Objective To screen and identify differentially expressed genes in colorectal carcinoma and explore the possible molecular pathogenesis of colorectal carcinoma.Methods The differentially expressed cDNA bands in colorectal carcinoma specimens and matched adjacent normal tissues were isolated by fluorescent mRNA differential display.Following differential display PCR (DD-PCR),all cDNA fragments were sequenced.By using BLAST software,the sequencing results were compared with Genebank database for homologue analysis.RT-PCR was used to detect the expression of one of the differentially expressed genes in colorectal carcinoma samples were identified by semi-RT-PCR.Results BLAST analysis revealed that the cDNA band was homologous to DDX32 (99%) and its up-regulated expression in colorectal carcinoma tissues was confirmed by RT-PCR (P

OBJECTIVE To investigate the pathogenic change and bacterial antibiotic resistance in lower respiratory tract infection in inpatients of chronic pulmonary heavt disease(CPHD) for recently 3 years and to provide reference for clinical treatment.METHODS Retrospective analysis was conducted on sputum cultivation from CPHD patients who were hospitalized from Jan 2004 to Dec 2006.RESULTS A total of 772 pathogenic strains were isolated,60.1 % of which were Gram-negative bacilli.Pseudomonas aeruginosa,Klebsiella Pneumoniae,Escherichia coli and Acinetobacter baumannii were the main Gram-negative pathogens.Gram-positive bacilli accounted for 13.7%,most of which were Streptococcus pneumoniae,Staphylococcus aureus and so on.Fungi accounted for 26.2%.Imipenem/cilastatin sodium was the most sensitive drug for P.aeruginosa,Acinebacter and Enterobacteriaceae.And vancomycin hydrochloride was the most for S.aureus.Their multiple drug-resistance to anti-microbial agents was serious.CONCLUSIONS Gram-negative bacteria are the majority of the pathogens from CPHD of lower respiratory tract infection in hospital.The pathogens show multiple drug-resistance in drug sensitive test.It is suggested that there be urgent need for surveillance of bacterial resistance and rational use of anti-microbial agents during the clinical therapy.

Objective To investigate the effect of human hepatocellular carcinoma HepG2 cell-derived Exosome on the differentiation of mesenchymal stem cells(MSC)into cancer-associated myofibroblasts(CAF)and the impacts of CAF on liver cancer cell proliferation,migration,and invasion. Methods The protein expression of HepG2 cell-derived Exosome was detected by Western blotting. MSCs were separated from human adipose tissue and cultured with HepG2 cell-derived Exosome(100 ng/nl)to initiate differentiation. The expressions of mesenchymal markers and several interleukins were also detected by Western blotting. HepG2 cells were co-cultured with the conditioned media(CM),in which HepG2 Exosome induced the differentiation of MSC into CAF. The expressions of epithelial and mesenchymal markers were detected by real-time polymerase chain reaction(PCR)and Western blotting. Cell proliferation was assessed using MTS assay. Transwell chambers were used in the in vitro migration and invasion assay. Results HepG2 cell-derived particles expressed CD63,70 kilodalton heat shock proteins,and 90 kilodalton heat shock proteins. With the treatment of HepG2 cell-derived Exosome,the expressions of mesenchymal marker α-smooth muscle actin,fibroblast activation protein α,interleukin(IL)-6,IL-8,and IL-1β were up-regulated,while vascular endothelial growth factor had no significant change. The conditioned media which HepG2 Exosome induced MSC differentiation CAF(CAF-CM)could significantly promote HepG2 cells proliferation(1.075±0.104),compared to BSA control(0.874±0.066,P=0.023)and MSC-CM(0.649±0.034,P=0.0005). CAF-CM could significantly enhance cell migration [(42.5±9.1) cells vs.(18.5±3.1) cells,P=0.001] and invasion [(29.0±3.5) cells vs.(13.1±3.7) cells,P=0.009] compared to its control group. Moreover the conditioned medium which HepG2 Exosome induced MSC to differentiate into CAF could also promote the expressions of mesenchyme-related genes Smad interacting protein 1(P=0.040),β-catenin(P=0.038),fibronectin(P=0.029),and Vimentin(P=0.013)and inhibit the expression of epithelial related genes zonula ocdudens-1(P=0.010).Conclusions Exosome extracted from HepG2 cells can induce human adipose-derived MSC to differentiate into cancer-associated myofibroblasts. CAF-like cells can promote the migration of the liver cancer cell line HepG2.

AIM To explore the effects and mechanism of cobra venom serum on the proliferation in HL60 cells. METHODS Established the HL60 cells as a target to study the growth feature by the action of cobra venom serum.The agarose gel electrophoresis and flow cytometry analysis were used to demonstrate apoptosis. RESULTS Compared with the control group, the cells were inhibited significantly by the action of cobra venom serum.A characteristic DNA "ladder" was detected by using agarose gel electrophoresis. By flow cytometry analysis,it was proved that most apoptosis of HL60 cells occurred when cultured with cobra venom serum. CONCLUSION Cobra venom serum inhibited the HL60 cells in vitro , which was related to apoptosis. This may introduce a new way to the treatment of leukemia.

Objective To observe the clinical efficacy of TCM periodic therapy combined with estradiol progesterone tablets (Diane-35) in the treatment for polycystic ovary syndrome (PCOS), and its effects on serum sex hormone, glucose and lipid metabolism.Methods Totally 74 patients with PCOS were randomly divided into Western medicine (WM) group and Chinese and Western medicine (CWM) group, 37 cases in each group. WM group was given Diane-35, while CWM group was treated with TCM periodic therapy additionally, for three courses, 28 d of one course. Clinical symptoms, sex hormones and glucose and lipid metabolism of two groups were observed before and after treatment and withdrawal treatment for 3 months.Results Two patients in each group were lost to follow-up. Markedly effective rate and total efficiency rate of CWM group were better than WM group (P<0.05). After treatment, symptom integrals of CWM group decreased significantly (P<0.01). Serum E2 increased (P<0.01), while LH, T and LH/FSH decreased in two groups (P<0.01). After treatment and withdrawal treatment for 3 months, improvement of clinical symptoms and sex hormone levels in CWM group was superior to CM group (P<0.01), FPG, FINS, TG, TC and BMI of CWM group decreased in CWM group (P<0.05), and lower than that of CM group (P<0.05,P<0.01). Adverse reaction rates of WM group and CWM group were 17.1% and 5.71% (P<0.05).Conclusion Efficacy of TCM periodic therapy combined with Diane-35 in treatment of PCOS is significant, which can obviously improve patients’ clinical symptoms, and regulate hormone and lipid metabolism disorders, with fewer adverse reactions.

Objective To evaluatethe curative effect of the integrative treatment of point injection, warm acupuncture and Chinese medicine enema for the chronic pelvic inflammatory disease.Methods A total of 86 patients with chronic pelvic inflammatory diseasewere divided into control group and treatment group, 43 patients in each group , according to the random number table. The control group was treated by warm acupuncture combined with Chinese medicine enema, while the treatment group was treated with point injection plus the basis of control group. The levels of serum CRP, IL-1 and IL-6 were detected by enzyme linked immunosorbent assay,the plasma viscosity and hematocrit were detected by automatic blood rheological test instrument,the symptom scores of 2 groups were compared before and after treatment, the clinical efficacy was evaluated and the recurrence rate was observed. Results The clinical effective rate of treatment group patients was 97.7% (42/43), and the control group 81.4% (35/43). The difference between two groups was significant (χ2=6.081,P=0.014). After treatment, the levels of CRP (7.53 ± 3.44 mg/L vs. 10.11 ± 3.02 mg/L,t=-3.696), IL-1β (26.37 ± 13.98 pg/mL vs. 36.33 ± 4.02 pg/mL,t=-4.490) andIL-6 (23.31 ± 10.11 pg/mlvs. 29.56 ± 4.27 pg/ml,t=-3.734)in the treatment group were significant lower than those in control group (P<0.05). The scores of pain (2.13 ± 0.55vs.2.71 ± 0.62,t=-4.589), tiredness (1.07 ± 0.98 vs. 2.53 ± 0.52,t=-8.630), Body cold (1.51 ± 0.51 vs. 2.21 ± 0.67,t=-5.451), menstrual symptoms (1.27 ± 0.97 vs. 2.29 ± 0.78, t=-5.374) and total points (6.13 ± 3.94vs. 8.55 ± 1.82,t=-3.656) in the treatment group were significant lower than those in control group (P<0.05). The plasma viscosity (1.13 ± 0.25 mPa?svs. 1.41 ± 0.32 mPa?s,t=-4.521) and the red blood cells deposited (0.27% ± 0.08% vs. 0.41% ± 0.07%,t=-8.636) in the treatment group were significant lower than those in control group (P<0.05). The recurrence rate of treatment group was none, while 11.43% (4/35) in control group. Thus, the recurrence rate of treatment group was significantly lower than the control group (χ2=5.063,P=0.024). Conclusions The integrative treatment of point injection, warm acupuncture and Chinese medicine enema can reduce the level of inflammatory factors in patients with chronic pelvic inflammatory disease, improve the blood microcirculation, and reduce the recurrence rate.

Objective To explore the effect of chronic intermittent hypoxia (CIH) on hippocampus neuron and growth associated protein-43 (GAP-43) in rats. Methods 40 adult male Sprague-Dawley rats were randomly divided into control group, CIH 1 week group, CIH 3 weeks group and CIH 5 weeks group with 10 rats in each group. The CIH groups were exposed to intermittent hypoxia in designed cabin 8 hevery day for 1 week, 3 weeks and 5 weeks. The pathological changes of hippocampal neurons were observed by HE staining. The GAP-43 expression of hippocampus was detected by immunohistochemical staining. Results There was degeneration and necrosis of hippocampal neurons in the CIH rats, and it became worse gradually along with the hypoxia. Compared with the control group, the GAP-43 expression in hippocampus increased in CIH rats. The peak of GAP-43 expression appeared in the 1st week (P=0.038) and then it decreased in the 3rd week (P=0.382) and it was closed to the normal level in the 5th week (P=0.860). Conclusion Degeneration and necrosis of hippocampal neurons exacerbated gradually along with the hypoxia, while GAP-43 expression changed inconsistently.

Objective:To study the effects of the conditioned medium (CM) from human lung adenocarcinoma cell line A549 on the viability and apoptosis of human umbilical vein endothelial cells (HUVECs) and the role of PI3K-Akt signaling pathway in the process. Methods:HUVECs were cultured with CM of A549 cells. Cell viability was detected by XTT assay. The morphological changes of HUVECs were analyzed by Hoechst 33258 staining and fluorescence microscopy. The apoptosis was detected by flow cytometry. Expression levels of total Akt and phosphorylated Akt were assessed by Western blotting. PI3K inhibitors wortmannin(WT)and Akt1 siRNA(siAkt1)were used to block PI3K-Akt signaling pathway. The mRNA transcription of Akt subtype was determined by RT-PCR.Results:A549 CM significantly increased cell viability after 24 h treatment (P=0.037) and inhibited apoptosis (P=0.001) of HUVECs. CM time-dependently activated phosphorylation of Akt. Akt was phosphorylated at 15 min after CM treatment and reached the peak at 30 min and then tended to decline. Both WT and siAkt1 blocked the effects of CM. Conclusion:The CM of A549 cells increased the survival and inhibit the apoptosis of HUVECs. Akt1 played a significant role in the process.

Objective To study the diagnostic and prognostic value of procalcitonin (PCT)in patients with sepsis in early stage. Methods In the perspective study, 168 patients enrolled were classified into three groups,ncluding 31 cases in control group,37 cases in SIRS group and 100 cases in general sepsis group. The latter were com-posed of moderate sepsis sub-group with 36 cases, severe sepsis sub-group with 40 cases and septic shock sub-group with 24 cases. Indexes of inflammation, SOFA and concentration of PCT in all patients were determined and their cor-relation with sepsis prognosis was analyzed. Results The level of PCT and CRP is 3.1 ± 2.3 μg/L and 34.7 ± 28.0 mg/L in SIRS group,is 10.8 ± 8.1 μg/L and 106.8 ± 69.3 mg/L in general sepsis group respectively. Which are higher than the level of PCT (0.3 ± 0.2 μg/L )and CRP (4.1 ± 2.9 mg/L)in control group (P < 0.01 ). Higher con-centration of PCT and CRP were found in general sepsis groups than these in SIRS group. The difference is prominem.Moreover,an increasing trend of PCT with the more serious of illness was found in subgroup analysis ,but not in CRP.According to the receiver operating characteristic curves( ROC curves) ,The best cutoff values in the diagnosis of sep-sis were >4.395 μg/L for PCT,>51.8 mg/L for CRP and >4.0 for SOFA score. Condusion PCT and CRP are useful diagnostic parameters with high specificity in early sepsis. PCT combining with SOFA score can be used as ide-al quantitative index to estimate the severity of sepsis and prognosis in patients with sepsis.

Objective To evaluate the role of extracellular signal-regulated kinase (ERK) signaling pathway in inhibition of oxygen-glucose deprivation and restoration (OGD/R)-induced apoptosis in rat cortical neurons by sevoflurane and the relationship with mitochondrial permeability transition pore (mPTP).Methods The rat cortical neurons were cultured in vitro and seeded in 6-well or 12-well culture plates.The neurons were randomly divided into 4 groups (n =18 each) using a random number table:control group (group C);OGD/R group (group O);sevoflurane group (group OS);sevoflurane + ERK1/2 inhibitor PD98059 group (group OSP).The neurons were subjected to O2-glucose deprivation for 90 min followed by restoration of O2-glucose supply for 24 h in group O.The neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OS.OGD/R was performed at 1 h after ERK1/2 inhibitor PD98059 30 μmol/L was added,and the neurons were incubated with 2％ sevoflurane for 2 h after OGD/R in group OSP.At 24 h of restoration of O2-glucose supply,the expression of phosphorylated ERK1/2 (p-ERK1/2) in neurons was measured by Western blot,the neuronal apoptosis was detected using Annexin V-FITC/PI double staining combined with flow cytometry,and the opening of mPTP was determined through measuring the optical density at 540 nm.The apoptosis rate was calculated.Results Compared with group C,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly increased in group O (P＜0.05).Compared with group O,the expression of p-ERK1/2 in neurons was significantly up-regulated,and the apoptosis rate and mPTP opening were significantly decreased in group OS (P＜0.05).Compared with group OS,the expression of p-ERK1/2 in neurons was significantly down-regulated,and the apoptosis rate and mPTP opening were significantly increased in group OSP (P＜0.05).Conclusion The mechanism by which sevoflurane inhibits OGD/R-induced apoptosis in rat cortical neurons is related to inhibition of mPTP opening after activation of ERK signaling pathway.