SAF-189s, a potent new-generation ROS1 inhibitor, is active against crizotinib-resistant ROS1 mutant-driven tumors.

The ROS1 fusion kinase is an attractive antitumor target. Though with significant clinical efficacy, the well-known first-generation ROS1 inhibitor (ROS1i) crizotinib inevitably developed acquired resistance due to secondary point mutations in the ROS1 kinase. Novel ROS1is effective against mutations conferring secondary crizotinib resistance, especially G2032R, are urgently needed. In the present study, we evaluated the antitumor efficacy of SAF-189s, the new-generation ROS1/ALK inhibitor, against ROS1 fusion wild-type and crizotinib-resistant mutants. We showed that SAF-189s potently inhibited ROS1 kinase and its known acquired clinically resistant mutants, including the highly resistant G2032R mutant. SAF-189s displayed subnanomolar to nanomolar IC50 values against ROS1 wild-type and mutant kinase activity and a selectivity vs. other 288 protein kinases tested. SAF-189s blocked cellular ROS1 signaling, and in turn potently inhibited the cell proliferation in HCC78 cells and BaF3 cells expressing ROS1 fusion wild-type and resistance mutants. In nude mice bearing BaF3/CD74-ROS1 or BaF3/CD74-ROS1G2032R xenografts, oral administration of SAF-189s dose dependently suppressed the growth of both ROS1 wild-type- and G2032R mutant-driven tumors. In a patient-derived xenograft model of SDC4-ROS1 fusion NSCLC, oral administration of SAF-189s (20 mg/kg every day) induced tumor regression and exhibited notable prolonged and durable efficacy. In addition, SAF-189s was more potent than crizotinib and comparable to lorlatinib, the most advanced ROS1i known against the ROS1G2032R. Collectively, these results suggest the promising potential of SAF-189s for the treatment of patients with the ROS1 fusion G2032R mutation who relapse on crizotinib. It is now recruiting both crizotinib-relapsed and naive ROS1-positive NSCLC patients in a multicenter phase II trial (ClinicalTrials.gov Identifier: NCT04237805).

[Establishment of transformation system of Claviceps purpurea Cp-1 strain].

Claviceps pururea Cp-1 strain established in our lab is capable of producing variety of bioactive ergot alkaloids, and is broadly used by the pharmaceutical companies. To engineer the strain genetically for the production of specific ergot alkaloid, an effective transformation system must be set up first. However, the reported transformation system is not suitable for this strain due to different genetic backgrounds and the heterogeneity of Claviceps. Thus, in this paper, the hyphae of Cp-1 strain were used to prepare protoplasts by lywallzyme. The formation of protoplasts was investigated under different concentrations and incubation time of enzyme. The strain was tested for sensitivity to several antibiotics at different concentrations. Finally, the genetic transformation system of Cp-1 strain was established. The results suggest that protoplasts were formed efficiently by using 1% lywallzyme at 25 ℃ for 2 h.Transformants were obtained by PEG mediated protoplast transformation of Cp-1 strain with plasmid pAN7-1,using 1.5 mg·m L(-1) hygromycin B as the selective marker.The exogenous gene in the plasmid pAN7-1 was integrated into the genome of Cp-1 strain transformant as demonstrated by PCR result. This study laid an important foundation for genetic manipulation of Cp-1 strain.

FokI Polymorphism of the Vitamin D Receptor Gene Is Associated with Susceptibility to Gastric Cancer: A Case-Control Study.

Vitamin D is a potential protective agent against cancer, and its activity is mediated mainly by vitamin D receptor (VDR). The FokI polymorphism (rs10735810) represents a T-to-C transition (ATG to ACG) in exon 2 of the VDR gene, and this ATG represents the translation-initiation codon, encoded by the f allele. The FokI polymorphism results in the generation of a protein shortened by three amino acids, translated from the downstream ATG codon (the F allele). We investigated the relationship between the FokI polymorphism and gastric cancer in a Chinese Han population. A total of 187 patients and 212 healthy controls were enrolled. The FokI polymorphism was detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis. The f allele frequency was higher in patients than that in controls (51.6% and 43.6%, P < 0.05). Multivariate logistics regression analysis revealed patients with the f allele (Ff + ff) showed a higher risk of gastric cancer [odds ratio (95% confidence interval) 2.73 (1.13~4.32)]. Patients with the f allele (Ff + ff) also presented a poorly differentiated type of gastric cancer (P < 0.05) and higher levels of C-reactive protein on admission than the FF group (5.5 ± 2.4 mg/L vs. 3.4 ± 1.3 mg/L, P < 0.05). Here, we show an association between the VDR FokI polymorphism and the susceptibility to gastric cancer, which may be helpful for early detection of high-risk individuals with the f allele for gastric cancer. Conversely, the F allele may be a protective factor against gastric cancer.

Prevention of Dexmedetomidine on Postoperative Delirium and Early Postoperative Cognitive Dysfunction in Elderly Patients Undergoing Thoracoscopic Lobectomy.

Objective: To investigate the effect of dexmedetomidine on postoperative cognitive function and delirium in elderly patients undergoing thoracoscopic lobectomy. Methods: 109 elderly patients (age is more than 65 years) who underwent thoracoscopic lobectomy in our hospital from June 2020 to Feb 2022 were randomly divided into the dexmedetomidine (DEX) group (n = 54) and the control group (n = 55). The patients in the experimental group were given dexmedetomidine by intravenous pump, intravenous pump 0.5 µg/kg within 10 minutes, and maintained the speed of 0.5 µg/(kg. h) to 30 min before the operation was ended. The control group was given the same amount of normal saline. Delirium assessment-severity (CAM-S) assessment and Mini-Cog were used to assess the severity levels of POD and POCD 24 h before, 6 hours after, one day after the operation, three days after the operation, and 1 week after the operation. Serum TNF-αα and NSE levels were assessed by using enzyme-linked immunosorbent assay. NRS pain marks were assessed in the DEX group at postanesthesia care unit (PACU) and 24 postoperation. Surgical pierhysmographic index (SPI) evaluation was performed at five time points. Results: The Mini-Cog scores in the DEX group were markedly enhanced compared with those in the saline group 6 and 24 hours after the operation. The SPI values in the DEX group were markedly reduced within 2 min after intubation and at surgical sutures. Moreover, the CAM scores in the DEX group were markedly reduced 24 hours after the operation. Tumor necrosis factor-α (TNF-α) and neuron-specific enolase (NSE) levels were significantly decreased in the DEX group at T1∼T3. Conclusion: The use of dexmedetomidine in the thoracoscopic lobectomy in elderly patients could reduce the occurrence and severity of postoperative cognitive dysfunction and delirium.

Intranasal Immunization with Zika Virus Envelope Domain III-Flagellin Fusion Protein Elicits Systemic and Mucosal Immune Responses and Protection against Subcutaneous and Intravaginal Virus Challenges.

Zika virus (ZIKV) infections in humans are mainly transmitted by the mosquito vectors, but human-to-human sexual transmission is also another important route. Developing a ZIKV mucosal vaccine that can elicit both systemic and mucosal immune responses is of particular interest. In this study, we constructed a recombinant ZIKV envelope DIII (ZDIII) protein genetically fused with Salmonella typhimurium flagellin (FliC-ZDIII) as a novel mucosal antigen for intranasal immunization. The results indicated that the FliC-ZDIII fusion proteins formulated with E. coli heat-labile enterotoxin B subunit (LTIIb-B5) adjuvant greatly increased the ZDIII-specific IgG, IgA, and neutralizing titers in sera, and the ZDIII-specific IgA titers in bronchoalveolar lavage and vaginal fluids. Protective immunity was further assessed by subcutaneous and intravaginal ZIKV challenges. The second-generation FliCΔD3-2ZDIII was shown to result in a reduced titer of anti-FliC IgG antibodies in sera and still retained the same levels of serum IgG, IgA, and neutralizing antibodies and mucosal IgA antibodies without compromising the vaccine antigenicity. Therefore, intranasal immunization with FliCΔD3-2ZDIII fusion proteins formulated with LTIIb-B5 adjuvant elicited the greatest protective immunity against subcutaneous and intravaginal ZIKV challenges. Our findings indicated that the combination of FliCΔD3-2ZDIII fusion proteins and LTIIb-B5 adjuvant for intranasal immunization can be used for developing ZIKV mucosal vaccines.

[Study of the effect of light source stability on the signal to noise ratio in degenerate four wave mixing experiment].

The effects of the stability of dye laser on the signal to noise ratio in degenerate four-wave mixing (DFWM) were first investigated in iodine vapor using forward geometries. Frequency-doubled outputs from a multi-mode Nd : YAG laser pumped dye laser with laser dye PM580 dissolved in ethanol was used. With the help of forward compensated beam-split technique and imaging detecting system, the saturation intensity of DFWM spectrum in the iodine vapor at 5 554.013 nm was first measured to be 290 microJ under the condition of atmospheric pressure and room temperature. The features of the dye laser such as wavelength ranges, beam quality and energy conversion efficiency decreased gradually with increasing pumping service use, pulse number and intensity. Additionally, with the comparison of the stable and unstable dye laser output, it was found that the instability of dye laser output had greatly influenced the DFWM signal and decreased the signal to background noise ratio. Shot to shot jitter and the broadening in the output frequency leads to an effective broadening of the recorded spectrum and loss of the DFWM signal to noise ratio under the same pumping intensity at different time. The study is of importance to the detection of trace atom, molecule and radical in combustion diagnosis.

[Message RNA expression of LUNX, CK19 and CEA genes in NSCLC with micrometastasis in lymph nodes].

OBJECTIVE: To investigate the correlation of mRNA expression level of three cancer-associated genes-LUNX mRNA, CK19 mRNA and CEA mRNA with metastasis in lymph nodes and histopathological staging in non-small cell lung cancer (NSCLC). METHODS: Fifty-six tumor tissue samples and 103 regional lymph node samples were obtained from 56 patients with NSCLC, and another 35 lymph node samples as control from 15 patients with benign pulmonary diseases. The mRNA expression of LUNX, CK19 and CEA genes was detected in these samples by semi-quantitative RT-PCR analysis (reverse transcriptase polymerase chain reaction), meanwhile, all lymph nodes were also examined by conventional pathological method. RESULTS: mRNA expression of LUNX, CK19 and CEA genes in the regional lymph nodes of NSCLC was significantly higher than that in those of benign lung diseases (P < 0.05). Compared with conventional pathological method, RT-PCT was more sensitive (P < 0.05). No significant correlation was found between positive mRNA expression of LUNX mRNA and CK19 mRNA in the lymph nodes and histopathologic type of lung cancer (P > 0.05). But positive expression rate of CEA mRNA in the lymph nodes from adenocarcinoma patients was significantly higher than that in these from squamous cell carcinoma and other types of NSCLC (P < 0.05). The expression level of LUNX mRNA in the lymph nodes was positively correlated with TNM stages. CONCLUSION: LUNX mRNA and CK19 mRNA may serve as a molecular marker for detection of lymph node micrometastasis in patient with non-small cell lung cancer, but LUNX mRNA is superior to CK19 mRNA in both sensitivity and specificity.

[Application of UV similarity in quality control of Chinese medical injection].

OBJECTIVE: To propose an algorithm of the similarity between UV-spectra for controlling the quality of Chinese medical injection. METHOD: The similarity is calculated base on the data of two UV-spectrum curves, thus the sample quality is evaluated according to the average value of the similarity of the UV-spectra between sample with each of standard samples; the proposed algorithm of the similarity is compared with cosine of vectorial angle and relative coefficient; the proposed method for quality evaluation of traditional Chinese injection is discussed with the method according to the chromatographic fingerprint similarity. RESULT: The calculated similarity with the proposed method can be sensitive to show the difference between UV-spectra. This proposed method for quality evaluation was applied for assessing quality of 14 batches of Xiangdan injection and the results well coincident with sample quality. CONCLUSION: The proposed method can be used to evaluate the quality of the Chinese medical injection according to the similarity of two UV-spectrum curves.

Loss of ß-arrestin-2 and Activation of CXCR2 Correlate with Lymph Node Metastasis in Non-small Cell Lung Cancer.

Background : Although ß-arrestin-2 (ß-arr2) and CXCR2 have been shown to affect various malignant tumors, their exact roles in lung cancer remain unclear. We investigated expression of ß-arr2 and CXCR2 in patients with non-small cell lung cancer (NSCLC) and their correlation with lymph node metastasis and prognosis. Methods : We reviewed medical records of 136 patients with NSCLC who underwent surgical resection, and assessed their specimens immunohistochemically for expression of ß-arr2 and CXCR2 in primary tumors and metastatic lymph nodes (MLNs), respectively. Results: High ß-arr2 expression was seen in 63 specimens (46.3%), and was significantly associated with male patients (P=0.011), squamous cell carcinoma (P=0.003), and lymph node metastasis (P<0.001). High CXCR2 expression was seen in 62 specimens (45.6%), and was significantly correlated only with lymph node metastasis (P<0.001). Expression of ß-arr2 was significantly lower at MLNs than at primary lesions (Z=-2.315; P=0.021; Wilcoxon signed-rank), whereas CXCR2 expression was significantly higher in MLNs than in primary lesions (Z=-3.712; P<0.001; Wilcoxon signed-rank). No relationship was seen between ß-arr2 and CXCR2 expression in primary lesions (r=-0.065, P=0.548; Spearman rank coefficient), but they were inversely related in MLNs (r=-0.263, P=0.012). Kaplan-Meier survival curve was shown that low ß-arr2 and high CXCR2 expressions was associated with poor survival (log-rank: χ2=5.926, P=0.015). Conclusions : ß-arr2 may promote lymph node metastasis in NSCLC by modulating CXCR2 activation.

MiRNA-155 mediates TAM resistance by modulating SOCS6-STAT3 signalling pathway in breast cancer.

Breast cancer is the second leading cause of cancer induced death in women. Tamoxifen is an endocrine therapy which is administered to 70% of all breast cancer patients with estrogen receptor alpha (ERα) expression. Despite the initial response, most patients eventually acquire resistance to the drug. MicroRNAs (miRNAs) are a class of small non-coding RNAs which have the ability to post-transcriptionally regulate gene expression. Although the role of a few miRNAs has been described in tamoxifen resistance, little is known about how concerted actions of miRNAs targeting biological networks contribute to its resistance. In this study, we identified that miR-155 is frequently up-regulated in breast cancer with tamoxifen resistance. Ectopic expression of miR-155 induces cell survival and resistance to TAM, whereas inhibition of miR-155 causes cells to apoptosis and enhances TAM sensitivity. Further, we identified SOCS6 as a new direct target of miR-155. Sustained overexpression of miR-155 resulted in repression of SOCS6 protein and mRNA levels, and knockdown of miR-155 increased SOCS6 expression. Introduction of SOCS6 cDNA lacking the 3'-UTR abrogated miR-155-induced cell survival and chemoresistance. Finally, it was verified that SOCS6 or inhibition of STAT3 could inhibit miR-155 STAT3 activation and cell proliferation. In conclusion, our study reveals a molecular link between miR-155 and SOCS6-STAT3 and presents an evidence that miR-155 is a critical therapeutic target in breast cancer.

TRPM7 promotes the metastatic process in human nasopharyngeal carcinoma.

Our study observed the relationship between transient receptor potential melastatin 7 (TRPM7) expression and the metastatic process of nasopharyngeal carcinoma (NPC). We found that TRPM7 was overexpressed in 102 out of 206 (49.5%) human NPC cases and was significantly associated with clinical stage and lymphatic and distant metastasis. The results suggested that TRPM7 promotes NPC cell migration and invasion in vitro. Further, TRPM7 was correlated with poor clinical outcome and was an independent predictor for 5-year overall survival rate (HR, 1.832; 95% CI, 1.237-4.146 [P = 0.041]). In conclusion, TRPM7 promotes the metastasis of NPC and may serve as a prognostic marker in NPC patients.

Effect of different iodine intake on schoolchildren's thyroid diseases and intelligence in rural areas.

BACKGROUND: Reports are increasingly appearing on the side effects caused by excessive iodine intake. Our objective was to find out whether iodine excess would impair the thyroid function and intelligence of schoolchildren in rural areas of China. METHODS: A comparative epidemiological study was made on thyroid function and intelligence of the schoolchildren in the areas of low, moderate or excessive intake of iodine. In the area of low intake of iodine (Panshan, Liaoning province, median urinary iodine (MUI) was 99 microg/L), of moderate intake of iodine (Zhangwu, Liaoning Province, MUI was 338 microg/L) and of excessive intake of iodine (Huanghua, Hebei Province, MUI was 631 microg/L). The numbers of schoolchildren from each area selected to take part in a Chinese version of Raven's Test were 190, 236 and 313, respectively, and then 116, 110 and 112 of them were tested for thyroid function, thyroid autoantibody (TAA) and urinary iodine (UI). RESULTS: There were no significant differences in the incidences of overt hyperthyroidism, subclinical hyperthyroidism and overt hypothyroidism in Panshan, Zhangwu and Huanghua. But significant differences were found in the incidences of subclinical hypothyroidism (P = 0.001) in these three areas. The incidences of subclinical hypothyroidism in Huanghua and Zhangwu were 4.76 and 3.37 times higher than that in Panshan. TAA were negative in all the schoolchildren with subclinical hypothyroidism except for one. No significant difference was found among the rates of thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TGAb) in these three areas. Mean serum thyroglobulin (TG) value of Huanghua was markedly higher than those of the other two (P = 0.02). Mean serum TG value of Zhangwu was higher than that of Panshan but the difference was not significant. Mean IQ value of the schoolchildren in Huanghua was markedly higher than that for Zhangwu (P = 0.001). Mean IQ value of the schoolchildren in Panshan was lower than that of Huanghua and higher than that of Zhangwu but, again, the differences were not significant. CONCLUSIONS: The increase of iodine intake may increase the risk for schoolchildren of subclinical hypothyroidism. In the area of iodine excess, most of the subclinical hypothyroidism cases are not of autoimmune origin. No obvious effect of excess iodine was found on mental development of schoolchildren.

[Correlation between serum thyroglobulin and thyroid stimulating hormone in populations with non-toxic goiter].

OBJECTIVE: To investigate the correlation between serum thyroglobulin (Tg) and thyroid stimulating hormone (TSH) in populations with non-toxic goiter. METHODS: Thyroid ultrasonography was conducted, and determination of serum triiodothyronine (FT3), thyroxin (FT4), Tg, thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TGAb) and TSH, and urine iodine were carried out among 609 persons with an average age of 35.4 in Panshan county, a mild iodine-deficient area in Liaoning province, and 1136 persons aged 39.1 on average in Zhangwu county, an iodine sufficient area in Liaoning province, totally 2 320 persons. RESULTS: The serum TSH levels of the populations with diffuse and nodular goiter in Panshan county were (0.11 +/- 0.08) mU/L and (0.90 +/- 0.10) mU/L, significantly higher than those of the normal population in the same areas (1.33 +/- 0.04) mU/L, both P < 0.01. The serum TSH levels of the populations with diffuse and nodular goiter in Zhangwu county were (1.29 +/- 0.13) mU/L and (0.89 +/- 0.14) mU/L, significantly higher than those of the normal population in the same areas (1.74 +/- 0.04) mU/L, both P < 0.01. The serum TSH level of normal population in iodine deficient area was significantly lower than that of normal population in iodine sufficient area (P < 0.01). The serum TSH level of the normal population in Panshan County was significantly lower than that of the normal population in Zhangwu County (P < 0.01). The serum Tg level of the populations with diffuse and nodular goiter in Panshan county were (12.0 +/- 1.21) ng/L and (50.1 +/- 12.20) ng/L, significantly higher than that of the normal population in the same area (9.1 +/- 2.44) ng/L, P < 0.05 and P < 0.01. The serum Tg level of the populations with diffuse and nodular goiter in Zhangwu county were (12.4 +/- 2.11) ng/L and (61.2 +/- 11.86) ng/L, significantly higher than that of the normal population in the same area (7.7 +/- 2.3) ng/L, P < 0.05 and P < 0.01. The Tg level of the normal population in Panshan County was significantly higher than that of the normal population in Zhangwu county (P < 0.01). The Tg level was positively correlated with the thyroid volume in the nodular goiter population. CONCLUSION: The occurrence of goiter is not directly related to TSH stimulation in mild iodine deficient area. Nontoxic goiter shows a thyroid autonomic function. The major cause of high serum Tg in nodular goiter may be the leakage of Tg into blood circulation caused by degeneration of large colloid follicles and destruction of follicular architecture, not the volume increase of thyroid and its autonomic function.

[Multifactor analysis of relationship between the biological exposure to iodine and hypothyroidism].

OBJECTIVE: To assess the relationship between the biological exposure to iodine and hypothyroidism. METHODS: Logistic regression model was used to analyze the risk factors of hypothyroidism, according to the epidemiologic data of 3761 adults in 3 kinds of rural communities: mild iodine deficiency area (4 natural villages in Panshan County, Liaoning Province), more than adequate iodine (7 natural villages of Zhangwu County, Liaoning Province), and excessive iodine area (2 natural villages of Huanghua City, Hebei Province). RESULTS: More than adequate iodine and excessive iodine were independent risk factors of subclinical hypothyroidism (OR = 3.172 and 6.391, P < 0.05) and overt hypothyroidism (OR = 3.696 and 9.213, P < 0.05). When interactions of iodine exposure and thyroid peroxidase antibody (TPOAb) or thyroglobulin antibody (TgAb) were included, more than adequate iodine was still a risk factor of subclinical hypothyroidism (OR = 2.788, P < 0.01), but had no such effect on overt hypothyroidism. Interaction of more than adequate iodine and positive TgAb significantly affected subclinical hypothyroidism and overt hypothyroidism (OR = 2.656 and 3.347, P < 0.05). CONCLUSION: More than adequate and excessive iodine exposure are independent risk factors of hypothyroidism. The risk of hypothyroidism grows up and thyroid dysfunction becomes more serious with the increasing of the biological exposure to iodine.

[An epidemiological study on the relationship between selenium and thyroid function in areas with different iodine intake].

OBJECTIVE: To investigate the relationship between selenium status and thyroid dysfunction in 3 areas with different iodine intake. METHODS: An epidemiological research was performed in the rural communities of Panshan County (iodine-deficient area) and Zhangwu County (iodine-sufficient area), Liaoning Province, and Huanghua County, Hebei Province (iodine-excessive area). Serum selenium, TSH, FT3 and FT4 levels were examined in 329 patients with thyroid dysfunction (including clinical hypothyroidism, subclinical hypothyroidism, clinical hyperthyroidism and subclinical hyperthyroidism) and 183 normal inhabitants. RESULTS: The median serum selenium concentrations in Panshan, Zhangwu and Huanghua were 91.4, 89.1, and 83.2 microg/L respectively. There was no difference in serum selenium levels between the patients with subclinical hypothyroidism, clinical hypothyroidism, and clinical hyperthyroidism and their normal controls. The median serum selenium concentration of the subclinical hyperthyroidism patients was 82.6 microg/L, significantly lower than that of the normal controls (87.3 microg/L). The FT3/FT4 ratio was decreased, the FT4 level was increased in the subclinical hyperthyroidism patients in comparison with the normal controls, and no significant difference in FT3 level was found between them. No significant effect of sex and age was found on serum selenium level of normal inhabitants. In normal controls serum selenium was inversely correlated with serum TSH level, and the subjects with serum selenium < or = 80 microg/L had the median TSH level of 2.10 mU/L, markedly higher than that of the subjects with the serum selenium of 80-100 microg/L (1.29 mU/L) and that of the subjects with the serum selenium of 100 approximately 120 micro g/L (1.28 mU/L). For the thyroid dysfunction patients with positive thyroid auto-antibody (TPOAb) in Zhangwu County, the serum selenium was negatively associated with TPOAb level. The serum selenium level of the TPOAb highly positive group (TPOAb > 600 IU/ml) was 83.6 IU/ml, significantly lower than those of the TPOAb lowly positive group and TPOAb moderately positive group (83.6, 92.9 and 95.6 microg/L respectively). CONCLUSION: No obvious effect of selenium status is found on the development of thyroid dysfunction in these three areas. But selenium deficiency can impair thyroid function by means of disturbing thyroid hormone metabolism and decreasing antioxidant ability of the thyroid.

[Anti-nasopharyngeal carcinoma effect in vivo and in vitro of Avastin and adenovirus-thymidine kinase suicide gene].

OBJECTIVE: To evaluate the effect of Avastin and adenovirus-thymidine kinase/ Ganciclovir (Ad-TK/GCV) suicide gene system on nasopharyngeal carcinoma in vivo and in vitro. METHODS: The expression of vascular endothelial growth factor (VEGF) by CNE1 cell line was detected by VEGF ELISA. The effect of Avastin and Ad-TK/GCV on CNE1 cell was detected by methyl thiazolyl tetrazolium (MTT) assay. Flow cytometry analysis and Hoechst 33342 staining were adopted to explore the killing mechanism of Ad-TK/GCV. The nude mice models of CNE1 cell xenografts were established. After intra-tumoral injection of PBS, Avastin, Ad-TK/GCV, Ad-Lac-Z/GCV and Ad-TK/GCV + Avastin, tumor volume was measured and tumor inhibitory rate was calculated. Then the tumors were removed and subjected to histological examination. RESULTS: CNE1 cells could produce VEGF. Avastin had no direct effect on CNE1 cells. The killing effect of Ad-TK/GCV increased with the increase of Ad-TK multiple of infection and the prodrug concentration, which was enhanced by the existence of bystander effect. Compared with control group, the death cell rate (P = 0.000) and apoptosis cell rate (P = 0.000) had significant difference. The study in vivo showed the tumors treated with Avastin, Ad-TK/GCV and Ad-TK/GCV + Avastin grew slowly compared with control. Tumor volume of treated groups was significantly smaller than that of control (all P < 0.05 or P = 0.000). Tumor weight of treated groups was significantly lower than that of control (all P = 0.000). The histological examination showed local necrosis in Ad-TK/GCV group and Ad-TK/GCV + Avastin group, poor angiogenesis in Avastin group and Ad-TK/GCV + Avastin group. CONCLUSIONS: Avastin had no direct effect on CNE1 cells in vitro. Ad-TK/GCV suicide gene system killed NPC cells by inducing cell necrosis and apoptosis, which could be enhanced by the existence of bystander effect. Avastin and Ad-TK/GCV suicide gene system could inhibit the growth of NPC CNE1 cell xenografts. Combination therapy had a synergic effect.

[Predictive value of drug resistance-related genes expression in neoadjuvant chemotherapy in patients with non-small cell lung cancer of stage III].

BACKGROUND & OBJECTIVE: P-glycoprotein (P-gp), multidrug resistance-associated protein (MRP), and lung resistant protein (LRP) play important roles in multidrug resistance (MDR). This study was to determine P-gp, MRP, and LRP expression in patients with non-small cell lung cancer (NSCLC) of stage III, and evaluate their predictive value in neoadjuvant chemotherapy. METHODS: Immunohistochemical analyses were performed on 31 patients with NSCLC of stage III before, and after neoadjuvant chemotherapy. RESULTS: The frequency of P-gp, MRP, and LRP expression were 29.0% (9/31), 45.2% (14/31), and 38.7% (12/31) before chemotherapy, and were 61.3% (19/31), 51.6% (16/31), and 41.9% (13/31) after chemotherapy. Of 31 patients, 10 (10/31, 32.3%) expressed both MRP and LRP before chemotherapy, which indicated significant positive correlation between MRP and LRP expression (r=0.061, P< 0.001). In patients with P-gp, MRP, or LRP expression before chemotherapy, the response rates towards chemotherapy were 44.4% (4/9), 28.6% (4/14), and 16.7% (2/12). Of 10 patients with both MRP and LRP expression, only 1 (1/10, 10.0%) responded to chemotherapy. The median survival time of patients who responded to chemotherapy was 31 months, while that of patients who did not responded to chemotherapy was 15 months, that of patients who didn't receive neoadjuvant chemotherapy before surgery was 18 months. CONCLUSION: Patients with both MRP and LRP expression are probably resistant to chemotherapy, the value of neoadjuvant chemotherapy is limit in such patients.