Objective To study the expression of p16 and Rb gene in primary gallbladder carcinoma (PGC). MethodsFiftysix cases of PGC and 26 of gallbladder benign lesions were observed by immunohistochemistry (SP). ResultsThe positive rates of p16 and Rb protein in PGC were 46.3% and 71.4% respectively. No association of Rb expression with histological grade, histological type, invasion, metastasis and prognosis was found. There were close correlation between p16 expression and invasion, metastasis as well as prognosis. The inverse correlation of p16 and Rb gene expression in PGC was confirmed. Conclusion The loss of p16 or Rb gene protein is an important molecular event in PGC,and closely related to the block in the negative regulatory pathway of all cell cycle. The detection of p16 protein may be useful in evaluating biological characteristics and prognosis in PGC.
OBJECTIVE: To investigate the effects of three-dimentional culture in bioderived material modified by Pluronic F-127 on the growth and function of rabbit periosteal osteoblast in vitro. METHODS: Bio-derived materials were from fresh pig ribs and were modified by Pluronic F-127. Then rabbit periosteal osteoblasts were cultured in bio-derived materials(group A), in the modified bio-derived materials(group B) and on the plastic surfaces as a control (group C), respectively. During a 7-day period, the status of growth, cell viability and alkaline phosphatase(ALP) activity were measured. RESULTS: Osteoblasts attached, elongated and grew well on the modified bio-derived materials. There were no significant difference in osteogenesis and ALP activity between group A and group B(P gt; 0.05). The osteogenesis and ALP activity in groups A, B were less than those in group C (P lt; 0.01). CONCLUSION: Pluronic F-127 can be used for a carrier for bioactive factors to modify bio-derived material.
Objective To detect the expression of thromhospondin-1 （TSP-1） in gastric cancer and metastaticlymph node tissues， and to study its relationship of TSP-1 to clinicopathologic parameters or tumor angiogenesis. Methods The TSP-1 and vascular endothelial growth factor （VEGF） expressions and microvessel density （MVD） were evaluated by immunohistochemistry in 72 specimens obtained by gastric resection from patients with gastric cancer， including corres-ponding adjacent normal gastric mucosa tissues （distant from cancer ≥5 cm） and lymph nodes surrounding cancer. A semiquantitative scoring system was used for evaluating the staining. The relationship of TSP-1 to VEGF expression， MVD， or clinicopathologic parameters was analyzed. Results ① TSP-1 positive expression rate was 45.8% （33/72） in the primary gastric cancer tissues， 90.3% （65/72） in the corresponding adjacent normal gastric mucosa tissues， and 50.8% （30/59） in the metastatic lymph nodes tissues. The expressions of TSP-1 in the primary gastric cancer tissues and metastatic lymph nodes tissues were significantly lower than those in the adjacent normal gastric mucosa tissues （χ2=32.710，P=0.000；χ2=25.298， P=0.000）. The expression of TSP-1 had no statistical significance in the primary gastric cancer tissues as compared with in the metastatic lymph nodes tissues （χ2=0.327， P=0.568）. ② The expression of TSP-1 in the metastatic lymph nodes tissues was significantly lower than that in the non-metastatic lymph nodes tissues （Z=-2.573， P=0.010）. ③The expression of TSP-1 in the primary gastric cancer tissues and metastatic lymph nodes tissues suggested a negative correlation with VEGF （rs=-0.309， P=0.008；rs=-0.269， P=0.040） and MVD （rs=-0.348， P=0.003；rs=-0.272， P=0.037）. Conclusions TSP-1 expression is down-regulated and has a negative correlation with VEGF and MVD in the primary gastric cancer and the metastatic lymph nodes tissues. According to the present results， it seems likely that TSP-1 is a tumor angiogenesis inhibitor.
Objective To investigate the heterotopic odontogenesis ability ofDelta1 gene transfected human dental pulp stem cell (DPSC) and nanohydroxyapatite/collagen (nHAC) composite scaffold. Methods The cultured human DPSC was transfected with Delta1-enhanced green fluorescent protein recombinant retrovirus supernatant，and was selected by puromycin to obtain the positive cell clone. The experimental group contained the Delta1 transfected DPSC; however, the control group did not contain the Delta1 transfected DPSC but contained DPSC transfected with vectors only. The cells were seeded into the nHAC carriers and were cultured in the odonto-inductive medium. The growth of the transduced cells in the carriers was observed by the fluorescent phase contrast microscope and the scanning electron microscope (SEM). The cell-carrier composites were subcutaneously transplanted into the Delta1 transfected 8 nude mice (female, 8 weeks old). Eight weeks after operation,the composites were taken out and tested with the histological and the immunohistological methods.Results Green fluorescence was observed inthe cells in the experimental group, which were grown in the carriers by the fluorescent phase contrast microscope. Observed by SEM, great amounts of transduced DPSC were observed along the scaffold materials, even filling the porous structures of nHAC and secreting a lot of extracellular matrix. However, in the control group, much fewer cells were found in the carriers. All the 4 Delta1 transduced DPSC-nHAC composites produced dentin-like structures that lined the surfaces of some nHAC porous structures. The odontoblast-like cells extended the cytoplasmic processes into the dentinal matrix, which was interfaced with a pulp-like interstitial tissue infiltrated with the blood vessels. Dentin sialophosphoprotein was expressed in the odontoblast-like cells when immunohisochemistry was performed. The morphology of the control composite was a typical one of the fibrous connective tissue,and only a little dentin-like structure was found in 2 of the 8 control transplants. Conclution DPSC can be used as the recipient cell of the Delta1 gene for expression and secretion of the Delta1 protein. The composites of the transfected cells and nHAC can induce heterotopic odontogenesis, which indicates that Delta1 is a novel candidate for the gene enhanced dentinpulp composite engineering.
Since the beginning of the coronavirus disease 2019 (COVID-19) pandemic at the end of December 2019, more than 85% of the population in China has been infected. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mainly affects the respiratory system, especially the lungs. The mortality rate of patients with severe infection is high. A percentage of 6% to 10% of patients will eventually develop into COVID-related acute respiratory distress syndrome (CARDS), which requires mechanical ventilation and extracorporeal membrane oxygenation (ECMO) support. Some patients who survive acute lung injury will subsequently develop post COVID-19 pulmonary fibrosis (PCPF). Both fully treated CARDS and severe PCPF are suitable candidates for lung transplantation. Due to the special course, evaluation strategies are different from those used in patients with common end-stage lung disease. After lung transplantation in COVID-19 patients, special treatment is required, including standardized nucleic acid testing for the novel coronavirus, adjustment strategy of immunosuppressive drugs, and rational use of antiviral drugs, which is a big challenge for the postoperative management of lung transplantation. This consensus was evidence-based written and was reached by experts after multiple rounds of discussions, providing reference for assessment and postoperative management of patients with interstitial pneumonia after COVID-19 infection.
ObjectiveTo study immunodepression effect of bone marrow-derived mesenchymal stem cell (BMSC) on acute asthmatic airway inflammation by galectin-1 (gal-1) in vivo.MethodsEighty-five female BALB/c mice were equally randomized into normal control group, asthmatic group, BMSC treatment group, gal-1 treatment group and BMSC and gal-1 inhibitor group. Ovalbumin (OVA) was used to establish acute asthmatic model. Total cell number and differential cell analysis in each group in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, hematoxylin-eosin and periodic-acid Schiff staining was used to compare airway inflammation among five groups. Measurement of cytokines, including interleukin (IL) -4, IL-5 and gal-1 in BALF and OVA specific IgE (OVA-IgE) in serum were evaluated by enzyme linked immunosorbent assay. Moreover, dendritic cell (DC) in lung tissue was sorted by immunomagnetic beads and its MAPK signal pathway was analyzed by western blotting among five groups.ResultsAccumulation of inflammation cells, particularly eosinophils around airway and in BALF was evident in asthmatic mouse model, meanwhile hyperplasia of Goblet cell was also obvious in asthmatic group. BMSC engraftment or gal-1 infusion significantly reduced airway inflammation and hyperplasia of Goblet cell and the number of inflammation cells in BALF, especially eosinophils attenuated dramatically. However, there was no effect on airway inflammation and hyperplasia of Goblet Cell by simultaneous infusion BMSC engraftment and gal-1 inhibitor. Compared to normal control group, the level of IL-4, IL-5 in BALF and OVA-IgE in serum was increased remarkably in asthmatic group, but the level of gal-1 reduced obviously. Moreover, infusion of BMSC or gal-1 could mitigate the level of IL-4, IL-5 in BALF and OVA-IgE in serum and increase the level of gal-1 in asthmatic mouse. However, infusion with both BMSC and gal-1 inhibitor exerted no effect on cytokine and OVA-IgE in asthmatic mouse. DC was sorted by immunomagnetic beads and western blotting was used to detect the expression of MAPK signal pathway among five groups. The expression of ERK phosphorylation in asthmatic group was much lower than that in normal control group. On the contrary, the expression of p38 phosphorylation was much higher than that in normal control group. BMSC engraftment or gal-1 infusion significantly activated the ERK pathway and inhibited the p38 MARP pathway on asthmatic mouse DC. Nevertheless, the expression of ERK phosphorylation and p38 phosphorylation for group with BMSC and gal-1 inhibitor infusion was between the level of asthmatic group and normal control group.ConclusionsBMSC infusion alleviates airway inflammation in asthmatic mouse, especially weakens eosinophils infiltration, and the underlying mechanism might be protective effect of gal-1 secreted by BMSC which plays a role in lung tissue DC and regulates the DC expression of MAPK signal pathway.
Objective To analyze the clinical features and treatment of severe H1N1 influenza.Methods The clinical data of 34 patients with severe H1N1 influenza admitted to intensive care unit from October to December 2009 were reviewed. Results The patients aged 3 months to 60 years with an average of ( 13. 9 ±4. 5) years, of which 24 patients were younger than 7 years old. Fever( 30 cases) , cough( 32 cases) , progressive shortness of breath( 19 cases) were the main symptoms. White blood cell count was normal in 21 cases, increased in 6 cases, and decreased in 7 cases. Lymphocyte count was normal in 16 cases, increased in 12 cases, and decreased in6 cases. Chest X-ray films showed bilateral or unilateral patchy pulmonary fuzzy shadows in28 cases. Chest CT showed diffuse interstitial lesion in1 case, pleural effusion in 2 cases, and bronchiectasis in 1 case. The hepatic and myocardial enzymogramparameters were all abnormal.30 cases were treated by oseltamivir and ribavirin, 4 cases by methyllprednisolone, and 6 cases by gamma globulin. 8 cases underwent routine intubation and mechanical ventilation, and 5 cases received non-invasive mechanical ventilation. All 34 patients were cured. Conclusions Lung, heart, and liver are the major target organs in severe H1N1 influenza. Mechanical ventilatory support is an important treatment for severe H1N1influenza.
Objective To observe the histopathologic features and expression patterns of tumor necrosis factor-alpha; （TNF-alpha;）, interleukin-1beta;（IL-1beta;） and Escherichia coli lipopolysaccharide (LPS) in the rat vitreous with LPS inducedendophthalmitis. Methods Wistar rats were randomly divided into saline control group (SC,136 rats）,endophthalmitis group （EO, 168 rats）and blank control group （BC,12 rats）.EO group received an intravitreal injection of 5 mu;l LPS; SC group received 5 mu;l sterile saline and no intervention for BC group.Six,12,24,48, and 72 hours,5 and 7 days after injection, intraocular inflammation were observed and the eyes and vitreous were collected for histopathological examination and measurement of TNF-alpha;, IL-1beta; and LPS expression. Results Severe inflammatory responses in the eyes were observed in EO group between six and 72 hours after LPS injection，ocular inflammation subsided seven days after LPS injection. In the vitreous, a peak neutrophil count was observed at 24 hours （1224.64plusmn;132.2） cells/eye that rapidly declined at 72 hours （342.25plusmn;47.7） cells/eye. The levels of TNF-alpha; and IL-1beta; in EO group were peaked at 24 hours with （996.18plusmn;89.45） and（5556plusmn;1440）pg/ L, respectively；Persisted at 48 hours and began to decline rapidly thereafter. Seven days after LPS injection, levels of TNF-alpha; and IL-1beta; returned to baseline with （22.16plusmn;5.84）and （73.7plusmn;18.7） pg/L, respectively. LPS concentration in EO group decrease rapidly at 72 hours with （11.03plusmn;3.41） ng and disappear on days 7 with (0.22plusmn;0.08) ng after LPS injection.Conclusions Massive neutrophils infiltration, high levels expression of TNF-alpha; and IL-1beta; and spontaneous elimination of bacterial elements in vitreous cavity were major pathologic characteristics in this experimental model. The expression patterns of TNF-alpha;,IL-1beta; were in accord with LPS clearance process.
Objective To transfect bone marrow mesenchymal stem cells (BMSCs) of rats by recombinant adenovirus Ad-human matrix metalloproteinase 1 (hMMP-1) in vitro so as to lay the experimental foundation for the treatment of liver fibrosis with a combination of BMSCs and hMMP-1 gene transplantation. Methods BMSCs were isolated from bone marrow of 2-3 weeks old Sprague Dawley rats by whole bone marrow adherence method and identified, then transfected by recombinant adenovirus Ad-hMMP-1 carrying enhanced green fluorescent protein (EGFP) marker in vitro. The green fluorescent expression was observed by fluorescence microscope and the transfection efficiency was detected by flow cytometry to determine the optimum multiplicity of infection (MOI). BMSCs at passage 3 were divided into 3 groups: untransfected BMSCs group (group A), Ad-EGFP transfected BMSCs group (group B), and Ad-hMMP-1-EGFP transfected BMSCs group (group C); the gene and intracellular protein of hMMP-1 were detected by RT-PCR and Western blot; the ELISA assay was used to detect the supernatant protein expression, and the hMMP-1 activity was measured by fluorescent quantification kit. Results The green fluorescent was observed in BMSCs transfected by recombinant adenovirus at 24 hours after transfection; the fluorescence intensity was highest at 72 hours; and the optimum MOI was 200. The cells of 3 groups entered the logarithmic growth phase on the 3rd day and reached plateau phase on the 6th day by MTT assay; no significant difference was found in the cell proliferation rate among 3 groups (P gt; 0.05). RT-PCR, Western blot, and ELISA assay showed high expressions of the hMMP-1 gene and protein in group C, but no expression in groups A and B. The hMMP-1 activity was 1.24 nmol/(mg · min) in group C, but hMMP-1 activity was not detectable in groups A and B. Conclusion The exogenous hMMP-1 gene is successfully transfected into BMSCs of rats via recombinant adenovirus and can highly express, which lays the experimental foundation for the treatment of liver fibrosis with a combination of BMSCs and hMMP-1 gene transplantation.
Objective To summarize the effect of lenvatinib + transarterial chemoembolization (TACE) + programmed cell death protein-1 (PD-1) antibody in the treatment of hepatocellular carcinoma with main portal vein tumor thrombus and cavernous transformation. Methods In this study, we reported the clinical data of four patients with hepatocellular carcinoma with main portal vein tumor thrombus and cavernous transformation who received conversion therapy with lenvatinib combined with TACE and PD-1 antibody in West China Hospital. Results Among the four patients, two patients achieved complete response and two achieved partial response; tumor markers were significantly decreased after combination treatment. However, all four patients failed to undergo hepatectomy. ConclusionsLenvatinib + TACE + PD-1 antibody is effective for hepatocellular carcinoma with main portal vein tumor thrombus and cavernous transformation. However, there are still many problems worthy of further discussion.