AIM: To investigate the possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor (rhbFGF). METHOD: Growth of the cells on blood fibrin clot was studied by phase-contrast, scanning and transmission electron microscopy and by Giemsa stain and MTT assay. RESULTS: The optimal concentration of rhbFGF for proliferation and survival of the cells was 100 ng/mL. The cells also grew on blood fibrin clot scaffold in the low-serum medium containing 100 ng/mL rhbFGF, and a greater number of the cells survived after 48 hours incubation compared to that after 24 hours. The elongated filopodia appeared to bridge the gaps among the fibroblasts after 24 hours incubation. Further incubation to 72 hours, a greater number of platycytes were found to be joined together by lamellopodia. CONCLUSION: 3T3 fibroblasts could grow and survive on blood fibrin clot in the low-serum medium containing rhbFGF, and a combination of blood fibrin clot and rhbFGF may have over proportional effects on wound healing.

Objective To determine whether the antiserum produced by immunizing mice with conotoxin GI coupled with bovine serum albumin (BSA) could neutralize GI conotoxin.Methods The GI-BSA was prepared by glutaraldehyde-coupled method,and the mice were immunized with the GI-BSA to produce antiserum.The antibody neutralization assay was used to test the detoxication of the antiserum.Results The SDS-PAGE protein electrophoresis showed that the coupling reaction of GI hapten with BSA was successful.The two distinct protein bands of GI-BSA were more than 120×103.Each mouse was immunized four times with 99 μg every two weeks.After the fourth immunization,the serum neutralization titer was more than 1:64 000.After the intraperitoneal injection of the mixture of 100 or 200 μl of the antiserum and different doses of GI,75% of the mice survived in the group with 100 μl of the antiserum and 1× LD50 GI(16.3 μg/kg).The same percentage of mice also survived in the group of with 200 μl of serum and 25.8 μg/kg of GI.Conclusion The antiserum produced by immunizing mice with GI-BSA exhibits significant detoxication activity to conotoxin GI.

Objective to investigate the role of Pim‐3 and NF‐κB in the development and progression of infiltrating ductal carcinoma of the breast .Methods Here ,we used immunohistochemistry to detect expression of Pim‐3 and NF‐κB in 75 samples of infiltrating ductal breast carcinoma ,21 samples of intraductal breast carcinoma and 30 normal breast tissues .The relationship of their expression ,as well as their correlation with clinicopathological features and patient survival were assessed .Results In con‐trast ,both Pim‐3 and NF‐κB were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue .In the infiltrating ductal carcinoma ,the positive expression rate of Pim‐3 was 77 .3% ,and that of NF‐κB was 68 .0% ;in duc‐tal carcinoma of the breast ,the positive expression rate of Pim‐3 was 52 .4% ,and that of NF‐κB was 42 .9% ;in the normal breast tissue ,the positive expression rate of Pim‐3 was 23 .3% ,and that of NF‐κB was 16 .7% ;the positive expression rate of Pim‐3 was correlated with tumor size ,histological grade ,and clinicopathological stage ;and that of NF‐κB was correlated with tumor size ,histo‐logical grade ,lymph node metastasis of breast cancer .Spearman rank correlation analysis revealed a positive correlation between Pim‐3 expression and NF‐κB expression in infiltrating breast cancer (r=0 .243) .Conclusion Our results demonstrate that Pim‐3 and NF‐κB play a role in the initiation and development of breast cancer ,thus ,these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer .

AIM: To investigate the possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor (rhbFGF). METHOD: Growth of the cells on blood fibrin clot was studied by phase-contrast, scanning and transmission electron microscopy and by Giemsa stain and MTT assay. RESULTS: The optimal concentration of rhbFGF for proliferation and survival of the cells was 100 ng/mL. The cells also grew on blood fibrin clot scaffold in the low-serum medium containing 100 ng/mL rhbFGF, and a greater number of the cells survived after 48 hours incubation compared to that after 24 hours. The elongated filopodia appeared to bridge the gaps among the fibroblasts after 24 hours incubation. Further incubation to 72 hours, a greater number of platycytes were found to be joined together by lamellopodia. CONCLUSION: 3T3 fibroblasts could grow and survive on blood fibrin clot in the low-serum medium containing rhbFGF, and a combination of blood fibrin clot and rhbFGF may have over proportional effects on wound healing.

Aim To further study the molecular mecha-nism of the herbs with hot nature on the regulational action on TRPV1 channel based on the 7900 Real-time PCR instrument. Methods 7900 PCR instrument was applied to detect the intracellular flurescence of TRPV1 channel in the dorsal root ganglions ( DRG ) neurons and the effect on the TRPV1 ’ s thermo-sensational functions of the selected 11 ingredients from hot herbs was explored. Results TRPV1 channels could be ac-tivated by gradually elevated temperature; the activa-tion process could be blocked by the TRPV1 specific blocking agent capsaizepine. Most of the ingredients from hot-nature herbs had the potential to up-regualate TRPV1 channel function. Conclusions The estab-lished TRPV1 channel detection system based on PCR instrument is suitable for the analysis of regulational functions of drugs on the heat-activated TRPV1 chan-nel;the functions of hot herbs may be related to the up-regualtional effects of its active ingredients on the TRPV1 channel which will further up-regulate energy metabolism.

Aim To investigate the anticancer effect of a new xanthono-pyridine derivative N, N '-( 7-oxo-7H-chromeno[3,2-h] quinoline-5,9-diyl)-bis(2-( pyrroli-din-1-yl)acetamide) (XP-16) on human lung carcino-ma cell line A549 and the potential mechanism. Meth-ods Antiproliferative effect of XP-16 on A549 cells was evaluated by MTT assay, morphological examina-tion and colonial assay. Apoptosis detection was car-ried out using Hoechst 33258 and PI double-dyeing method. Intracellular Ca2+ concentration ( [ Ca2+] i ) and mitochondria membrane potential were detected by fluorospectrophotometer. A549 cells treated with XP-16 were collected for Bad and metallothionein 1 A ( MT-1 A ) transcript analysis by real-time reverse tran-scriptase-polymerase chain reaction ( qRT-PCR) . Re-sults XP-16 inhibited A549 cell proliferation in dose-and time-dependent manner. Typical apoptotic mor-
phology such as chromatin aggregation and nuclear fragmentation was observed in A549 cells treated with XP-16 for 24 h, and the apoptosis was showed in a dose-dependent manner. After treated with XP-16, [ Ca2+] i and mitochondria membrane potential of A549 cells were decreased, and relative mRNA level of Bad and MT-1A was up-regulated. Conclusions XP-16 has anticancer effect on A549 cells through apoptosis, which might be associated with decreasing intracellular Ca2+ concentration and mitochondria membrane poten-tial. Up-regulation of MT-1A expression might be the result of decreased [ Ca2+] i .

AIM: To investigate the antitumor and immunological activities of the polysaccharides obtained from rabdosia rubescens (RRP). METHODS: In vitro and in vivo tumor inhibitory experiments as well as MTT method were used. RESULTS: With increasing concentration, the RRP exerted direct cytotoxicity effect in vitro on the cell growth of EAC mice,and its inhibitory rate in vivo against sarcoma 180 reached 35% ( P

AIM: To investigate the behaviour of 3T3 fibroblast and macrophage co-culture on blood fibrin clot or adipose tissue with recombinant basic fibroblast growth factor (rhbFGF). METHODS: MTT method, inverted contrast microscopy, Giemsa staining as well as scanning electron microscope were used in the present study. RESULTS: The effect of rhbFGF on co-culture of 3T3 fibroblast and mouse macrophage on blood fibrin clot in low-serum DMEM with rhbFGF were monitored, and 3T3 fibroblast and macrophage growed well on the blood fibrin blot in low-serum DMEM with rhbFGF. CONCLUSION: The blood fibrin clot, with its low immunogenicity, could be used as a bionic support for three-dimensional tissue culture, and also a physiological carrier to distribute the growth factor rhbFGF for the cells.

Objective:To explore the identification value of carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC-Ag), cytokeratin 19 fragment (Cyfra211), thyroglobulin (TG), ferritin (Fer) and procalcitonin (PCT) in fine needle aspiration eluent in benign and malignant cervical nodules, and acquire the optimal diagnostic model.Methods:Three hundred and ninety-six single cervical nodule patients who underwent fine needle aspiration biopsy from August 2017 to August 2019 in the Center Hospital of Xiaogan City of Hubei Province were selected. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG, Fer and PCT were detected by electrogenerated chemiluminescence method. The results of cytopathological diagnosis were regard as "gold standard", and the diagnostic efficiency of single and combined indexes in fine needle aspiration eluent were analyzed by receiver operating characteristic (ROC) curve.Results:Among the 396 patients, malignant nodules was in 101 cases, and benign nodules was in 295 cases. The fine needle aspiration eluent levels of CEA, SCC-Ag, Cyfra211, TG and Fer in patients with malignant nodules were significantly higher than those in patients with benign nodules: (27.73 ± 10.63) μg/L vs. (16.81 ± 8.18) μg/L, (1.59 ± 0.74) μg/L vs. (1.09 ± 0.83) μg/L, (3.31 ± 1.48) μg/L vs. (1.66 ± 0.59) μg/L, (144.96 ± 38.93) μg/L vs. (95.03 ± 47.23) μg/L and (191.18 ± 80.13) μg/L vs. (137.87 ± 63.22) μg/L, the PCT was significantly lower than that in patients with benign nodules: (0.61 ± 0.24) μg/L vs. (1.01 ± 0.52) μg/L, and there were statistical differences ( P<0.01). The ROC curve analysis result showed that the CEA, Cyfra211 and TG had super diagnostic value (area under curve>0.7, Youden index>0.5); the area under curve of CEA, Cyfra211 combined with TG was significantly higher than other combined detection of 2 indexes ( P<0.05). Conclusions:The combined detection of CEA, Cyfra211 and TG in fine needle aspiration eluent can effectively distinguish the benign and malignant cervical nodules.

Aim The TRPV1 plasmid was transiently transfected into human embryonic kidney HEK 293T cells to establish the heterologous expression system of TRPV1-channel. Methods The transfection efficiency was confirmed under fluorescence mi-croscope and the TRPV1 protein expression was identified by u-sing Western blot, and the functional characteristics of the chan-nel were studied by using the method of confocal microscopy. Results The transfection rate could reach 40% ~50%; the transfected cells were found to have a clear band at the corre-sponding position that TRPV1 should be, which indicated that TRPV1 channel protein was expressed in the transfected cells. The confocal microscopy imaging result showed that the trans-fected HEK 293T cells were activated by TRPV1 channel ago-nist. Conclusion Transient transfection of HEK 293T cells with TRPV1 channel is successfully constructed and the heterol-ogous TRPV1 channel is verified to have normal calcium-media-ting function.