was induced by partial coarctation of abdominal aorta; a sham-operated group served as the control (SHAM, n=22). At 8 weeks pest-operation, the animals were divided into two groups and a 12-week treatment period was investigated. At the end of treatment period, eehocardiographic evaluations and hemodynamic measurements were performed. Sympathetic innervation was investigated by analyzing nerve growth factor (NGF), growth associated protein-43 (GAP43) and tyrosine hydroxylase (TH). Results In LVH rats, a significant increase of left ventricular weight, left ventricular weight/body weight, echocardiographic left ventricular end-diastolic diameter, interventricular septum thickness, posterior left ventricular wall thickness, left ventricular systolic pressure and dP/dt was observed. The expressions of NGF and GAP43 protein were significantly down-regulated (0.82±0.06 vs 1.53±0.10, 0.68±0.06 vs 0.81±0.10) and TH level was up-regulated (0.44±0.10 vs 0.62±0.06) by RSV treatment. Conclusion A HMG CoA inhibitor reverses the development of left ventrieular hypertrophy and inhibits sympathetic innervation in abdominal aortic-clamped animals.

Objective To investigate the significance of pathological changes in murine lung by a single intramuscular injection of chemokine-like factor 1 (CKLF1). Methods A total of 120 gender-matched BALB/c mice were randomly and evenly divided into treatment group and control group (60 in each). One hundred nanomilligram of pcDNA3. 1-CKLF1-Myc-His, CKLF1-expressing plasmid, in 100 μl of pyrogen-free saline was injected into the anterior tibial muscle of mice followed by the delivery of electric pulses. Mice in the control group received 100 μg of pcDNA3.1-Myc-His in 100 μl of pyrogen-free saline. At the end of week 1,4 and 8 respectively after injection of CKLF1, 20 mice were sacrificed in every group and the cellular profiles in bronchoalveolar lavage fluid (BALF) and the pulmonary pathological changes were observed. Results At the end of week 1 and 4 respectively after CKLF1 injection, the neutrophils [(35.0±5.2) % and (22.9±2.2)% respectively] and lymphocytes [(34.5±2.8) % and (22.0± 2.0)% respectively] in BALF of the treatment group were higher than those of the control group [neutrophils: (6.7±2.2)% and (7.0±2.4)% respectively, lymphocytes: (5.9±1.6)% and (6.1± 2.7) % respectively, all P < 0.01]. Pathological studies demonstrated shedding of bronchiolar epithelium, congestion and edema in interstial tissue and inflammatory cell infiltration in mice at 1 week after CKLF1 injection. Week 4 after CKLF1 administration, the alveolar wall was shown significantly thickened with proliferation of neutrophils, macrophages and fibroblasts as well as remarked collagen deposition in the interstitium. At the end of week 8 after CKLF1 administration, the remarkable morphological changes of the lung gradually subsided and the structure of the lung returned to normal. Conclusions CKLF1 causes injury of inflammation and remodeling in airway in mice. The pulmonary pathological changes induced by a single intramuscular injection of CKLF are reversible.

Objective To observe the alterations of pulmonary function in infants with respiratory diseases. Methods A total of 900 infants with respiratory diseases were recruited and pulmonary function measured in 30 healthy infants. The tests were performed in the sleeping infants with sedation. Tidal breathing flow-volume (TBFV) loops were recorded when infants were breathing quietly. Passive flow-volume technique was used to obtain static respiratory system compliance and resistance. Functional residual capacity was measured by body plethysmograph. Results The TBFV loop showed proximate round or oval curve in healthy infants. The shape and parameters of TBFV loop had significant differences in infants with respiratory diseases as compared with healthy controls. The TBFV loop displayed a concave expiratory curve and ratio of time to reach tidal peak flow to total expiratory time, the expiratory volume till peak flow divided by the total expiratory volume significantly decreased in infants with small airway obstruction. The expiratory or inspiratory curve showed a plateau and the ratio of mid-expiratory to mid-inspiratory flow was less than 60% or over 150% in infants with upper airway obstruction. The TBFV loop turned narrow and lung volume decreased in infants with restrictive diseases. Conclusion The TBFV loop show proximate round or oval curve in healthy infants. Pulmonary function has significant differences between healthy controls and infants with respiratory diseases. Pulmonary function test is useful in the assessment of respiratory diseases.

Primary pulmonary sarcoma is a malignant tumor derived from lung mesenchymal tissue, accounting for about 0.1% ~3.4% of lung malignant tumors [1-2].

A 71-year-old male was admitted to hospital due to repeated chest tightness and shortness of breath for 9 years and depressed edema of lower limbs for 4 years. Nine years ago, he developed chest tightness and shortness of breath without obvious triggers, no obvious chest pain and pressing sensation in the precordial area, and no banding sensation in the chest and back. The symptoms were aggravated after activity and relieved after rest.

A 93-year-old male was admitted to the hospital due to fever and cough. He weighed 66 kg and had a body mass index of 24.2 kg/m2 at the time of admission. The risk factors were advanced age, poor swallowing function, poor gastrointestinal motility, recurrent chronic pulmonary infection, controlled type 2 diabetes mellitus and adjusted to normal hypothyroidism.

The incidence of invasive fibromatosis (AF) is (2~4) ×10-6, accounting for 0.03% of all tumors and 3% of all soft tissue tumors. The disease was first proposed by MacFarlane in 1832. The prevalence of the disease is 15~60 years old, and the average onset age is 30 years old. There are more women than men [1]. There is no obvious racial difference in the occurrence of the disease.

Negative ventricular remodeling and heart failure caused by massive myocardial cell necrosis and fibrous scar formation after acute myocardial infarction are the main problems affecting long-term prognosis. Because excessive degradation of extracellular matrix (ECM) after myocardial infarction can lead to ventricular remodeling and progressive cardiac insufficiency, it is suggested that a new treatment method for myocardial infarction is to implant biomaterials instead of damaged ECM and improve the local microenvironment in the infarcted area [1-2].

Cardiopulmonary resuscitation (CPR) is a rescue measure for respiratory and circulatory arrest. Artificial respiration is used to replace the patient's spontaneous breathing, and cardiac compression is used to form temporary artificial circulation and induce the spontaneous beating of the heart. The purpose is to quickly establish effective artificial circulation, provide oxygenated blood to brain tissue and other important organs, and make them protected.

In March 2009, the respiratory disease caused by novel swine influenza A (H1N1) virus (S-OIV) broke out in Mexico. As of May 29, there were 4,910 confirmed cases of S-OIV infection and 85 deaths in Mexico.

A 50-year-old male was admitted to the hospital on March 22, 2009 due to "abdominal pain for 3 days". In the past 3 days, he developed dull pain under the xiphoid process without inducement, accompanied by nausea, vomiting and fever. The pain aggravated and scleral yellowing and fatigue appeared.