OBJECTIVETo construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.

METHODSLentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.

RESULTSThe recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).

CONCLUSIONVHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Apoptosis ; Carcinoma, Renal Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

Objective To construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines. Methods Lentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by LipofectamineTM 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry. Results The recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05). Conclusion VHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Objective To construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines. Methods Lentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by LipofectamineTM 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry. Results The recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05). Conclusion VHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Objective　To investigate the effects of trace elements on the metabolism of extracellular matrix and explore the physiological and pathological mechanism of trauma. Methods　Based on the experimental and clinical data, it was studied that the action of trace elements in the metabolism of extracellular matrix in trauma repairing. Results　During wound healing, the trace elements were the components of many kinds of enzymes, carriers and proteins. They took part in the synthesis of hormones and vitamins as well as the transmission of information system. They activated many different kinds of enzymes and regulate the levels of free radicals. The trace elements had the complicated effects on the synthesis, decompose, deposition and reconstruction of collagen and other extracellular matrix. Conclusion　The trace elements play an important role in regulating the metabolism of extracellular matrix.

Ojective To evaluate the effect of the collagen injection for skin defect correction. Methods The various kinds of skin defect were injected by medical cosmetic collagen. Results 103 cases of skin defect were treated by collagen injection. Short term therapentic effect inclnding "excellent" and "good" was 85.5%. no complication occurred. Conclusion It showed that the method of collagen injection for skin defect correction is safe, simply and effective.

Objective To determine the changes of elements in scars and study the pathological mechanism of hypertrophic scar . Methods The Ca, Mg, Zn, Fe and Cu contents of blood and normal skin in patients without scar (group A, n = 12), of blood, normal skin and flat scar (group B, n = 8) and of blood, normal skin and hypertrophic scar (group C, n = 12) were determined with a GGX-5 model of atomic absorption spectrophotometry. Results ① There were no differences between the contents of the five elements of blood and normal skin in all patients. ② In group B , the contents of the five elements of flat scar are the same as that in normal skin of the same patient . ③ In group C , the contents of the five elements of hypertrophic scar were lower than that of the normal skin of the same patient . Conclusion The cause of the changes of element contents in hypertrophic scar lies in local wounded tissues instead of the whole body of the patient.

Objective To determine the changes of elements in scars and study the pathological mechanism of hypertrophic scar . Methods The Ca, Mg, Zn, Fe and Cu contents of blood and normal skin in patients without scar (group A, n = 12), of blood, normal skin and flat scar (group B, n = 8) and of blood, normal skin and hypertrophic scar (group C, n = 12) were determined with a GGX-5 model of atomic absorption spectrophotometry. Results ① There were no differences between the contents of the five elements of blood and normal skin in all patients. ② In group B , the contents of the five elements of flat scar are the same as that in normal skin of the same patient . ③ In group C , the contents of the five elements of hypertrophic scar were lower than that of the normal skin of the same patient . Conclusion The cause of the changes of element contents in hypertrophic scar lies in local wounded tissues instead of the whole body of the patient.