“小产权房”问题解决的法律分析

'小产权房'问题成为近年来中国房地产业的一个不能触摸的肿块,正如医生对一个不能确诊的瘤子不能决定到底是良性还是恶性,担心打开以后难以控制局势,但不想办法解决这个瘤子,身体的危险又是非常明显的。'小产权房'问题亦如斯,彻底地解决将涉及社会不稳定;如果存而不议,已经蹒跚步入市场的中国房地产业将变得更为畸形,更多的利益体将利用法律法规的灰色地带继续牟取更多的暴利。“小产权房”要解决,又怎样解决呢?

Dynamic Transportation of Endophytic Rhizobia in Plants during Growth Development of Alfalfa

[ Objective] The aim of this study was to investigate the dynamic distribution of endophytic rhizobia in different plant parts during seed development of alfalfa and the carrying mechanism of endophytic rhizobia from alfalfa seeds. [ Method] Root, stem, leaf, flower, pod and seed from two varieties of alfalfa in different development stages were used for surface disinfection, and then the present site and quantity change of rhizobia in alfalfa were studied by the dilution - plate method. [ Result] The number of rhizobia from underground plants in the same stage was as follows： main root 〈 lateral root 〈 hair root. The number of rhizobia from main root and hair root in different stages was as follows： bearing pod stage 〉 branching stage 〉 florescence, while that from lateral root was as follows： branching stage 〉 florescence 〉 bearing pod stage. The distribution of endophytic rhizobia from aerial plants was as follows： few rhizobia was found in stem during branching stage and bearing pod stage, which accounted for 155 cfu/g and 125 cfu/g respectively. Rhizobia mainly distributed in flower bud during branching stage accounting for 9 400 cfu/g, while only in ovary wall during squaring stage accounting for 18 cfu/5 flowers, and few in pollen （26 cfu/5 flowers）, fertilized ovules （26 cfu/50 grains）, receptacle （15 cfu/5 flowers） and ovary wall （95 cfu/5 flowers） during florescence. Rhizobia existed in pod peel （3 550 cfu/5 pods） and immature seed （1 135 cfu/50 grains） during bearing pod stage, and greatly distributed in seed coat of mature seeds （1 435 cfu/50 grains）, but only few in embryo （32 cfu/50 grains） and cotyledon （22 cfu/50 grains）. [ Cenclusion] There is a significant difference in the distribution of endophytic rhizobia from different parts of aerial alfalfa, which only exists in the parts directly related to seed development.

腮腺炎合并脑膜脑炎急性期脑脊液中IL-1、IL-6、IL-8、TNF-α的变化及其意义

为探讨细胞因子在病毒性脑炎发病中的作用及意义,对30例腮腺炎合并脑膜脑炎患儿脑脊液测定了IL-1、IL-6、IL-8(白细胞介素-1、6、8),TNF-α(肿瘤坏死因子),结果发现急性期腮脑患儿脑脊液中上述细胞因子均较对照组明显增高(P<0.01),说明腮腺炎合并脑膜脑炎的发病与IL-1、IL-6、IL-8、TNF-α密切相关。

Recombination Mutant Human Tumor Necrosis Factor Combined with Chemotherapy in the Treatment of Advanced Cancer

Objective: Past studies showed that tumor necrosis factor (TNF) assisted anti-tumor treatment and intensified the sensitivity of chemotherapy. However its clinical application has been curbed because of its low purity, high dosage, and strong toxicity. The objective of present study is to evaluate the therapeutic effects and adverse reactions of recombinant mutant human tumor necrosis factor (rmhTNF) combined with chemotherapy in patients with advanced malignant tumor. Methods: 105 patients with advanced malignant tumor were randomly divided into trial group, 69 patients, and control group, 36 patients. rmhTNF was injected intramuscularly to the trial group at a dose of 4×106 U/m2, from the 1st to 7th days, the 11th to 17th days combined with chemotherapy course. The chemotherapy plan was as follows: CAP for patients with the NSCLC; FAM for patients with gastric cancer; FC for patients with colorectal cancer. One treatment cycle lasted for 21 days and two cycles were scheduled. The control group was given only the same chemotherapy as the trial group. Results: In the trial group there was 1 CR case and 12 PR cases, and the response rate was 13/69 (18.84%); in the control group 1 PR case, the response rate 1/36 (2.78%). The response rate in the trial group was significantly higher than that in the control group (P=0.022). The response rate for NSCLC in the trial group was 8/17 (47.06%), and 1/6 (16.67%) in the control group. The response rates for gastric cancer and colorectal cancer in the trial groups also were higher than those in the control groups. After the treatment the KPS was 89.00±9.92 in the trial group, and 84.17±8.84 in the control group, with a significant difference between the two groups (P=0.028). The adverse reactions of rmhTNF injection included: pain in the injection area, chill, hardening and swelling and redness in the injection area, fever, ostealgia and myosalgia, and cold-like symptoms. All these adverse reactions were mild and bearable. Conclusion: The administration of rmhTNF in combination with general chemotherapy is an effective and secure means in treating advanced malignant tumor.

Changes of Src-suppressed C kinase substrate expression in cytokine induced reactive C6 glioma cells

Objective To investigate effect of tumor necrosis factor-or （TNF-α） on the Src-suppressed C kinase substrate （SSeCKS） in C6 glioma cells. Methods Cultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-α （2 ng/mL） for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-α （0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL） for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS＇s subcellular localization. Results TNF-α induced rapid phosphorylations of protein kinase C （PKC） substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-α induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220. Conclusion TNF-α activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.

Expression Profiles of TRAIL Receptors and Their Clinical Significance in Human Hepatocellular Carcinoma

Objective To investigate the expression profiles and their clinical significance of TRAIL receptors (TRAILR) in human hepatocellular carcinoma (HCC). Methods The expression profiles of TRAILR were determined in 60 samples from hepatocellular carcinoma, 20 from normal liver tissue and two HCC cell lines HepG2, SMMC-7721 by in situ hybridization. Results Both DR4 and DR5 were present in all HCC tissues as well as normal hepatic tissues. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples (higher DR expression level and lower DcR expression level) were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. The expression level of DR was correlated with HCC differentiation and stage. The weaker expression was more commonly found in HCC with poor differentiation and late stage, while the stronger expression was more common in HCC with middle to high-differentiation and early stage. No relationship was found between DR and gender, age, negative or positive HBsAg, tumor size, grade or metastasis. Multidrug resistance cell lines expressed lower level DR. Conclusion TRAILR expression was prevalent and discrepancy of receptor types was exited in HCC. Loss of DcR1 may contribute for TRAIL therapy for HCC. Key words TRAILR - apoptosis - hepatocellular carcinoma Supported by the Major Fundation of Ministry of Health, NO. 2001–2003

Akt1/p27^(kip1) Pathway Mediates Inhibition of LXA_4 on TNF-α-induced Proliferation of Rat Mesangial Cells

Objective:To examine whether lipoxin A 4(LXA 4) has an inhibitory effect on tumor necrosis factor-α(TNF-α)-induced proliferation of glomerular mesangial cells of rat, and explore the molecular mechanisms of signal pathway in LXA 4 actions. Methods: Glomerular mesangial cells of rat were cultured and treated with TNF-α(10 ng/ml), with or without preincubation with LXA 4 at different concentrations. Cell proliferation was evaluated by monotetrazolium (MTT) colorimetric assay. The expression of cyclin E mRNA was measured by RT-PCR. Phosphorylated Akt1(Thr308) and p27 kip1 were analyzed by Western blotting. Results: TNF-α-stimulated proliferation of mesangial cells was inhibited by LXA 4 in a dose-dependent manner. The marked increments in cyclin E mRNA expression induced by TNF-α during proliferation of mesangial cells were down-regulated by LXA 4. Threonine phosphorylated Akt1 proteins at 308 site stimulated by TNF-α was reduced by LXA 4. TNF-α-induced decrements in expression of p27 kip1 proteins was ameliorated by LXA 4 in a dose-dependent manner. Conclusion: TNF-α-induced proliferation of rat mesangial cells can be inhibited by TXA 4 through the mechanism of Akt 1/p27 kip1 pathway-dependent signal transduction.

Antitumor Activity of the Ganoderma Lucidum Spore Alcohol Extract in Vitro

Several cancer cell lines(epithelioma cells or leukemia cells)from human being or mouse were first used to study the antitumor activity of the Ganoderma lucidum spore alcohol extract(GLSAE)in vitro by the MTT test A comparision was made between the sporodermbroken(SB)and sporoderm nonbroken(SN)GLSAE It was showed that both GLSAE SB and GLSAE SN could inhibit the proliferation of these cancer cells,but the activity of GLSAE SB was much higher than that of GLSAE SN These results suggested that Ganoderma lucidum spore could probably be used for tumor treatment

Sustained Contraction of Isolated Rabbit Thoracic Aortic Rings in Endothelial-dependent Manner Induced by βγ-CAT

In vertebrates, non-lens βγ-crystallins are widely expressed in various tissues and their functions are not well known. The molecular mechanisms of trefoil factors （TFFs）, which involved in mucosal healing and tumorigenesis, have remained elusive. βγ-CAT is a novel multifunctional protein complex of non-lens βγ-crystallin and trefoil factor from frog skin secretions. Here we report that βγ-CAT could induce sustained contraction of isolated rabbit aortic rings in dosage （2-35nmol/L） and endothelium dependent manners （P〈0.01 ）. In addition, in situ immunofluorescence indicated that positive TNF-α signals were mainly detected at the endothelial cell layer of βγ-CAT （25nmol/L） treated rings. Furthermore, βγ-CAT induced primary cultured rabbit thoracic aortic endothelial cells （RAECs） rapidly to release TNF-α. After βγ-CAT （25nmol/L） treated for 10 and 30min, the levels of the endothelial cells released TNF-ct were 34.17±5.10 pg/mL and 98.01±4.67 pg/mL （P〈0.01）, respectively. In conclusion, βγ-CAT could induce sustained contraction of isolated aortic rings, and the contractile effect might be partially explained by the release of TNF-α. These findings will give new insight into understanding the functions and physiological roles of non-lens βγ-crystallins and trefoil factors.

The Role of Predominant Expression of Th2 Type Cytokines Gene in the Genesis and Development of Human Gliomas

Objective: To explore the expression of Th1/Th2 cytokines gene in human gliomas and its role in the genesis and development of human gliomas.Methods: Using IL-2 and IFNγ as Th1 type cytokines, IL-4, IL-6 and IL-10 as Th2 type cytokines, the biological activity of cytokines in the supernatant of glioma cell lines was assayed by ELISA method, and the gene expression of Th1/Th2 cytokines in human glioma cells, glioma infiltrating lymphocytes and glioma cell lines were detected by RT-PCR.Results: There was predominant expression of Th2 type cytokines in human glioma cells, glioma infiltrating lymphocytes and glioma cell lines, but there was no such expression in normal brain tissues.Conclusion: It suggested that there is a relationship between the Th2 type cytokines expression in human gliomas and the immunosupressive status of human glioma patients. The predominant expression of Th2 type cytokines may play an important role in the genesis and development of human gliomas. Key words glioma - Th1/Th2 - gene expression - RT-PCR This project was supported by a grant from National Natural Sciences foundation of China (No. 30271335).

Uptake of bacterial lipopolysaccharide and expression of tumor necrosis factor α mRNA in isolated rat intrahepatic bile duct epithelial cells *

AIM To study the uptake of bacterial lipopolysaccharides (LPS) and expression of tumor necrosis factor α mRNA (TNF α mRNA) with cultured rat intrahepatic bile duct epithelial cells.

Arsenic trioxide inhibites transgenic tumor necrosis factor-α promoter activity in vascular smooth muscle cells and THP-1 monocytes in vitro

Aim This study was to evaluate the effect of arsenic trioxide （As2O3） on the transgenic TNF-α promoter activity in cultured vascular smooth muscle cells （VSMCs） and THP-1 monocytes. Methods Human TNF-α promoter was constructed by reporter gene system and was transiently transfected into VSMCs and THP-1 in vitro. The promoter activity was tested by luciferase activity with or without LPS and Ang Ⅱ stimulation, before and after different dosage of As2O3 treatment. Results 1. TNF-α promoter effectively expressed in VSMCs and THP-1 compared with CMV promoter （58.3% and 80.9%, respectively）. Both LPS and Ang Ⅱ significantly up-regulated TNF-α promoter activity （P〈0.05）. 2. As2O3 significantly inhibited, both intact and LPS/Ang Ⅱ stimulated promoter activity, in a dose dependent manner （P〈0.05）, and in both cell type. Conclusion These results manifested that, the inhibition effect of As2O3 on the activity of human TNF-α promoter indicated its potential inhibition on pro-inflammatory cytokine genes expression at transcriptional level and its potential anti-inflammatory property in the cardiovascular system.

The Expression of Apoptosis-Related Genes Bcl-2 and Bax Protein and Apoptosis Positivity in Cervical Carcinoma during Irradiation

To evaluate the apoptosis positivity, the expression of Bcl-2, bax proteinsin 30 patients with squamous cell cervix carcinoma before and after radiotherapy. Methods: By usingimmuno-histochemical and TDT-dUTP nick end labelling techniques, 30 cases of squamous cell cervicalcarcinoma were analyzed. Results: The apoptosis positivity before and after irradiation was 76.7%and 100% respectively, with the difference being significant (P 【 0.05); The positive rates of Bcl-2protein before and after irradiation were 73.3% and 46.7% respectively, with the difference beingsignificant (P 【 0.05); The positive rates of bax protein before and after irradiation were 86% and100% respectively, with the difference being significant (P 【 0.05). Conclusion: bax and Bcl-2protein play an important role in apoptosis induced by fractionated radiation therapy. Apoptosisinduced by irradiation is contributed to upregulation of bax protein or downregulation of Bcl-2protein.

The Effect of As_2O_3 on the Epression of Drugresistance Molecule in Malignant Neoplasmas

Objective: To detect the action of arsenic trioxide (As_2O_3) on theexpression of Tumor drug-resistant molecule. Methods: APL cell line MR_2 resistant to all-transretin-oic acid (ATRA ) was put into research, while APL cell line NB_4 was used for control. Theim-munocytochemical assays were used to detect the expressions of P-glycoprotein (P_(gp)) andGluta-thione S-transferase ( GST) . Results: Not only the expression of P_(gp) in MR_2 cell line(30%-40%) was significantly higher than that in NB4 cell line (10%-20% ) (P < 0.001), but also theexpression of GST in MR_2 cell line (60. 4 +- 4.0 )-( 66.5 +- 4.4) was significantly higher thanthat in NB4 cell line (28.3 +- 5.6)-(31.2 +- 5. l)(P < 0.05). As_2O_3 at the concentration of0.5-2.0 μmol/l could significantly decrease the expression of P_(gp) and GSTπ, but could donothing about the expression of GSTα and GSTμ. Conclusion: The lower expression of P_(gp) andGSTπ might be the sensitive indications of frustrating drug-resistance, while GSTα and GSTμ mightnot be the case. ATRA might be the substrates of P_(gp) transmission and GSTπ catalysis .

Expression of vascular endothelial growth factor and its prognostic significance in gastric carcinoma

AIMS To investigate the clinical significance of vas- cular endothelial growth factor (VEGF) expression in gastric carcinoma. METHODS The expression of VEGF in 128 gastric carcinomas was investigated by immunohistochemical staining with a polyclonal antibody against VEGF. Cor- relations between the expression of VEGF and various clinicopathologic factors and prognosis were studied. RESULTS The VEGF-rich expression rate was 64.1% in gastric carcinoma. VEGF-rich expression rate of patients with stage Ⅲ and stage Ⅳ disease was greater than that of patients with stage f disease (P <0.05). Significant differences of expression rate ex- isted with respect to growth pattern,serosal invasion and lymph node metastasis. The VEGF-rich rate was much higher in tumors with expanding growth pattern (71.8%) or serosal invasion (73.5%) than in those with infiltrative growth pattern (52.0%) or non-serosal invasion (53.3%) (P<0.025,respectively),and it was also significantly higher in patients with lymph node metastases (75.0%) than in those without such metastases (50.0%) (P<0.05). In addition,postop- erative survey of 86 patients who had been followed up for at least 5 years demonstrated that the 5-year sur- vival rate of patients with VEGF-rich tumors was signifi- cantly lower than that of patients with VEGF-poor tu- mors (P<0.05). CONCLUSIONS The expression of VEGF may be as- sociated with the invasion and metastasis and may also be a useful prognostic indicator of gastric carcinoma.

Immunotherapy of EGFR-positive Tumor Based on Recombinant EGFR Phage Vaccine

Objective： To construct the recombined phage which was inserted with the gene of extracellular domain of chicken EGFR as a new type vaccine and to evaluate the efficiency of the phage vaccine against the EGFR positive tumor. Methods： The T7 phage display system was used to display 5 fragments of the extracellular domain of chicken EGFR. The EGFR was expressed as a fused protein on the surface of the T7 phage 10B capsid protein. The EGFR expression of the phage vaccine was verified by the Western blot analysis. The anti-EGFR antibody was detected by ELISA. The splenic lymphocytes of the immunized mice were separated and used to determine the cellular immunotoxic effect against A431 cells. The phage vaccines were injected into the C57 mice 4 times before lewis lung cancer cells were inoculated. The tumor volume was recorded to evaluate the anti-tumor effect of each vaccine. Results： Five phage vaccines inserted with the chicken EGFR gene were constructed successfully. Western blot assay showed that the extracellular domain of chicken EGFR proteins was displayed on the surface of the phage. The specific antibody was induced in the immunized mice compared with the control group. The splenic lymphocytes of the immunized mice were shown to be immunotoxic against A431 cells. The killing rates of the experimental groups were higher than in the control group （P〈0.001, by t test）. The highest killing rate was （45.74±7.21）%. The tumor growth was inhibited in the experimental groups as compared with the control group （P〈0.05 in C1, C2, C3 and C4 groups, P〉0.05 in C5 group）. Conclusion： The phage vaccine could induce both the protective and therapeutic antitumor immunity against EGFR-positive tumor.

Measurement of Serum Total and Free Prostate-Specific Antigen for Breast Cancer Diagnosis in Women

Objective: To investigate the diagnostic value and the relationship between the clinicopathological features and the levels of total and free prostate-specific antigen (PSA) in women with breast cancer.Methods: Using the microparticle enzyme immunoassay system, we measured the concentrations of these markers in the sera of 85 women with breast cancer and in 30 healthy women.Rseults: The lowest detection level for both markers was 0.01 ng/ml. Free PSA levels were significantly higher in women with breast cancer than that in healthy women (P<0.05). The percentage of free PSA predominant subjects was 37.6% in breast cancer patients and 3.3% in healthy women. Cut-off values were 0.36 ng/ml for total PSA and 0.02 ng/ml for free PSA. In women with breast cancer, total PSA positivity was 23.5% and free PSA positivity was 27.1%. Compared to negatives, total PSA positive patients had a higher percentage of lymph node involvement tumours (P>0.05). However, patients with predominant free PSA had a higher percentage of early stage than patients with predominant PSA-ACT.Conclusion: Although the sensitivity of free PSA predominance is low (37.6%) in distinguishing women with breast cancer from healthy women, its specificity is high (97.0%).Free PSA predominance tends to be present in early stage tumours. These findings may indicate clinical significance of preoperative measurement of serum total and free PSA in women with breast cancer.

Molecular mechanism of Radix astragali on improvement of insulin sensitivity of SD rats treated with low dose dexamethasone

Aim To reveal the main active components and the action mechanisms of Radix astragali on insulin sensitivity improvement, we have investigated the effects of polysaccharide portion and saponin portion of Radix astragali extracts on blood biochemical indices and related gene expression of dexamethasone-induced SD rats. Methods SD rats （6 per group） received 2 μg/day subcutaneous dexamethasone for 4 weeks plus same dose （10 g material/kg） of polysaccharide or saponin extracts of Radix astragali. Blood samples, kidney tissues and epididymal fat pads were taken at the end of the experiment. Serum triglyceride （TG）, total cholesterol （TC）, low density lipoprotein cholesterol （LDLC）, high density lipoprotein cholesterol （HDLC）, glucose （GLU） and insulin （INS） levels were measured, respectively, mRNA levels of angiotensinogen in kidney, adiponectin and leptin as well as TNF-α in epididymal fats were determined by RT-PCR assay using GAPDH gene as an internal control. Results Both of polysaccharide and saponin extracts of Radix astragali exhibited positive effects in reducing serum triglycerides, glucose, and insulin levels of dexamethasone-induced SD rats. The saponin group showed more improvements on quantitive insulin sensitivity check index （QUICKI） than the polysaccharide group did. Both of the extracts down-regulated kidney angiotensinogen and fat TNF-α mRNA levels while they were simultaneously up-regulating fat adiponectin and leptin mRNA levels. No significant difference was found between actions of the two extracts. Conclusion Both of polysaccharide and saponin extracts of Radix astragali can improve insulin sensitivity. This action might be closely related to down-regulation of angiotensinogen, TNF-α and up-regulation of adiponectin and leptin expression. The results partly explained the improvement of type Ⅱ diabetes and diabetic nephropathy by Radix astragali. The similar actions of the two crude extracts suggest that unknown key active compounds might exist in both and remain to be discovered.

Over-expressed and truncated midkines promote proliferation of BGC823 cells in vitro and tumor growth in vivo

AIM： To determine whether midkine （M/O and its truncated form （tMK） contribute to gastric tumorigenesis using in vitro and in vivo models. METHODS： Human MK and tMt（ plasmids were constructed and expressed in BGC823 （a gastric adenocarcinoma cell line） to investigate the effect of over-expressed MK or tMK on cell growth and turmorigenesis in nude mice. RESULTS： The growth of MK-transfected or tMK- transfected cells was significantly increased compared with that of the control cells, and tMK-transfected cells grew more rapidly than MK-transfected cells. The number of colony formation of the cells transfected with MK or tMK gene was larger than the control cells. In nude mice injected with MK-transfected or tMK-transfected cells, visible tumor was observed earlier and the tumor tissues were larger in size and weight than in control animals that were injected with cells without the transfection of either genes. CONCLUSION： Over-expressed MK or tMtC can promote human gastric cancer cell growth in vitro and in vivo, and bMK has greater effect than MK. tMK may be a more promising gene therapeutic target compared with MK for treatment of malignant tumors.