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OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.
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Animals , Humans , Mice , Carcinoma, Hepatocellular/pathology , Cyclophosphamide/therapeutic use , Hemolysin Proteins , Heterografts , Interleukin-2 , Liver Neoplasms/pathology , Panax notoginseng , Saponins/therapeutic use , Tumor Necrosis Factor-alphaABSTRACT
Objective@#To investigate the expression of syndecan-1 and syndecan-2 and their clinicopathological significance in patients with gallbladder squamous cell (SC)/adenosquamous carcinoma (ASC) and adenocarcinoma (AC).@*Methods@#A total of 126 patients with SC/ASC (n=46) and AC (n=80) were included in this study. The expression levels of syndecan-1 and syndecan-2 were detected by Envison™ immunohistochemistry assay. The clinical and prognostic significance of syndecan-1 and syndecan-2 were analyzed.@*Results@#In the 46 SC/ASC samples, syndecan-1 and syndecan-2 were positively expressed in 29 (63.0%) and 28 (60.9%) tumor tissues, respectively. (Positive expression was defined based on the staining in the component of squamous cell carcinoma. That is to say, the tissue which adenocarcinoma part was positively stained, but squamous cell carcinoma part was negatively stained is also regarded as negative.) In the 80 AC samples, 47 (58.8%) cases showed syndecan-1 positive expression, and 51 (63.8%) showed syndecan-2 positive expression. There was no significant difference in the positive rates of syndecan-1 and syndecan-2 between SC/ASC and AC groups (P>0.05 for all). The levels of syndecan-1 and syndecan-2 were associated with tumor size, TNM staging, lymph node metastasis, invasion of adjacent tissue, and surgical procedures in SC/ASC patients (P<0.05 for all). However, their expression was associated with tumor differentiation, tumor size, TNM staging, lymph node metastasis, invasion of adjacent tissue, and surgical procedures in AC patients (P<0.05 for all). The Kaplan-Meier survival analysis of SC/ASC and AC patients revealed that the average survival time for patients with positive syndecan-1 and syndecan-2 expression was significantly shorter than that of those with negative expression (P<0.01 for all). Cox multivariate analysis indicated that syndecan-1 and syndecan-2 expression were independent unfavorable prognostic factors for SC/ASC and AC patients (P<0.05 for all).@*Conclusion@#The syndecan-1 and syndecan-2 expression are associated with the tumor progression and poor prognosis in patients with gallbladder SC/ASC and AC.
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Objective To investigate the value of delayed 18F-FDG PET/CT with oral intake small dosage diuretics for diagnosing urogenital cancers.Methods Patients with suspected urogenital system cancers were divided into routine dosage diuretic group (n=12) and small dosage diuretics group (n=35).All patients underwent whole-body PET/CT followed by delayed scanning after oral 40 mg or 20 mg Furosemide respectively.The urine maximum standard uptake value (SUVmax) and T/U (the ratio of urine and lesion SUVmax) before and after diuresis were compared respectively.Diagnostic efficacy for malignant urogenital system cancers of small dosage group was calculated.Results The urine SUVmax and T/U were statistically different between routine whole body and delayed scans in both groups (P＜0.05).SUVmax and T/U of routine and delayed scans had no statistical differences between the two groups (P＞0.05).In small dosage group,the sensitivity,specificity,positive predictive value,negative predictive value and accuracy of delayed imaging and routine imaging was 96.77％ (30/31)and 61.29％ (19/31),75.00％ (3/4) and 50.00％ (2/4),96.77％ (30/31) and 90.48％ (19/21),75.00％ (3/4)and 14.29％ (2/14),94.29％ (33/35) and 60.00％ (21/35),respectively.The sensitivity and accuracy were statistically different between routine and delayed imaging (P＜0.001).Conclusion Delayed PET/CT imaging with oral small dosage Furosemide has the same efficacy as PET/CT using routine dosage diuretics,which is useful for diagnosing urogenital cancers.
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Objective@#To study the effects of seawater immersion on the inflammatory response and oxygen free radical injury of rats with superficial-thickness scald at early stage.@*Methods@#Seventy Wistar rats were divided into healthy control group (HC, n=7), pure scald group (PS, n=21), scald+ fresh water immersion group (SF, n=21), and scald+ seawater immersion group (SS, n=21) according to the random number table. Rats in group HC did not receive any treatment, while 5% total body surface area superficial partial-thickness scald was made on the back of rats in the latter three groups. Rats in group PS lived freely immediately post burn, while wounds on the back of rats in groups SF and SS were immersed into fresh water and seawater, respectively. Serum and full-thickness skin tissue in the center of wounds on the back of 7 rats in groups PS, SF, and SS at post immersion (injury) hour (PIH) 2, 4, and 6 were collected, respectively, while serum and full-thickness skin tissue at the same position of the 7 rats in group HC were collected at PIH 6 of rats in other groups. Morphology of skin tissue was observed with HE staining; tumor necrosis factor-alpha (TNF-α) content in serum and skin tissue was determined by enzyme-linked immunosorbent assay; superoxide dismutase (SOD) content in serum and skin tissue was determined by hydroxylamine method; malondialdehyde content in serum and skin tissue was determined by thiobarbituric acid method. Data were processed with analysis of variance of factorial design, one-way analysis of variance, Welch test, LSD test, and Tamhane test.@*Results@#(1) Epidermal cells of skin tissue of rats in group HC arranged in order and continuously, and the dermis tissue and accessory structures were clear and complete. The skin layer and epidermis of wounds of rats in group PS had no significant change, but the edema of epidermis and dermis and infiltration of inflammatory cells enhanced over time at PIH 2, 4, and 6. The horny layer of epidermis of wounds of rats in group SF reduced, and the edema of epidermis and dermis and infiltration of inflammatory cells enhanced over time at PIH 2, 4, and 6; some epidermal cells disintegrated at PIH 6. The horny layer of epidermis of wounds of rats in group SS significantly reduced, along with the increase in disintegration of epidermal cells, the significant enhancement of edema of epidermis and dermis, and infiltration of a large number of inflammatory cells over time at PIH 2, 4, and 6. (2) Compared with (247±27) pg/mL in group HC, the serum content of TNF-α of rats in group PS significantly increased at PIH 2 and 4 [respectively (675±122) and (367±54) pg/mL, P<0.05 or P<0.01] but significantly decreased at PIH 6 [(147±27) pg/mL, P<0.01]; the serum content of TNF-α of rats in group SF significantly decreased at PIH 6 [(90±24) pg/mL, P<0.01]; the serum content of TNF-α of rats in group SS significantly increased at PIH 2, 4, and 6 [respectively (1 646±58), (2 086±114), and (2 951±58) pg/mL, with P values below 0.01]. Compared with (364±123) U/mL in group HC, the serum content of SOD of rats in group PS significantly increased at PIH 2 and 4 [respectively (489±13) and (447±14) U/mL, with P values below 0.05]; the serum content of SOD of rats in group SF significantly decreased at PIH 6 [(282±13) U/mL, P<0.05]; the serum content of SOD of rats in group SS significantly increased at PIH 2 [(461±23) U/mL, P<0.05] but significantly decreased at PIH 4 and 6 [respectively (226±8) and (205±10) U/mL, with P values below 0.01]. Compared with that in group HC, the serum content of malondialdehyde of rats in groups PS, SF, and SS significantly increased at PIH 2, 4, and 6 (with P values below 0.01). (3) Compared with that in group HC, the TNF-α content in wound tissue of rats in groups PS and SS significantly increased at PIH 2, 4, and 6 (P<0.05 or P<0.01), and the TNF-α content in wound tissue of rats in group SF significantly increased at PIH 2 and 4 (with P values below 0.01). Compared with that in group HC, the SOD content in wound tissue of rats in groups PS and SF significantly increased at PIH 2, 4, and 6 (P<0.05 or P<0.01), and the SOD content in wound tissue of rats in group SS significantly increased at PIH 2 and 4 (with P values below 0.01). Compared with that in group HC, the malondialdehyde content in wound tissue of rats in groups PS, SF, and SS significantly increased at PIH 2, 4, and 6 (with P values below 0.01).@*Conclusions@#Seawater immersion can enhance the inflammatory response and oxygen free radical injury of wounds and the whole body of rats with superficial partial-thickness scald at early stage.
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OBJECTIVE:To investigate the compatible stability of cefpirome sulfate with ornidazole. METHODS:At room temperature [(20±1)℃],the appearances(color,clarity,sedimentation and gas)and pH changes of the mixtures by Cefpirome sulfate for injection with Ornidazole for injection in 0.9% Sodium chloride injection and 5% Glucose injection after 0,1,2,4,6 h were observed. RP-HPLC was adopted to determine its content changes. RESULTS:No significant change was noted for the mix-ture in appearance within 6 h,pH value ranged in 3.359-3.588;compared with the beginning(0 h),the contents of cefpirome sul-fate and ornidazole ranged in 100.2%-100.3%,99.9%-100.4% in 0.9% Sodium chloride injection at each time point,as well as 99.7%-99.9%,99.4%-99.6%in 5%Glucose injection. CONCLUSIONS:At room temperature,cefpirome sulfate mixed with orni-dazole show stable appearance,pH value and content in 0.9%Sodium chloride injection and 5%Glucose injection within 6 h.
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<p><b>OBJECTIVE</b>To screen the genes related with leukocyte responses in mice early after burn injury by bioinformatic analysis of the gene expression profiling data.</p><p><b>METHODS</b>The gene expression profiles were obtained from GEO (GSE7404, Mouse musculus, 25% TBSA, full-thickness) database. T test, fold changes and GO functional enrichment analysis were used to identify the differentially expressed genes (DEGs) related to leukocyte responses to burns; the interacting genes were transferred to STRING to construct the protein-protein interaction (PPI) network. Biological annotation of the sub-networks was executed using the software Cytoscape. Real-time PCR and Western blotting were used to verify the DEGs in mice.</p><p><b>RESULTS</b>In mice at 1 day post-burn, a total of 658 genes were up-regulated and 1167 were down-regulated. PPI network and module analysis suggested that some of the genes (Stat1, Cdk1, Cd19, Lck and Jun) may play critical roles in the PPI network post-burn. Real-time PCR and Western blotting results in mice were consistent with those of bioinformatic analysis of Stat1, Cdk1 and Jun.</p><p><b>CONCLUSION</b>Stat1, Cdk1 and Jun might be critical players in the development of leukocyte response in mice early after burn injury. Our finding provides new insights into the pathogenesis of leukocyte response to burn injury and identifies several biomarkers as potential targets for burn treatment.</p>
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[Objective]To radiolabel the PSMA aptamer A10-3.2 with 99mTc , and explore its biological characteristics in vivo and in vitro.[Methods]Using Succinimidyl 6-hydrazinonicotinate hydrochloride (SHNH) as the bifunctional chelating agent to label aptamer A10-3.2 with 99mTc, then tested for the stability in vitro, the specific uptake by prostate cancer LNCaP cells (PSMA+) , the characteristics of SPECT/CT imaging and biodistribution in LNCaP tumor-bearing NOD/SCID mice.[Results]The labeling rate and radiochemical purity of the products (99mTc-SHNH-A10-3.2) are(71.31 ± 6.78)% and 97.03%,respectively. 99mTc-SHNH-A10-3.2 had obvious target specificity for PSMA positive prostate cancer LNCaP cells, its uptake rate was significantly higher than PSMA nega?tive PC-3 cells (P<0.01). And in tumor-bearing mice, the tumor has a certain uptake and a high ratio of the tumor tissue to the mus?cle.[Conclusion]This study successfully constructed 99mTc-labeled PSMA-targeted aptamer A10-3.2, which has a good stability and targeting in vivo and in vitro, has a high tumor tissue/muscle ratio in tumor-bearing mice, which show that it may be a potential target?ed molecular imaging agent for prostate cancer.
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Objective To investigate the effects of mechanical strain on Ca-calmodulin dependent kinase (CaMK)-cAMP response element binding protein (CREB) signal pathway and proliferation of osteoblasts.Methods Using a four-point bending device, MC3T3-E1 cells were exposed to mechanical tensile strains of 2500 µs and 5000 µs at 0.5 Hz respectively. The intracellular free Ca([Ca]i) concentration and calmodulin activity were assayed by fluorospectrophotometry, CaMK II β, CREB, and phosphorylated (activated) CREB (p-CREB) were assessed by Western blot, and cells proliferation was assayed with MTT. Pretreatment with verapamil was carried out to block Cachannel, and inhibitor U73122 was used to inhibit phospholipase C (PLC).Results Mechanical strains of 2500 µs and 5000 µs for 1 to 10 minutes both increased [Ca]i level of the cells. The 2500 µs strain, a periodicity of 1 h/d for 3 days, activated calmodulin, elevated protein levels of CaMK II β and p-CREB, and promoted cells proliferation, which were attenuated by pretreatment of verapamil or U73122. The effects of 5000 µs strain on calmodulin, CaMK II β, p-CREB and proliferation were contrary to 2500 µs strain.Conclusion The mechanical strain regulates osteoblasts proliferation through Ca-CaMK-CREB signal pathway via Cachannel and PLC/IPtransduction cascades.
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Because of ignoring the basic knowledge training, the medicine graduates lack ba-sic knowledge of clinical pathology, which has seriously affected their improvement of medical skills. The paper analyzes the status of pathology teaching in clinical medicine graduates and the necessity to strengthen clinical pathology general education. Then it proposes measures about enhancing clinical pathology general education, so as to increase the basic clinical pathology knowledge of medical grad-uates.
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Objective To construct the fibroblast-specific non-viral vector pcDNA3-CEP-BMP-2 containing collagen 1A2 enhancer and promoter,and to validate the enhancement of BMP-2 expression in the human dermal fibroblasts by this vector,compared with the routine non-viral BMP2 vector. Methods The sequences for collagen 1A2 enhancer and promotor,and BMP-2 gene were ligated into the pcDNA3 plasmids.The plasmids were transfected into human skin fibroblasts and vein endothelial cells by means of cationic liposomes.The expressions of the plasmids in these two kinds of cells were detected by RT-PCR.The osteogenic phonotypes of fibroblasts were determined.(Results)pcDNA3-CEP-BMP-2,which contained collagen 1A2 enhancer and promoter could enhance the BMP-2 expression in the fibroblasts but not in vein endothelial cells.Osteogenetic phenotypes were more obvious in the fibroblasts transfected with pcDNA3-CEP-BMP-2 than in pcDNA3-BMP-2-transfected ones. Conclusion Collagen 1A2 enhancer and promoter can enhance BMP2 expression in fibroblasts.
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Objective To analyze three different classification criteria, the clinical characteristics of antiphospholipid syndrome(APS)in a cohort of Chinese patients. Methods From January 1996 to October 2006, APS patients diagnosed with different classification criteria were retrospectively studied. Results There were totally 120 APS patients fulfilled at least one criterion, One hundred and one patients fulfilled the 1988 Asherson criteria, 96 patients fulfilled the 1999 Sapporo criteria, and 115 patients fulfilled the 2006 Sydney criteria. The ratio of male to female in a cohort of 115 definite APS patients was 1 to 10.5. The mean period of the disease until entry into the study was 82.6 months, the mean age at study entry was(41?12)years. Ninety patients had thrombosis episodes, among which the most common presenting manifestations were deep venous thrombosis, stroke and skin vasculitis. Forty-six of 92 married women in our cohort had fetal morbidity. Catas- trophic APS occurred in 7 patients. The presence of anticardiolipin antibodies(aCL)was detected in 86 pa- tients, anti-beta-2 glycoproteinⅠantibodies in 58 patients and lupus anticoagulant(LA)in 27 patients. Conclusion The most common presenting manifestations are deep venous thrombosis, stroke and cutaneous manifestations. The sensitivity of Sydney classification criteria is improved by adding anti-beta-2 glycopreteinⅠantibody as one of the laboratory criteria. However, primary APS patients who only presented with thrombo- cytupenia and positive laboratory tests could not satisfy this criterion. In addition, the significance of autoanti- bodies to some coagulant factors in APS needs further study.