Renal cell carcinoma (RCC) is a common lethal urological cancer,the distant metastasis of which is the leading cause of death.Although targeted agents have remarkably improved the overall prognosis of RCC patients,nearly all the patients eventually acquire therapeutic resistance.With the advent of immune checkpoint inhibitors,immunotherapy based on tumor microenvironment (TME) has shown a broad scope in clinical application.The deepening understanding of TME leads to the changes of therapeutic strategies for advanced RCC,and the combination of targeted therapy and immunotherapy is exhibiting a promising prospect.Herein,we reviewed the TME characteristics,candidate predictive biomarkers,and possible targets for future development of drugs against RCC.
Carcinoma, Renal Cell/therapy* ; Female ; Humans ; Immune Checkpoint Inhibitors ; Immunotherapy ; Kidney Neoplasms/therapy* ; Male ; Tumor Microenvironment

To study the effect of HBx gene on the apoptosis of the cell lines (L02, HepG2) and the interaction between HBx and X-linked inhibitor of apoptosis protein (XIAP), the apoptosis of pcDNA3.1-HBx transiently transfected cell lines (L02, HepG2) was detected by flow cytometry and the mRNA expression of XIAP was assayed by real-time RT-PCR. Our study showed (1) the morphology of L02/pcDNA3.1-HBx was changed and the appearance of the cells mimicked that of HepG2 cells; (2) HBx gene could be detected in L02/pcDNA3.1-HBx and HepG2/ pcDNA3.1-HBx; (3) the apoptosis rate of L02/pcDNA 3.1-HBx was higher than that of L02 cells (P<0.01) and the apoptosis rate of HepG2/pcDNA3.1-HBx was lower than that of HepG2 cells (P<0.05); (4) the XIAP expression in L02 was about 3 times that in L02/pcDNA3.1-HBx cells (P<0.01), and the expression of XIAP in HepG2/pcDNA3.1-HBx was about 4 times that in HepG2 (P<0.01). It is concluded that HBx gene may promote the apoptosis of normal hepatocytes and inhibit the apoptosis of cells of hepatic carcinoma by regulating the expression of XIAP.

BackgroundHypoxia-inducible factor-1 α (HIF-1α) specific double-stranded RNA ( dsRNA ) mediated by liposome inhibit reinal neovascularization in mice at dose-dependent manner. ObjectiveThe present study was to investigate the inhibitory effect of dsRNA targeting HIF-1α on retinal neovascularization in mice.MethodsModels of oxygen-induced retinal neovascularization were set up in C57BL/6J mouse through exposure of postnatal day 7 ( P7 ) to (75±3) ％ oxygen for 5 days.Fluorescein conjugated Dextran angiography of retinal vascular was performed to identify the retinal neovascularization.The 8 mice of the normal group were raised in the room air.Fifty-one P7 mice exposed to(75±3)％ oxygen for 5 days and then returned to the room air and assigned to control group ( 3 mice),empty vector group( 3 mice) and gene therapy group (45 mice),and the latter were medially divided to 9 groups randomly according to dose-ratio ( liposomes ∶ plasmid).The pSilencer 2.1-U6 hygro was injected in the model mice of empty vector group,and different dose-ratios of pSilencer2.1-U6 hygro-HIF-1α dsRNA were injected respectively in the model mice of various gene therapy groups.Fluorescein conjugated Dextran angiography of retinal vascular was performed to observe the morphology of new blood vessels,and retinal slides were prepared to score the numbers of nuclei extending beyond the inner limiting membrane( ILM ),and expression of vascular endothelial growth factor(VEGF) was detected in the retina by immunohistochemistry.Results The retinal blood vessels of the normal group formed a fined radial branching pattern.The retinal vascular patterns in the control group and the empty vector group were characterized by decreased central perfusion in both the superficial and the deep layers.The abundant vessels were distorted and irregular in the control group and empty vector group,and the obstructed capillary and lots of neovascular tufts were seen.The retinal neovascularization and non-perfusion distraction in the every gene therapy group were reduced markedly with the most severe appearance in 1 ∶ 1 ( liposomes ∶ plasmid) dose-ratio group.Few vascular endothelial cell nucleus extending beyond the ILM were found in the normal group;while a large number of vascular endothelial cell nucleus were showed in the control group and empty vector group with the occurring rate 100％.Statistically,no significant difference was seen in the number of nuclei extending beyond the ILM between the control group and the empty vector group(11.57±5.85 vs 11.53±6.15),however,that in 1∶1 (liposomes∶plasmid) group was reduced markedly ( 2.17 ± 4.23 ) ( P ＜ 0.01 ).Immunohistochemistry revealed that VEGF was faintly expressed in the normal group but strongly expressed in the control group and the gene therapy group.VEGF expressions of various gene therapy groups were weaker than ones of the control group and the empty vector group.ConclusionsRetinal neovascularization can be efficiently inhibited by intravitreal injection of the pSilencer2.1-U6 hygro-HIF-1α dsRNA mediated by liposome.Proportion of 1 ∶ 1 (liposomes ∶ plasmid)has a maximized efficiency.

Objective To summarize the experience and lesson of surgical treatment of acute traumatic brain injury.Methods A total of 516 cases with acute traumatic brain injury treated surgi- cally from January 2001 to December 2004 in our hospital were analyzed retrospectively.Of all,there were 56 cases with simple comminuted depressed fractures,138 with brain contusion and laceration and/ or intracerebral hematoma,122 with epidural hematoma,126 with suhdural hematoma,48 with diffuse brain swelling,18 with open brain injuries and eight with other kind of injury,all of whom were treated with standard large eraniectomy under general anaesthesia.The treatment results were evaluated by Glas- cow Outcome Scale(GOS).Results Of all,standard large craniectomy was performed in 194 cases, of which 304 cases(58.9%)were with good recovery,66(12.8%)with moderate deficit,72(13.9%) with severe deficit,12(2.3%)under persistent vegetative status and 62(12.0%)died 3-6 months after surgery.Conclusions More attention should be paid to surgery for traumatic brain injury.Individual surgical treatment should be performed under surgical principles.

Objective To investigate the effects of desflurane on the delayed rectifier potassium current (Ik ) in acutely dissociated rat parietal cortical neurons. Methods Wistar rats between 10- and 14-day old of both sexes were used. The parietal cortical neurons were acutely dissociated enzymatically. The extracellular fluid saturated with 0.3,0.6 and 0.9 mmol/L desflurane was added to the culture dish, then the effects of different concentrations of desflurane on Ik were investigated by using the whole-cell patch-clamp technique in acutely dissociated rat parietal cortical neurons. Results IK was inhibited by desflurane in a concentration-dependent manner ( P ＜0.01). The V1/2 of the activation and inactivation curves and the slop factor had no change after giving 0.6 mmol/L desflurane (P ＞ 0.05). Conclusion Desflurane inhibits delayed rectifier potassium channels of parietal cortical neurons of rats in a concentration-dependent manner, and has no effect on the activation and inactivation of delayed rectifier potassium channels, indicating that the change in the excitability of the channel is not involved in the mechanism of inhibitory effect of desflurane, and the other reasons may be involved in the mechanism.

Aim ToexploretheeffectsofnewdrugT-006 on improving learning and memory abilities in scopolamine-induced dementia mice and its possible mechanism.Methods 72maleKunmingmicewere randomly divided into six groups:normal control group,model group,donepezil treatment group,T -006 treatment group with different doses(1,3 and 10 mg·kg-1 ).All mice were treated by intragastric ad-ministration for 14 consecutive days. Learning and memory abilities were tested by a five-day Morris water maze trial from the 1 1 th day.the first 4 days of the five-day Morris water maze,the navigation test was performed,the last day of Morris water maze is the spatial probe test.During the navigation test, mice were intraperitoneally given 2 mg · kg-1 scopolamine 20 minutes before entering the water,while normal control group mice administrated with sterile saline in-stead.Mice were not given T-006 nor scopolamine in spatial probe test.After Morris water maze,all mice were sacrificed for hippocampus and cortex.The activi-ties of AchE and SOD and the levels of GSH and MDA in hippocampus and cortex were measured after tissue harvesting.Results Comparedwithmodelgroup,T-006 could obviously improve learning and memory abil-ities in scopolamine-induced mice, significantly in-crease the levels of SOD and GSH and decrease the levelsofMDAandAchE.Conclusion T-006can significantly improve cognitive abilities in scopolamine-induced dementia mice,and its relevant mechanism may be closely related to its antioxidative effect and the ability to decrease AchE level.

Objective:To investigate and evaluate the anti-tumor activity of hydroxychloroquine ( HCQ) in vitro. Methods:CCK-8 assay was applied to detect the inhibitory effect of HCQ at different concentrations(4. 78, 9. 55, 19. 10, 38. 20,76. 40 μg?ml-1 ) on A549 cells, HepG2 cells, HT-29 cells, K562 cells, Hela cells and B16 cells. Absorbance was detected by a microplate reader, and then the inhibitory rate of the tumor cells and the IC50 was calculated. Results:Compared with those of the negative control group, the inhibitory rates of HCQ at different concentrations against the six tumor cells were all increased significantly in a dose-dependent manner (P<0. 01). The inhibitory rates of the tumor cells were all above 60% when the concentration reached 38. 20μg?ml-1. The IC50 was 26. 70(A549), 27. 47(HepG2), 5. 72(HT-29), 14. 03(K562), 20. 21(Hela) and 13. 62(B16) μg?ml-1, respectively. Conclusion:HCQ shows anti-tumor activity on several cancer cells in vitro. There may be a good application prospect for HCQ in the treatment of colorectal cancer, liver cancer, cervical cancer, melanoma and leukemia.

Objective To investigate the cellular immune response to the recombinant DNA vaccine CRT120/HPV6bE7 in mice. Methods The recombinant encoding HPV6bE7, linked with CRT120, was constructed in pcDNA3.1 eukaryotic vector with the report gene GFP. The pcDNA3.1-GFP -HPV6bE7 was transfected into B16 cells by a lipofectamine kit. The C57BL/6 mice were vaccinated with recombinant DNA plamids. The T-cell phenotype in the peripheral blood lymphocytes of the immunized mice was measured by flow cytometry. The CTL activity of the lymphocytes from the spleens and lymph nodes, and the levels of IL-2 and IFN-? were analyzed. Results The constructed recombinant plasmid CRT120/HPV6bE7 was analyzed. Positive transfected cell clones were established and could stably express the target gene HPV6bE7. Compared with HPV6bE7, CRT120/HPV6bE7 plasmid enhanced to a greater extent CD8+ T-lymphocyte differentiation, the number of TCR?? T-lymphocytes and the levels of IL-2 and IFN-? (all P

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the surgical methods and clinical evaluation of complex tibial plateau fractures resulted from high-energy injuries.

METHODSFrom March 2006 to May 2009,48 cases with complex tibial plateau fractures were treated with open reduction and plate fixation, including 37 males and 11 females, with an average age of 37 years (ranged from 18 to 63 years). According to Schatzker classification, 16 cases were type IV, 20 cases type V and 12 cases type VI. All patients were examined by X-ray flim and CT scan. The function of knee joint were evaluated according to postoperative follow-up X-ray and Knee Merchant Rating.

RESULTSForty-eight patients were followed up with a mean time of 14 months. According to Knee Merchant Rating, 24 cases got excellent results, 16 cases good, 6 cases fair and 2 cases poor.

CONCLUSIONAppropriate operation time, anatomical reduction, suitable bone graft and reasonable rehabilitation exercises can maximally recovery the function of knee joint.

Adolescent ; Adult ; Bone Nails ; Bone Plates ; Bone Screws ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Internal Fixators ; Knee Joint ; surgery ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult