The INK4a-ARF locus is located on CDKN2A point in human chromosome 9p21 and is known to encode two functionally distinct tumor-suppressor genes p16INK4a (MTS1/CDKN2/INK4a/p16) and p14ARF, have been shown to play significant roles in cell-cycle regulation in cancer. p16 is encoded by the exon 1a-exon 2-exon 3 transcript and functions as a negative regulator of the cell cycle through its inhibition of cyclin-dependent kinases (CDKs) 4 and 6 and subsequent blockage of the cyclindependent phosphorylation of pRB . Thus, loss of p16 function leads to deregulation of pRB's suppressive block of the G1-to-S transition and cell proliferation. In contrast, p14ARF is encoded by the exon 1?-exon 2 transcript and acts upstream of p53. Biochemical evidence has demonstrated that p14ARF physically interacts with human double minute 2 (HDM2) gene product and consequently stabilizes p53, leading to G1 cell-cycle arrest. Overall, somatic mutation of the INK4a/ARF tumor suppressor locus, resulting in functionally deficient p16 and possibly p14ARF proteins, seems to be a prevalent event in the development of oesophagus carcinoma.

Objective To explore the relationship of serum leptin concentration,body mass index (BMI) and esophageal squamous cell carcinoma.Methods Blood samples were collected from 47 patients with esophageal squamous cell carcinoma and 20 healthy controls in Shanxi Cancer Hospital.Serum leptin was measured by ELISA.Results Serum leptin concentration in patients group was significantly higher than control group[(13.09±5.94) ng/ml vs (7.58±4.16) ng/ml,(t =3.76,P =0.001)].In 47 patients,BMI,albumin and lymphocytes were (25.20±3.14) kg/m2,(43.58±2.15) g/L and (2.02±0.55)×109/L,respectively.The serum leptin concentration of all patients (47 cases) were positive correlation with BMI (r =0.561,P =0.006),not with blood albumin and lymphocytes (r =0.206,P =0.242; r =0.122,P =0.412).Multiple logistic regressionanalysis reveal statistically significant association between serum leptin and esophageal squamous cell carcinoma incidence (OR =1.442,95 ％ CI 1.094-1.848,P=0.09).Conclusion Higher serum leptin levels seem to represent an additional and independent risk factor for esophageal squamous cell carcinoma.

Objective To explore the clinical significance and the impact on prognosis of blood micrometastasis in the patients with pN0 esophageal squamous cell carcinoma. Methods Total RNA was extracted with TRIzol and mRNA was transcribed reversely into cDNA. RT-PCR was used to detect MMP-7 mRNA and hTERT mRNA in blood. △△Ct sample values were calculated with post-operative follow-up of 3 month, 6 month, 12 month. Results Statistical results suggested that blood micrometastasis was related to differentiation grade and pTNM staging (P=0.000, P=0.000 respectively), but not to age, sex, length of turnout (P0.05). Follow-up results suggested that the degree of invasion and tumor metastasis (recurrence) was no correlation; follow-up to 6 month and 12 month, tumor metastasis (recurrence) was associated with blood micrometastasis, and follow-up to 12 month, compared with the tumor metastasis (recurrence) probability of blood micrometastasis-positive patients and negative patients, the former was as 6.44 times as the latter. (OR=6.440, 95 % CI 1.547-26.822). Conclusion Blood micrometastasis testing is of great significance to early diagnosis and prognosis judgment in pN0 esophageal squamous cell carcinoma patients.

Objective To investigate the drug sensitivity of cardia tumors in vitro by ATP-TCA, and to provide basis of clinic chemotherapy. Methods 42 cardia tumors were tested by ATP-TCA to determine and assess the drug sensitivity of 8 common chemotherapeutics. Correlation analysis was done between the clinical classification and the drug sensitivity. Results The drug sensitivity had significant difference among cardia tumors. From high to low, it was ADM>MMC>PTX> 5-Fu>HCT =VP16>CDDP>IFO, Sensitive degree were 50.00 %, 35.45 %, 31.25 %, 23.53 %, 10.53 %, 10.53 %, 7.69 %, 5 %. Sensitivity of the same drug in different pathologic stages had no significant difference. Conclusion ATP-TCA can play a guiding role in individual chemotherapy for human cardia cancer.

Objective To study the function of γδT lymphocytes in patients with lung cancer.Methods γδT lymphocytes in peripheral blood of the 124 preoperative patients with lung cancer(experiment group) and 56 healthy donors(control group) were detected by Flow Cytometry. Results The percentage of γδT cells(CD+3 γδTCR+) was (6.54±2.90) % in control group and (3.76±2.81) % in experiment group (t =3.568,P ＜0.001). The γδT cells in peripheral blood of patients with lung cancer was less than that of healthy donors (t =3.568, P ＜0.001). There was significant difference in expression of γδT cells in the peripheral blood between the all test groups and healthy control group (P =0.000). Compared with control group, γδT cells of the peripheral blood in experiment group were correlated with TNM stage, general types, pathological changes position and histology grades (P ＜0.05). The expression of γδT cells in the peripheral blood were not associated with age, gender (P ＞0.05). Conclusion The results suggested that the dysfunction of γδT had important function on the development of lung cancer.

We constructed several recombinant Escherichia coli strains to transform phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS system) and compared the characteristics of growth and metabolism of the mutants. We knocked-out the key genes ptsI and ptsG in PTS system by using Red homologous recombination in E. coli and meanwhile we also knocked-in the glucose facilitator gene glf from Zymomonas mobilis in the E. coli chromosome. Recombinant E. coli strains were constructed and the effects of cell growth, glucose consumption and acetic acid accumulation were also evaluated in all recombinant strains. The deletion of gene ptsG and ptsI inactivated some PTS system functions and inhibited the growth ability of the cell. Expressing the gene glf can help recombinant E. coli strains re-absorb the glucose through Glf-Glk (glucose facilitator-glucokinase) pathway as it can use ATP to phosphorylate glucose and transport into cell. This pathway can improve the availability of glucose and also reduce the accumulation of acetic acid; it can also broaden the carbon flux in the metabolism pathway.
Biological Transport ; Escherichia coli ; enzymology ; genetics ; Gene Deletion ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Glucose ; metabolism ; Phosphoenolpyruvate Sugar Phosphotransferase System ; genetics ; Zymomonas ; genetics

Objective To evaluate the changes and clinical significance ot the serum levels of homocysteine (Hcy) in gastric cardia cancer patients before and after surgery.Methods Serum Hcy concentrations of 102 patients with gastric cardia cancer (including 69 case of males) and 50 healthy human were measured by enzymatic cycling assay.Results Total Hcy levels were significantly increased in male patient group compared with the levels in control group (t =5.143,P =0.001).Hcy levels were significantly lower in postoperative group compared with preoperative group [(17.08±5.31) μmol/L vs (20.34±9.26) μmol/L,(14.07±4.87) μmol/L vs (20.34±9.26) μmol/L,P ＜ 0.05].Compared with Ⅳ stage group and other TNM stage groups,significantly lower levels of Hcy were detected in patients with gastric cardia cancer (t =2.306,3.285,P =0.030,0.002).Hcy levels in patients with gastric cardia cancer were also significantly higher than those in the tumor length ＜ 3 cm,3-5 cm and ＞ 5 cm groups (t =2.461,2.147,P =0.017,0.038).Multiple logistic regression analysis indicated a statistically significant association between serum Hcy concentration and gastric cardia cancer incidence (OR =1.136,95 ％ CI 1.010-1.278,P =0.033).Increasing serum Hcy levels were significantly associated with a decreasing risk of metastatic lymph node (OR =0.865,P =0.010).Conclusion Serum Hcy levels are directly associated with risk of male patients with gastric cardia cancer,and play important roles in the development of gastric cardia cancer.

Objective To investigate the role of HBXIP overexpression in the prognostic evaluation of breast cancer. Methods HBXIP expression was detected in the tissues of 60 breast cancer cases and 15 cases of ductal cancer in situ (DCIS) as well as in the adjacent non-tumorous tissues of 27 cases of breast cancer using EnVision immunohistochemical staining method. The correlations between the HBXIP overexpressions and the clinical pathological characters of the patients with breast cancer were also analyzed. Results The HBXIP proteins showed a major cytoplasmic staining pattern in breast cancer. The strongly positive rate of HBXIP was75.0%(45/60) in breast cancer, significantly higher than in DCIS (20.0%, 3/15) and adjacent non-tumorous tissues (14.8%, 4/27). High-level expression of HBXIP was correlated with late clinical stage, lymph node metastasis and HER-2 positive expression in breast cancer. Conclusions The expression level of HBXIP is closely related to the progression and prognosis of breast cancer. It might be a potential biomarker and therapeutic target of breast cancer.

Objective To investigate midkine (MK) level in serum of patients with esophageal squamous cell carcinoma (ESCC) before and after operation,detect the expression of MK protein in tissues and analyze its relationship with the malignant biological behavior of ESCC.Methods The MK levels in serum were detected by enzyme-linked immunosorbent assay in 30 healthy cases and 55 patients with ESCC when the day before operation and the tenth-day after operation.Then the expression of MK protein in 55 patients surgically removed ESCC were detected by immunohistochemistry using monoclonal antibodies against human MK.Results The average optical density of MK in serum of ESCC patients before operation was 0.1006±0.0624,0.0455±0.0155 in normal control group,0.0752±0.0267 in postoperative control group (t =6.203,P ＜ 0.001).The MK levels in serum of ESCC patients before operation were significantly higher than those after operation (t =4.357,P ＜ 0.001) and those in healthy controls (P ＜ 0.05).The expression of MK protein was no statistically significant correlation with tumor size (x2 =0.131),TNM stage (x2 =0.315) and lymph node metastasis (x2 =0.282) in ESCC (all P ＞ 0.05).But significant correlation was noted between the expression of MK protein and the vasculature involving of the cutting edge (x2 =4.223),degree of cellular differentiation (x2 =10.326),invasive extent of the carcinoma (x2 =20.556) (all P ＜ 0.05).Conclusion MK is overexpressed in ESCC patient' s serum and tissue.The high level of preoperative serum MK is caused by the tumor.

Objective To evaluate the changes and clinical significance of the serum level of homocysteine (Hcy) in esophageal cancer patients before and after surgery.Methods The serum Hcy concentrations of 168 patients with esophageal cancer and 50 healthy individuals were measured by enzymatic cycling.Results Total Hcy levels were significantly raised in patients group when compared with the levels in control group [(19.6±6.1) μmol/L vs (13.0±2.3) μmol/L,P =0.001].Hcy levels were significantly lower in postoperativepatients groups compared with preoperativegroup [(17.0±8.5),(15.8±6.4),(12.8±5.6) μmol/L vs (20.6±9.1) μmol/L,all P ＜ 0.05].Also,Hcy levels in patients with esophageal cancer were significantly higher than in the tumor length＞ 5cm group and the ＜3 cm,3-5 cm group [(23.6±9.6) μmol/L vs (18.1±6.3),(19.6±6.6) μmol/L,P =0.036,P =0.021].Compared with other T stage groups,significantly higher level of Hcy was found in T4 stage group [(29.5±1.7) μmol/L vs (18.5±6.9),(18.8±8.0),(20.6±8.8) μmol/L,all P ＜ 0.001].There was no significant difference between the mean Hcy concentrations of the node-positive and nodenegative group [(20.2±9.3) μmol/L vs (20.3±7.6) μmol/L,P =0.897].Compared with negative lymphaugial tumor-cells thrombus group,positive group had low Hcy level [(16.7±3.4) μmol/L vs (21.1±8.9) μmol/L,P =0.007].Conclusion These results showed that serum Hcy levels play important roles in the development of esophageal cancer.