Objective: To elucidate the major reasons for unexpected death nude mice when they were used for establishing human metastatic cancer models. Methods: The fresh specimens of human renal cell carcinoma (RCC), prostate cancer (PC), and DU-145 cells were transplanted/injected into nude mice ecotopically and orthotopically. Tumorigenesis and pathological changes (including the symptoms, pathological sections, survival time, etc.) of mouse liver were investigated subsequently. Results: The tumorigenesis and metastasis rates were respectively 21.7% (35/161) and 1.2% (2/161) after implantation of RCC sample, and were respectively 100% (20/20) and 25% (5/20) after implantation of DU-145 cell line, while there was no tumorigenesis or metastasis after implantation of PC specimens. Liver pathological changes were found in 58.4% (94/161) of mice implanted with RCC samples and in 43.4% (46/106) of mice implanted with PC samples. No pathological lesion was found in mice implanted with DU-145 cells. The death peak of mice with pathological changes after implanting RCC and PC samples was consistent with that of the total mice used in this study, all occurring in the winter and spring of the year. Conclusion: The pathological changes of liver appear to be the major reason of unexpected death of the nude mice when they were used for establishing human metastatic cancer models. A specified pathogen-free environment is very important for establishment of the models.

Objective: To establish orthotopic mouse models of human renal cell carcinoma and to separate metastatic or non-metastatic renal cell carcinoma (RCC) from the same source. Methods: Surgical specimens/cell suspensions were transplanted into the various tissues of BALB/c nude mice (subcutis, cellular orthotopic injection into renal capsule, perinephrium, surgical orthotopic implantation into renal capsule). Tumorigenicity and metastasis were subsequently evaluated. Immunohistochemical analysis was employed to determine the expression of VEGF, bFGF, P16, Bcl-2 and C-met in the metastatic and non-metastatic RCC xenograft. Results: The incidences of tumorigenicity and metastasis of orthotopic model were the highest, being at 73.3% (11/15) and 20% (3/15), respectively. Compared with the non-metastastic RCC, VEGF expression was upregulated in the metastatic RCC (P<0.05), while the expression of C-met was downregulated significantly (P<0.05); the expression of bFGF, Bcl-2 and P16 was also downregulated but without significance. Conclusion: Mouse orthotopic transplantation is the most effective way for the growth and natural metastasis of human RCC. Primary and metastatic RCC from the same patient has been successfully harvested. Expression of VEGF in the metastatic tumor tissues is significantly higher than that in the non-metastatic RCC.

Objective: To isolate the invasive and non-invasive cells from primary human renal cell carcinoma (RCC) in vitro. Methods: Fresh RCC surgical specimens from 32 primary RCC patients were primarily cultured following enzyme digestion or mechanical minimization in vitro. In vitro invasion assay using the Transwell cultures coating Matrigel was performed for separation and recovery of invasive and non-invasive cells from the primary culture of 3 RCC patients. The concentration of Matrigel, recovery time and trypsinization were subsequently optimized. Results: The successful rate of primary culture was 90.6% (29/32). Recovery of invasive cells was performed ideally when matrigel (diluted into 1.0 mg/ml and 20 μl) was coated onto the filter of the well; cell suspension was at a concentration of 5 × 105/ml and invasive cells were recovered on the 5th day of culture. The growth of non-invasive cells was scattered, while that of the invasive cells was focal. The doubling time of invasive cells was 36.1 h and that of non-invasive was 50.6 h. Conclusion: The in vitro invasion assay using the Transwell is able to separate and recover the highly invasive primary RCC cells. The primary cells represent intact subpopulation composition, but it can hardly get through the life span of human primary tumor cells.

BACKGROUND:Bone marrow is the main source of mesenchymal stem cells(MSCs)at present,but its application has been limited,because of some reasons such as inconvenience of isolation,and the quantity of cells decreases with human increased age.Umbilical cord as a new source of MSCs has been widespread concerned recently.OBJECTIVE:To explore the approach of isolating and culturing MSCs from Wharton's jelly of human umbilical cord,and the methods of identifying the surface antigens and the differentiation potential.METHODS:MSCs were isolated and amplified via tissue-cultivation,and cultured by FasGrow medium.Morphology of MSCs from Wharton's jelly of human umbilical cord was observed under the optical microscope.Its immunophenotypes were detected using immunohistochemistry.The differentiation of MSCs into the osteoblasts was determined utilizing Gomori calcium-cobalt alkaline phosphatase staining,von Kossa calcium node staining,and tetracyclinefluorescence labeling.The differentiation of MSCs into the adipocytes was detected using oil red O staining.RESULTS AND CONCLUSION:MSCs were easily obtained from Wharton's jelly of human umbilical cord via the proposed approach.The primary cells grew up to 70%-80% confluence after 12-16 days of culture,and meanwhile the undifferentiated state was maintained and proliferation was stabilized after passage.The cell cycle of double increase was about 2 days,and proliferation in vitro reached twenty generation above.Surface antigen analysis showed that CD44,CD105,CD133,MHC-I were positively expressed,while CD34,CD45 were negatively expressed.Experiments of differentiation in vitro indicated that the obtained cells were capable of differentiating into fat,osteoblast and nerve-like cells.

BACKGROUND:Platelet-rich plasma (PRP) containing high levels of platelet-derived growth factor for knee osteoarthritis has achieved good clinical results;however, the effects of RPR on the repair of damaged articular cartilage are stil controversial.
OBJECTIVE:To study the effects of RPR on the repair of damaged articular cartilage in a rabbit model of knee osteoarthritis.
METHODS:The model of osteoarthritis in the rabbit right knee was established by Hulth’s method. Autologous PRP (0.5 mL) (PRP group), sodium hyaluronate (0.5 mL) (sodium hyaluronate group), and normal saline (model group) were injected into the right knee joint cavity, respectively. The morphology of articular surface and nitric oxide contents in knee joint fluid were observed and determined at 8 weeks after surgery.
RESULTS AND CONCLUSION:The morphology of articular cartilage in the PRP group was better than that in the other three groups. Mankin scores of articular cartilage and nitric oxide contents of knee joint fluid in the PRP group were significantly decreased compared with the model and sodium hyaluronate groups that in (P<0.05), while increased compared with the sham-operation group (P<0.05). Our results suggest that repair effects of PRP on the damaged articular cartilage are superior to sodium hyaluronate treatment.

In recent years,the course of pathogenic biology of Nanhua University has made great achievements because the discipline characteristics has been stressed.Therefore,the reputation of Nanhua University is improved with the elevation of this discipline.The development patterns of Nanhua University provide some experience for the building and development of general university.

Objective To investigate the treatment mechanism and effect of platelet-rich plasma(PRP) in treating rabbit knee osteoarthosis(KOA).Methods Thirty-two New Zealand adult clean class rabbits were randomly divided into the blank control group(A),model control group(B),PRP treatment group(C) and sodium hyaluronate treatment group(D).The group A conducted the simulated model construction,while the group B,C and D established the KOA rabbit model by using the Hulth method.After establishing the animal model,the group C was given PRP 0.5mLl by knee articular cavity injection,once every 3 weeks,twice in total;the group D was given sodium hyaluronate by knee articular cavity injection,once per week for 5 continuous weeks.The group A and B were injected with equal amount of normal saline at the same time point in the group C.The histological structure of articular cartilage,cell number,integrity of tidal line,and toluidine blue staining were observed by common optical microscope.The differences of Mankin′s scores were compared among various groups.The proper amount of knee joint fluid was collected for measuring the arachidonic acid expression in each group.Results The structure and morphology of articular cartilage in the group A were normal without obvious damage,while which in the group B,C and D had different degrees of damage,especially the cartilage structure in the group B had greater changes.Although the group B and C had the morphological and structure change of articular cartilage,but which was close to the cartilage structure in the group A.The Mankin′s score in the group A was lowest,while which in the group B was highest,which in the group C and D was significantly decreased after intervention treatment;the difference between the group B with the group C and D had statistical significance(P<0.05).The difference between the group C and D had statistical significance(P<0.05).The arachidonic acid level in the group A was lowest and which in the group B was highest,which in the group C and group D ranged between the group A and B,moreover the group C was lower than the group D (P<0.05).Conclusion PRP has obvious therapeutic and alleviated effect in treating KOA.

In the development of researched experiment teaching,we constructed favorable atmosphere by establishing innovation idea and construction pattern,emphasizing the integration of teaching content,scientific research and society practice,the innovation of establishing experiment staff scientifically and systemically.

OBJECTIVETo explore the correlation between hand and face-mouth, so as to provide nerve reflex basis for the theory "Hegu (LI 4) regulates face and mouth".

METHODSSeven hundred and sixty-three participants who met the inclusive criteria were divided into different age groups. The skin around participants' thenar eminence was gently scraped to be observed whether there was an involuntary movement around the face or mouth, which was palmomental reflex. The results of palmomental reflex were recorded.

RESULTSThe total occurrence rate of palmomental reflex was 46.26%. For those who were 0 to 1 years old, the palmomental reflex was all positive; for those who were 21 to 36 years old, the positive rate was 20.45%, which was the lowest; for those who were 65 to 85 years old, more than half of them were positive. The majority of those who were 0 to 2 years old were bilateral positive palmomental reflex, while the majority of those who were 65 to 85 years old were unilateral positive palmomental reflex.

CONCLUSIONThere is a certain connection between hand and face-mouth. The occurrence rate of palmomental reflex changes from high to low over age increasing, and then changes from low to high with the aging, presenting a "high-low-high" U-shaped curve, which is possible related to the growth and recession of nervous system.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Face ; physiopathology ; Female ; Hand ; physiopathology ; Humans ; Infant ; Male ; Middle Aged ; Mouth ; physiopathology ; Reflex ; Young Adult

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection.Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins ( Core, E1 and E2) de-rived from a Chinese HCV patient (genotype 1b, Hebei strain).Enzyme linked immunospot assay (ELIS-POT) and intracellular cytokine staining ( ICS) were used to analyze the elicited antigen-specific immune re-sponses and the efficacy of cross-protection.Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vac-cines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT as-say.ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+and IFN-γ-produ-cing CD8+T cells against E1 protein and high levels of IFN-γ-producing CD4+and CD8+T cells against E2 protein in comparison with immunization with DNA vaccines.Moreover, the prime-boost vaccination was ca-pable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterolo-gous HCV (JFH1, 2a) vaccinia virus.Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV in-fection.