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Objective: To analyze the prognostic factors of colorectal cancer patients with synchronous liver metastasis treated by simultaneous colorectal and liver resection., Methods: The clinical and follow-up data of 44 colorectal cancer patients with synchronous liver metastases who underwent simultaneous colorectal and liver resection from Jan. 1993 to Jan. 2003 were analyzed retrospectively. Survival rate was estimated by Kaplan-Meier method, and was compared using log-rank test. Prognostic factors were analyzed by multivariate Cox proportional hazards model., Results: The overall 1-, 3- and 5-year survival rates were 86.3%, 40.9% and 25.0%, respectively. The lymph node metastasis and vascular invasion by cancer cells from the primary tumour were found to affect prognosis significantly, while gender, age, tumor location, histopathological types, the number and distribution of liver metastases were not. Multivariate analysis revealed that the lymph node metastasis was the only independent prognostic factor., Conclusion: Simultaneous liver and colorectal resection can be performed and may achieve good outcome in colorectal cancer patients with synchronous liver metastases, especially in those without lymph node metastasis.

Objective: To investigate the experience on diagnosis and treatment of multiple adrenal aldosterone-producing adenomas (APA)., Methods: Eighteen cases of multiple adrenal APA were analyzed retrospectively, which were admitted from October 1992 to April 2006., Results: Adrenalectomy was performed for 4 cases of unilateral synchronous multiple APA, which were discovered with three adenomas by 3D-CT; bilateral tumor resection was performed for 6 cases of bilateral synchronous multiple APA. There were 8 cases of bilateral metachronous multiple APA, including 2 cases of ipsilateral recurrent adrenal APA after adrenal tumor removal, which underwent tumor resection. Another 6 cases were contralateral APA following adrenalectomy due to adrenal APA, and underwent tumor resection. After operation, the adrenal function seemed to be normal, and no recurrence had been found on follow-up., Conclusions: Unilateral multiple synchronous APA require adrenalectomy. Tumor resection should be performed for bilateral or asynchronous APA, and it is very important to preserve healthy adrenal tissue as much as possible. 3D-CT has much value on diagnosis of small APA, unilateral multiple synchronous APA and ipsilateral recurrent adrenal APA.

Objective: To investigate the surgical treatment and prognosis for recurrent colon cancer after curative resection., Methods: The clinical data of 102 recurrent colon cancer cases from January 1997 to December 2005 were analyzed retrospectively. Obtained data were analyzed by Statistical Package for the Social Sciences (Release 11.5, SPSS, Inc). The related factors were underwent chi2 analysis,survival analysis were estimated using the Kaplan-Meier method and compared using the Log-rank test. COX regression was used in multivariate analysis., Results: Univariate analysis revealed that obstruction of primary tumors, CEA level before reoperation, number of recurrence, time of recurrence, and reoperation type were significant statistically. COX regression analysis revealed that number of recurrence, reoperation type was the most important prognostic factor., Conclusion: The recurrent colon cancer still need active surgical treatment in order to prolong the survival time.

Objective: To summarize the surgical treatment experiece and to investigate the prognosis of the patients with ovarian metastasis from colorectal cancer., Methods: The data of 67 patients with synchronous or asynchronous ovarian metastasis from colorectal cancer surgically treated between January 1989 and December 2005 were collected and analyzed retrospectively using Statistical Package for the Social Sciences (Release 11.5, SPSS, Inc). Prognostic factors were analyzed using chi2 test. Survival analysis was estimated by the Kaplan-Meier method and compared using the log-rank test. Multivariate analysis was carried out by Cox regression., Results: The overall 1-, 3- and 5-year survival of these 67 patients was 71.0%, 18.7% and 9.2%, respectively. Univariate analysis revealed that the metastasis was confined in the ovary or pelvis only, unilateral/double ovarian metastasis, and operation mode were all statistically significant prognostic factors (P <0.05). Cox regression analysis showed that the operation mode was the most important prognostic factor (OR = 3.531, P <0.001)., Conclusion: Surgical treatment is still the most effective mode in the treatment for the ovary metastasis from colorectal cancer.

Adenovirus vectors are one of the most promising gene transfer systems. They are of great value for gene therapy because these vectors achieve temporal high-level transgene expression and high gene transfer efficiency. To meet increasing needs of adenovirus vectors for gene therapy programs, parallel development of efficient, scalable and reproducible production processes is required. Perfusion cultivation of 293 cells is one of the most commonly used methods to produce adenovirus vectors and it is suitable for industrialized production specially. Experimental studies had been carried out to produce recombinant adenovirus containing the green fluorescent protein gene (Ad-GFP) by perfusion cultivation of HEK-293 N3S cells in a 5L stirring bioreactors. Perfusion rate was 1-2 volume/day. To infect the 293 N3S cells with Ad-GFP at the density of (2-4) x 10(6) cells/ ml. The time of collecting cells was 48 hours post infection. After three rounds of freeze/thaw and centrifugation, the crude viral lysates were stored at--80 degrees C until use. Then to get the Ad-GFP products by 2 x CsCl-gradient purification. The purity of the products was determined by the A260/A280 ratio and a high performance liquid chromatography (HPLC) assay. The infective titer was determined by a TCID50 assay. The culture term was 10-12 days. The infectious titer, the number of virus particle and the ratio of infectious titer to virus particle for the product were 1.0 x 10(11) IU/mL, 1.68 x 10(12) VP/mL and 6.0% IU/VP respectively. The A260/A280 ratio was 1.33, and the purity determined by HPLC was 99.2%. The cell specific productivity was around 1000 IU/cell. By perfusion cultivation of 293 N3S cells in a 5L stirring bioreactors, we established the production process for Ad-GFP, which paves a way to produce other recombinant adenovirus for gene therapy.

To study the immunogenicity of Bordetella avium OMP, ultrasonic dispersion, TritonX-100 technique were used to extract Bordetella avium OMP. Its content was determined by Bradford method and it was detected by SDS-PAGE. Then OMP immunizing antigen was prepared and 1-day chicken was vaccinated by hypodermic inoculation, with the immunizing does of 0.3mL (OMP90microg), 0.5mL (OMP150microg), 0.8mL (OMP240microg), respectively. Results showed that content of Bordetella avium OMP is 300 g/mL, the best immunizing does is 0.5mL each one and Chickens can be protected against Bordetella avium at fetal dose if antibody titer is over 1:2(8). We can see from the antibody level detected by indirect ELISA that antibody can last long enough to help chickens pass susceptible period, so OMP has good immunogenicity. Results of this study lay good foundation for the development of monoclonal antibody to OMP, rapid diagnosis kit and subunit vaccine.

Conventional Bayesian analysis for proportions is not appropriate for proportions with a background effect, for example, for the proportion of correct responses in a sensory difference test using a forced-choice method with a guessing probability. The main difficulty is that it is not reasonable to assume that a proportion with a background effect has a beta prior distribution. A generalized posterior distribution for proportions is derived in this paper. It includes the standard beta posterior distribution for a proportion without background effect as a special case. Bayesian inference and Bayesian sample size determination for proportions based on the generalized posterior distribution are discussed.

Objective: To investigate the surgical treatment of ovarian metastasis from colorectal cancer., Methods: The clinical data of 62 cases suffering from ovarian metastasis from colorectal cancer, collected from Jan. 1990 to Dec. 2005, were analyzed retrospectively., Results: The median survival time of 62 colorectal cancer patients with ovarian metastasis was 23 months. The median survival time of 19 patients with simple ovary metastasis was 31 months, while that of 43 patients with ovary and other organ metastasis was 21 months. The median survival time of 28 patients (45.2%) treated with radical resection was 31 months, while that of 34 patients (52.8%) treated with palliative resection was 20 months, the difference between two groups was significant. Fifty-one patients (82.3%) were treated with double-sided ovarian resection, and 42 of them (17.7%) received hysterectomies at the same time. Eleven patients received one-sided ovarian resection, and 8 of them were resected the metastatic ovaries on the other side in 3 to 10 months., Conclusions: The patients with ovarian metastases from colorectal cancer need double-sided ovarian resection, and radical resection is able to prolong the survival time.

Objective: To evaluate the efficacy and safety of postoperative adjuvant chemotherapy with imatinib in gastrointestinal stromal tumor(GIST) patients who had high risk of recurrence., Methods: A prospective, open-label, multi-center trial conducted in sixteen teaching hospitals in China was carried out. The criteria of the enrolled patients included age more than 18 years old, CD117 positive GIST, tumor size more than 5 cm, pathological mitosis counts more than 5/50 HPF, and treatment beginning within 4 weeks after complete resection and with imatinib (400 mg, once a day) for at least 12 months. The 1, 3 year recurrence rates, disease free survival, overall survival rate and quality of life were evaluated., Results: From Aug. 16th 2004 to Sep. 13th 2005, there were totally 74 patients screened and 57 patients (34 men, 23 women) enrolled in the imatinib treatment group. The primary tumors were located in the stomach in 50.9%, the small intestine in 38.6% and the colorectum in 10.5% of the cases. All the patients received radical resection. Until the cut-off date of interim analysis, there was no evidence of tumor relapse or metastasis in all patients and no death was reported either. Among the 57 enrolled patients with intention to treat(ITT), twelve patients finished the protocol (per protocol, PP). The disease free survival was (268.3 +/-120.2) d in ITT analysis, and (396.7+/-38.2) d in the PP analysis. The incidence of adverse effect was 44.4% . The score in quality of life showed no statistically significant difference between the baseline visit and the follow-up visits., Conclusion: Imatinib is a promising postoperative adjuvant chemotherapy in GISTs patients with high risk of recurrence, and the adverse effects are receivable.

Objective: To investigate the surgical treatment for recurrent colon cancer after radical resection., Methods: Clinical data of 87 cases with recurrence colon cancer after radical resection from Jan. 1999 to Dec. 2005 were analyzed retrospectively., Results: The resection rate of recurrent colon cancer was 74.7% . 55.2% (48/87) of the cases received radical resection,and the median survival was 49 months,while 19.5% (17/87) received palliative resection with a median survival of 24 months, 25.3% (22/87) only exploration or by- pass operation with a median survival of 10 months. There were significant differences in survival among the different surgical treatments (P=0.003)., Conclusion: The resection rate of recurrent colon cancer is high,and reoperation can prolong the survival of such patients.

Objective: To investigate the characteristics of lymph node metastasis and prognosis of T1/T2 rectal carcinoma., Methods: The clinical data of 241 patients with T1 or T2 rectal carcinoma were retrospectively analyzed. The factors relative to lymph node metastasis were analyzed using Chi-square test. The survival data were analyzed using Kaplan-Meier method. The factors influencing survival were analyzed using univariate (Long-rank) and multivariate (Cox model) methods., Results: Of the 241 patients, 132 received Mile's operation and 109 underwent sphincter preserving operation. The over-all lymph node metastasis rate was 22.0% (53/241). The lymph node metastasis was significantly correlated with histological differentiation as revealed by Chi-square test. The over-all 5-year survival rate for the whole series group was 91.5%. Univariate analysis revealed that tumor histological type, intramural infiltration, differentiation, lymph node metastasis, radiation therapy were significant predictors of survival; however, only intramural infiltration was the most important prognostic predictor by multivariate analysis., Conclusion: Even though lymph node metastasis can be observed either in T1 or T2 rectal carcinoma, histological differentiation is significantly related to the lymph node metastasis. As radical resection achieve better survival than local resection, it should be suggested as the chief treatment for T1/T2 rectal carcinoma.

Objective: To investigate the clinicopathologic factors related with recurrence and prognosis after surgical resection for I stage lower rectal carcinoma., Methods: The related clinicopathologic factors for recurrence and prognosis of 166 patients with I stage lower rectal carcinoma after surgical resection were retrospectively analyzed using univariate and multivariate methods., Results: A total of 138 patients with I stage lower rectal carcinoma received radical resection according to the operative rules of total mesorectal excision (TME). Ninety-three patients received abdominoperineal resection (APR) operation, 45 patients received sphincter preserving operation, and 28 patients received local excision. The local recurrence rates were 6.5% (6/93), 2.2% (1/45), 17.9% (5/28), respectively . Histological differentiation and operative procedures were associated with local recurrence. The 5-year survival rates were 91.1% in APR group, 95.5% in sphincter preservation group and 82.6% in local resection group. Univariate analysis revealed that histological differentiation and local recurrence were correlated with prognosis. Multivariate analysis revealed that local recurrence was the most important prognostic factor for I stage lower rectal carcinoma., Conclusions: Radical resection of I stage lower rectal carcinoma has low recurrence rate and better prognosis. Sphincter preserving operation and local excision must be strictly selected in proper patients.

Objective: To explore the mechanism of reversal of multidrug resistance in renal carcinoma cells by protein kinase C inhibitor., Methods: RT-PCR, Western blot and inverted fluorescent microscopy were used to determine the expression of PKCalpha and MDR related gene MDR1, MRP1, LRP in RCC cells transferred by PKCalpha cDNA. Also effects of activator and inhibitor of PKC in combination with adriamycin on multidrug resistance in RCC cells were evaluated by MTT., Results: The results of semi-quantitative RT-PCR analysis showed that the expression level of MDR1 was higher in RCC cells transferred by PKCalpha cDNA than in RCC cells, the reversal effectiveness of PKC inhibitors in combination with adriamycin (ADM) was apparently favorable. IC(50) of ADM in 786 - 0 cells was 7.8015e(-7) (5.7046e(-7) to 1.0669e(-6)); IC(50) of ADM in PKCalpha/786 - 0 cells was 1.6588e(-6) (1.1621e(-6) to 2.3677e(-6)); IC(50) of ADM in combination with PMA in PKCalpha/786 - 0 cells was 2.6794e(-6) (2.0521e(-6) to 3.4983e(-6)); IC(50) of ADM in combination with calphostin C in PKCalpha/786 - 0 cells was 9.2506e(-8) (5.9337e(-8) to 1.4422e(-7))., Conclusion: PKC inhibitors can reverse multidrug resistance in renal carcinoma cells in vitro via changes of expression of MDR1.

Produced by and purified from Taxus brevifolia, Taxol (paclitaxel) has become a widely used cancer drug in clinic. Due to the rapid growing market, current industrial production of taxol by semi-synthesis that consumes large amount of Taxus trees cannot meet the requirement of the market. The discovery of taxol-producing fungus Taxomyces andeanae, an endophyte of T. brevifolia, by Stierle et al (1993), paves a new way to the production of the drug, i.e. employing large-scale fungal fermentation to make Taxol at lower cost and yet higher yield. This review discusses the present problems in taxol production in pharmaceutical industry, the finding and research progress on taxol-producing fungi, and the potential application of fungal fermentation to manufacture this important drug.

Objective: The combination of oxaliplatin (L-OHP), 5-fluorouracil (5-Fu) and folinic acid (FA), being one of the effective regimens for advanced gastric cancer, is used in form of chronomodulated chemotherapy for advanced gastric cancer to investigate its efficacy and safety., Methods: Twenty-six patients received a 4-day chronomodulated infusion of L-OHP, 5-Fu and FA. L-OHP (25 mg.m(-2).d(-1)) infused from 10:00 am to 22:00 pm, and followed by 5-Fu (600 mg.m(-2).d(-1)) and FA (300 mg.m(-2).d(-1)) from 22:00 pm to 10:00 am for 4 days using a multichannel programmable pump, every 2 weeks as an cycle for at least 2 cycles., Results: Twenty-six patients with previously untreated advanced gastric cancer were eligible. Two complete and 13 partial remissions were observed with an overall response rate of 57.7%. Stable disease was observed in 6 patients (23.1%) and progressive disease in five (19.2%). Four of these patients underwent surgery. The median remission time was 3.5 months and time to tumor progression (TTP) was 4.5 months. The median overall survival time was 8 months. A total of 80 cycles were given without any grade 4 toxicity observed, but grade 3 thrombocytopenia (1.3%) and mucositis (1.3%) in one patient, two grade 3 neutropenia (2.5%) and nausea/vomiting (2.5%) in 2., Conclusion: Chronomodulated intravenous chemotherapy of oxaliplatin, 5-fluorouracil and folinic acid is effective and safe in the treatment of advanced gastric cancer.

Severe depletion of myocardial energy and antioxidant resources during cardiac arrest culminates in electromechanical dysfunction following recovery of spontaneous circulation (ROSC). A metabolic fuel and natural antioxidant, pyruvate augments myocardial energy and antioxidant redox states in parallel with its enhancement of contractile performance of stunned and oxidant-challenged hearts. This study tested whether pyruvate improves post-arrest cardiac function and metabolism. Beagles were subjected to 5 min cardiac arrest and 5 min open-chest cardiac compression (OCCC: 80 compressions min(-1); aortic pressure 60-70 mmHg), then epicardial dc countershocks (5-10 J) were applied to restore sinus rhythm. Pyruvate was infused i.v. throughout OCCC and the first 25 min ROSC to a steady-state arterial concentration of 3.6+/-0.2 mM. Control experiments received NaCl infusions. Phosphocreatine phosphorylation potential (approximately PCr) and glutathione/glutathione disulfide ratio (GSH/GSSG), measured in snap-frozen left ventricle, indexed energy and antioxidant redox states, respectively. In control experiments, left ventricular pressure development, dP/dt and carotid flow initially recovered upon defibrillation, but then fell 40-50% by 3 h ROSC. ST segment displacement in lead II ECG persisted throughout ROSC. Approximately PCr collapsed and GSH/GSSG fell 61% during arrest. Both variables recovered partially during OCCC and completely during ROSC. Pyruvate temporarily increased approximately PCr and GSH/GSSG during OCCC and the first 25 min ROSC and enhanced pressure development, dP/dt and carotid flow at 15-25 min ROSC. Contractile function stabilized and ECG normalized at 2-3 h ROSC, despite post-infusion pyruvate clearance and waning of its metabolic benefits. In conclusion, intravenous pyruvate therapy increases energy reserves and antioxidant defenses of resuscitated myocardium. These temporary metabolic improvements support post-arrest recovery of cardiac electromechanical performance.

Objective: In order to get a better understanding of the mechanism of DNA initiating specific cytotoxicity T lymphocyte (CTL) immune response, we observed the local infiltration in injection site at different time after DNA immunization and further studied the characters of infiltration cells., Methods: The local infiltration in injection sites was observed by HE staining at 6 h, 12 h and 1-6 days after DNA immunization. Infiltration cells were further studied with the mice dendritic cells marker CD205, T lymphocyte marker CD4 and CD8 by immunohistological methods., Results: phAFP injection into mouse muscle led to a local infiltration. Infiltrates reached peak at 3-6 days after injection and appeared in several discrete clusters within the muscles. The predominant cell types found in infiltrates were macrophage-like cells and lymphocytes. Immunohistological staining revealed CD205+ and CD4+ cells. CD4+ cells could be detected in any of the injected muscles. CD205+ cells were only seen at 24 h to day 3, appearing later and disappearing earlier than CD4+ cells. However, CD8+ immunostaining positivity could not be found., Conclusion: Our findings indicate that the character of local infiltration after DNA immunization is different from the inflammation of injection. The recruitment of CD205+ to the injection site suggests that DCs and Th cells play an important role in DNA immunization.

A commonly used method of estimating population sensitivity is the so-called averaged d' method. In this method, the arithmetic mean of a set of individual d' is usually taken as a population sensitivity estimator. This practice ignores the fact that the individual d' itself is an estimator with an inherent variance. For observations with different levels of precision, the arithmetic mean is not the best estimator of a population parameter. It may lead to an estimate with a large variation. Another fact, which is often ignored, is that the variance of individual d' involves both between- and within-subject variations in a random effects model when population sensitivity and its level of precision are estimated. Failing to account for both components of variance leads to an underestimate of variation and an overestimate of precision for the estimator. In this paper a lognormal distribution rather than a normal distribution is assumed for individual sensitivity. An iterative weighting procedure is proposed for estimating population sensitivity on the log scale on the basis of a random effects model. An ordinary weighting procedure is proposed for estimating group sensitivity on the log scale on the basis of a fixed effects model. The levels of precision of population and group sensitivity estimators are also given. Numerical examples illustrate the estimation procedures.

Background & Objective: Abdominal recurrence from exfoliated cancer cells contributes a lot to treatment failure of advanced gastric cancer. Intraperitoneal chemotherapy, which has been proved effective in eliminating exfoliated cancer cells in abdominal cavity, is a hot topic on treatment of gastric cancer. This study was to explore application of combined therapy of intraoperative hypotonic peritoneal chemo-hyperthermia and early postoperative intraperitoneal chemotherapy to gastric cancer., Methods: A total of 156 gastric cancer patients were randomized into 3 groups, and underwent the combined therapy (treatment group 1), intraoperative chemotherapy (treatment group 2), and peritoneal lavage with distilled water (control group), respectively., Results: The 2-year survival rate of treatment group 1 was significantly higher than that of control group (88.4% vs. 65.2%, P < 0.05). The 3-year survival rate of treatment group 1 was significantly higher than those of treatment group 2, and control group (71.1% vs. 50.0%, and 45.6%, P < 0.05). Occurrence of liver metastasis was significantly lower in treatment groups 1 and 2 than in control group (7.7%, and 10.2% vs. 27.3%, P < 0.05)., Conclusions: Combined therapy of intraoperative hypotonic chemo-hyperthermia and early postoperative intraperitoneal chemotherapy is effective for gastric cancer. Intraperitoneal chemotherapy can be used to prevent postoperative liver metastasis of gastric cancer.

Objective: To investigate the effect of replication-competent adenovirus-mediated interleukin-12 gene to chemotherapeutic sensitivity on gastric cancer cell., Methods: Replication-competent adenovirus and replication-competent adenovirus- mediated interleukin- 12 gene was constructed and expanded separately. The mortality of gastric cancer cell caused by the CNHK200- mIL- 12, Onyx- 015 in combination with different dosages of chemotherapeutic agents were evaluated by MTT assay at the same viral titer with a series of different dosages of chemotherapeutic agent,or at a series of different viral titers with the same dosage of chemotherapeutic agent. The curative effect to the xenografts gastric tumor in nude mouse was also observed by two viruses solely or together with 5-Fu., Results: The lytic activity of replication-competent adenovirus to gastric cancer cell line SGC-7901 was relatively poor at MOI value of 0.5, but it could be improved significantly when combined with chemotherapeutic agents of ADM, 5-Fu or CAP compared to the simple chemical therapy (P< 0.05). Chemotherapeutic agent 5- Fu could not effectively kill SGC-7901 when used at a relatively low dosage of 10microg/ml,whereas its activity could be improved when combined with a replication-competent adenovirus,and the killing rate was much higher than that with replication-competent adenovirus solely (P< 0.05). The gastric tumor xenografts was prevented and killed by replication adenovirus solely or combined with 5-Fu., Conclusion: The replication- competent adenovirus- mediated interleukin- 12 gene can increase the chemotherapeutic sensitivity on gastric cancer cell. There is synergetic effect between the replication adenovirus and the chemotherapeutic agents in killing gastric cancer cell.

A competitive enzyme-linked immunosorbent assay (ELISA) technique was evaluated for quantifying titers of imidacloprid in xylem fluid extracted from Vitis vinifera L. grapevines that were treated with systemic applications of the neonicotinoid insecticide Admire. Evidence of matrix effects, factors that compromise the precision and accuracy of the ELISA, was present in assays with undiluted xylem fluid. These effects could be eliminated by dilution of extracts in water, resulting in a lower sensitivity of the assay of 4 microg liter(-1). In a field trial conducted in a commercial vineyard, there was an excellent correlation between Admire application rates and xylem fluid concentrations of imidacloprid. At an Admire application rate of 1.17 liter ha(-1) (16 fl oz per acre), uptake of imidacloprid into vines was rapid. Imidacloprid was consistently detected in the xylem for up to 3 mo after application at concentrations known to be effective at managing populations of the sharpshooter Homalodisca coagulata Say, an important vector of Xylella fastidiosa Wells in California vineyards. The ELISA is a sensitive technique that can be used to study the behavior of systemic insecticides within crop systems and their impact on pest populations.

Objective: To construct anti-human AFP single chain fragment variable (ScFv) gene, transform it into BL-21 (DE3) E. coli for expression, and identify its bioactivity., Methods: VH and VL genes of anti-human AFP monoclonal antibody were cloned by RT-PCR from hybridoma. The ScFv gene was spliced by sequence overlap extending (SOE) PCR, and then it was ligated into pGEM-T vector to be identified by endonuclease digestion, PCR and sequencing. ScFv gene was cloned into pET32 (a+) vector and transformed into BL-21 (DE3) E. coli. The positive clones were screened out by IPTG induction, and the ScFv antibody was purified to be identified by SDS-PAGE and competitive inhibition ELISA test., Results: The VH DNA consisted of 339 bases, coming from the mouse IgG gamma chain. The VL DNA consisted of 312 bases, coming from the mouse IgG kappa chain. The VH and VL genes were spliced by 45 bases coding a (G4S)3 flexible linker. The ScFv gene consisted of 696 bases. The ScFv antibody expressed by BL-21 (DE3) fused with TrxA tag protein and formed inclusions. The relative molecular mass of TrxA-ScFv fusion protein is about 40 x 10(3) and that of ScFv is about 24 x 10(3). The ScFv antibody has excellent activity tested by competitive inhibition ELISA, the TrxA-ScFv could inhibit about 41% of the McAb to bind antigen and ScFv could inhibit about 53%., Conclusion: We have successfully constructed an anti-human AFP ScFv gene with 696 bases; it can express in BL-21 with high activity.

Objective: To clone the variable region genes of the monoclonal antibody (McAb) against human heterogeneous nuclear ribonucleoprotein A2/B1 (HnRNPA2/B1), ligate them to assemble single chain Fv (ScFv) gene and express in Escherichia coli., Methods: The specificity of the anti-HnRNPA2/B1 McAb 3E8 to synthetic HnRNPA2/B1 peptide, HnRNPA2/B1 protein in lung cancer cells were examined by dot-immunobinding assay, Western blot and immunohistochemistry. The variable region genes of heavy chain (VH) and light chain (VL) were amplified from hybridoma cell by reverse transcription-polymerase chain reaction(RT-PCR), and then were linked by a linker peptide using SOE-PCR (splicing by overlap extension-PCR) to construct recombination ScFv gene. The latter was cloned into the expression vector pET28 (a+) and expressed in E coli BL21. The expressed product was identified by SDS-PAGE and competitive ELISA inhibition test., Results: It was shown that the McAb combined specifically with synthetic HnRNPA2/B1 peptide and HnRNPA2/B1 protein in three lung cancer cells. The cloned VH gene and VL gene were 345 bp and 309 bp respectively and were linked successfully to obtain ScFv gene. The ScFv protein was expressed in the form of inclusion body, with molecular weight of 28,000 and immunoreactivity to HnRNPA2/B1., Conclusion: VH gene, VL gene and ScFv gene of anti-HnRNPA2/B1 antibody were cloned, constructed and functionally expressed in E coli. These results provide the experimental basis for elucidating the role of HnRNPA2/B1 in lung cancer.

Objective: To construct AFP-expressing plasmid and study the expression in eukaryotic cells., Methods: Total RNA was isolated from fetal liver tissue. Full-length human AFP cDNA was obtained by RT-PCR amplification and then recombined into eukaryotic expression plasmid pcDNA3.1. The AFP sequence of the recombinant plasmid phAFP was determined. The AFP expressions in CHO transfected with phAFP and in muscle after injection phAFP were investigated by immunohistological methods., Results: The restriction endonuclease mapping of recombinant plasmid showed 1.8 kb fragment as well as full-length human AFP cDNA, and the sequence is consistent with that from GenBank. The phAFP was successfully expressed in CHO and in muscle tissues., Conclusion: These results suggested that AFP can be used as a target gene of DNA vaccine in heptocellular carcinoma therapy.

Objective: To verify the feasibility and effect of biotherapy instituted via pelvic retroperitoneal space on gynecological cancer., Methods: Injecting IL-2 (and/or) 5-Fu through a tube installed in the pelvic retroperitoneal space. Counting the subpopulation of T cell and NK of lymph-nodes of pelvis after the drugs being by FCM., Results: The numbers of CD3+, CD4+, CD8+, CD25+ and NK cells in treatment group were significantly higher than those in the control group. And the numbers of these cells in the IL-2 + 5-Fu group were significantly higher than those in the 5-Fu group. The CD25+ and NK cell numbers in the IL-2 group were significantly higher than those in the 5-Fu group (P < 0.05)., Conclusion: The IL-2 injected via pelvic retroperitoneal space can promote the activity, development and infiltrating of T cell and NK cell in the tumor tissue.

Background & Objective: X-ray radiography is the first choice for detecting chest diseases. However, "visual obstacles" caused by the overlaps between ribs and lung in chest radiography affects the observation and analysis of chest diseases. The objective of this study was to investigate the value of direct digital radiography (DR) dual-energy technique in diagnosis of chest disease located in the overlaps between ribs and lung., Methods: Thirty-five cases of chest tubercular diseases located in the overlaps between ribs and lung were examined using direct digital radiography (DR) dual-energy technique., Results: Ratings of dual-energy technique radiography in detecting chest tubercular diseases were 94.3% whereas the ratings of DR radiography were 45.7%. Bone and soft tissue can be separated in the radiography using dual-energy technique., Conclusion: DR dual-energy technique not only increases the visibility towards chest tubercular disease, but also discriminates the location (ribs or lung) of chest tubercular disease.

Objective: To investigate whether basic fibroblast growth factor (bFGF) can induce the proliferation, invasion and angiogenesis of ovarian cancer or not., Methods: Human ovarian cancer cell lines SKOV(3) 1 x 10(4)/ml were plated in 24-well dishes, bFGF at 5, 10,15 and 20 ng/ml was added and crystal violet staining was given daily for 8 days, cell numbers were counted by determining OD490. SKOV(3) cells were plated in the center of 50% extra cellular matrix gel, bFGF at 5 and 10 ng/ml was added and the migration distance of cells was measured daily. SKOV(3) 5 x 10(7)/ml were transplanted to BALB/c nude mice subcutaneous. One week later, bFGF, bFGF-MAb or 0.9% nature sodium was injected subcutaneously surrounding the tumor twice a week. Eight weeks later, the experiment ended and the volume of the tumors were measured. Intratumoral microvessel density (MVD) was measured by immunohistochemistry staining for factor VIII., Results: bFGF at 0-10 ng/ml could stimulate the proliferation of SKOV(3) concentration dependently (P<0.05). On the fifth day, the cell proliferation in 10 ng/ml group was 121% above control. bFGF could stimulate the invasion of SKOV(3) concentration dependently (P<0.05). On the seventh day, the migration distance in 5 ng/ml group was 1.16 cm and 153% above control, and that in 10 ng/ml group was 1.86 cm and 245% above control. The average volume of transplanted tumors and MVD in bFGF group were 180% and 146% above control respectively those in bFGF-MAb group were 63.7% and 62.8% above control respectively., Conclusion: bFGF can stimulate proliferation, invasion and angiogenesis of ovarian cancer markedly; bFGF-MAb can inhibit the angiogenesis and growth of ovarian cancer.

Objective: To investigate the effect of plasmid pUDKH carrying human hepatocyte growth factor (HGF) gene on rat acute limb ischemia and to find the lowest effective dosage., Methods: Ligation of femoral artery of one hindlimb in Wistar rats was performed. The rats were randomly divided into 5 groups of 10 rats: pUDKH 50 microg, 100 microg, 200 microg, and 400 microg groups and a vacant vector pUDK group (control group). The plasmids were injected once directly into the ischemic limb muscle (5 sites around ligation position) immediately after ligation. At day 10, the muscles were removed and stained with H.E. to assess histologically the blood vessel formation. HGF expression was detected in the muscle tissue of another 12 rats on days 3, 5, 10, 14, 21, 30 after injection of pUDKH or pUDK, 200 microg each per animal., Results: HGF expression was detected clearly in muscle tissue on days 3, 5, 10, 14 in pUDKH groups. In HGF-transfected animals (except for 50 microg group) neoformation of blood vessels in muscles and soft tissues was found, while it was not seen in controls. By semi-quantitation of the degree of vessel formation, significant differences between pUDKH groups (0.96 +/- 0.07, 1.97 +/- 0.91, 2.26 +/- 1.00 for 100 microg, 200 microg, 400 microg, respectively) and the control group (0.27 +/- 0.04) (P < 0.05) were noted. In addition, injection of pUDKH could alleviate the severity of degeneration of muscular tissue due to ischemia., Conclusion: Human HGF gene therapy is effective in treating rat acute limb ischemia with the lowest effective dose of 100 microg.

Aim: To assess the validity of local excision for the early stage low rectal cancer as an effective treatment alternative to radical resection., Methods: A retrospective medical chart review was done in 47 patients with early stage low rectal carcinoma who underwent local excision from November 1980 through November 1999 at Cancer Hospital of Chinese Academy of Medical Sciences (CAMS). The patients were treated by either transanal (40 cases), trans-sacral (5 cases), or trans-vaginal (2 cases) excision of tumors and no death was related to surgery. Sixteen patients received postoperative radiotherapy., Results: T1 and T2 lesion was found in 36 (76.6 %) and 11 patients (23.4 %) respectively. The overall local tumor recurrence rate was 14.9 % (7/47), with an average recurrence time of 21 months. Among these 7 recurrent patients, there were 4 T1 and 3 T2 lesions. Microscopically, the surgical incisal margin was negative in 45 (95.7 %) and positive in 2 patients (4.3 %); Both of the later had developed local recurrence. The overall 5-year survival rate was 91.7 %, in which there were 94.4 % for T1 and 83.3 % for T2 tumors. T stage, intravessel tumor thrombosis, lymphocytic infiltration and histological grade were not found to be significant by related to the local recurrence and survival (P>0.05)., Conclusion: Local tumor excision was a safe procedure for the treatment of early stage low rectal carcinoma with minimal morbidity and mortality, which might serves as one of the primary surgical treatment methods for the disease of this kind.

Objective: To study the clinical features, treatment and diagnosis of parapelvic cyst., Methods: Twenty-three patients of parapelvic cyst of the kidney were reviewed retrospectively. Fourteen cases (61%) complained of lumbar pain or discomfort, and 4 patients (17%) accompany hematuria and hypertension., Results: In 15 patients receiving surgery, 2 were treated by nephrectomy, one by radical nephrectomy for misdiagnosis. Postoperative diagnosis confirmed a cyst. Eight patients were treated conservatively for cyst being small and without clinical symptoms. Nineteen cases were followed up for 0.5 - 12.0 years., Conclusions: Ultrasonography and CT scan are the main diagnostic methods. Enhanced CT is extremely helpful in differential diagnosis of hydronephrosis. Surgical management is suitable for big cysts, lumbar pain, hematuria, hypertension and other complications.

Variance estimates of d' are derived toward the same-different method based on Taylor-series expansion with one and two variables. The variance estimates can be used for statistical comparison of d's obtained from various discrimination paradigms. Formulas and tables for estimating variance of d' for the method are provided. One S-PLUS program, which can produce both d' and variance of d', is also provided.

Objective: To explore the effects of Zhikuofang, a TCM prescription, and Ofloxacin on the inflammation and cytostatics of the airway model of bronchiectasis., Method: The airway model of bronchiectasis (AMB) was set up and infused with Ps. Aeruginosa. A comparison between the effects of Zhikuofang and Of loxacin on the AMB was made., Result: Zhikuofang is better than Ofloxacin in following aspects: lowering the density of inflammation cells in blood, decreasing the volume of tracheal secretion and inhibiting the cytostatics (IL-8 and TNF-alpha) of the trachea tissue, but Ofloxacin is more effective in diminishing the amount of bacteria in trachea flushing liquor. There was no marked difference between them in their histopathy effects on the trachea., Conclusion: Zhikuofang probably plays antiphlogistic and bacteriostatic effects by inhibiting the IL-8 and TNF-alpha, resisting secretion, decreasing the inflammation cells and resisting inflammation of trachea.