Objective To investigate the possible role and mechanism of activation of pyroptosis classical pathway and alterations in cell adhesion in calcium-containing kidney stones after the action of high concentration of Ca2+ on HK-2 cells.Methods HK-2 cells were cultured in the presence of different concentrations of CaCl2(0,0.1,0.5,1.0,2.0,4.0 and 8.0 g/L)for 24 hours,and cell counting Kit-8(CCK-8)and flow cytometry were used to determine the optimal treatment concentration.Subsequently,the ultrastructure of renal tubular epithelial cells under high Ca2+ condition was observed by transmission electron microscopy after Ca2+ treatment.DCFH-DA staining was used to detect intracellular reactive oxygen species production,and quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot analysis were performed to examine the expression of pyroptosis-related proteins NLRP3,Caspase-1,gasdermin D(GSDMD),adhesive molecules osteopontin(OPN)and CD44 at mRNA and protein levels after high concentration Ca2+ treatment.The expression levels of pyroptosis-related inflammatory factors interleukin(IL)-1β,IL-18 and adhesive molecule monocyte chemotactic protein-1(MCP-1)were detected by enzyme-linked immunosorbent assay(ELISA)after high Ca2+ stimulation.Results Ca2+ showed cytotoxicity for HK-2 cell growth and can promote apoptosis.The higher the Ca2+ concentration,the more toxicity and apoptosis rate for HK-2 cell growth.High concentration of Ca2+ can promote pyroptosis-like morphological changes in HK-2 cells,including loss of cell membrane integrity,release of contents and numerous intracellular vacuoles.Compared with the control group,the expression levels of ROS were sequentially increased in the 1.0 g/L CaCl2 group and the 2.0 g/L CaCl2 group,and the expression levels of pyroptosis-related genes NLRP3,Caspase-1,GSDMD,and the pyroptosis-associated inflammatory factors IL-1β and IL-18,as well as the adhesion molecules OPN,CD44 and MCP-1 were significantly increased(P＜0.05).Conclusion High Ca2+ treatment can cause oxidative stress damage in HK-2 cells to produce ROS,which activates NLRP3 inflammasome,leads to the activation of the classical pathway of pyroptosis and increase the adhesion of cells,and ultimately leads to the formation of kidney stones.

The pathophysiological process of immune inflammatory response after severe trauma is extremely complex, especially manifested in the dynamic changes. In the physiological response state, the inflammatory and anti-inflammatory conditions are in a dynamic balance. The immune inflammatory response is relatively stable, avoiding excessive inflammatory reactions or immunosuppression and reducing further damage to the body. In the pathological response state, the dynamic balance between inflammatory and anti-inflammatory is broken, and it can also lead to persistent inflammatory-immunosuppression-catabolism syndrome (PICS). As a result, it increases serious complications such as uncontrolled inflammatory reactions, sepsis, multiple organ dysfunction syndrome (MODS), and multiple organ failure (MOF). Current researches on post-traumatic immune inflammatory response have also expanded to the genetic level, indicating that the over-expression of genes and the generation and increase of immune response media are likely to be the key reasons for the disorder of immune inflammatory response. The author reviews the research progress of immune inflammatory response mechanism and related clinical intervention after severe trauma, in order to summarize the previous research results and explore the future research direction.

Translational medicine means that to establish a bridge between fundamental research and clinical medicine through bidirectional translation path in order to promote the translation and application from fundamental research to clinical medicine.The purpose of traditional Chinese medicine (TCM) translational model is to collate and build one multidimensional TCM translational model,rationality carry out the research on TCM translation.Three paths have been confirmed,where are from literature research to clinical verification,from clinical application to clinical verification,and from experiment to clinical verification.In this research,methodology of one path mcntioned above could be established,which is the key technology of translational medicine.Take constipation,insomnia,and post-flu cough as target diseases to investigate the herbal compounds which could be qualified for clinical application based on preliminary work.This research not only provided clinical data to TCM translation,but also established one methodology of TCM translation path--"from experiment to clinical verification".

OBJECTIVE To find out the relationship between mucoepidermoid carcinoma and Human papillomavirus (HPV) infection.METHODS Nested polymerase chain reaction (Nested PCR) was adapted to detect HPV DNA from 51 tissues of mucoepidermoid carcinoma using consensus primers, 9 tissues of chronic parotitis were as control.RESULTS HPV DNA was detected in 37 of total 51 mucoepidermoid carcinoma tissues, the detection rate was up to 72.5%, comparatively, it was 22.2% (2 of 9) in chronic parotitis tissues. The HPV DNA detection rate of mucoepidermoid carcinoma tissues was statistically higher than that of chronic parotitis tissues (χ2=8.517, P=0.004).CONCLUSION The results suggested that the HPV infection may cause mucoepidermoid carcinoma.

Objective To investigate the morphological alteration and the protein expression of caspase -3 in long -term denervated posterior cricoarytenoid muscles (PCAMs)in order to find the appropriate time point of rein‐neration in long -term denervated PCAMs .Methods A total of 45 patients with vocal paralysis were recruited and devided into 3 groups ,the 3~6 months denervation group ,the 6~12 months denervation group ,the 1~2 year denervation group .12 adults served as control group .The morphological alteration was evaluated using HE staining and the change in expression of caspase -3 ,an apoptosis related factor ,were observed using immunehistochemistry stain and western blot .Results With elongation of denervation time ,there were increased denaturation in the mus‐cle fibers .The nucleus moved inside and some of them concentrated .Caspase-3 showed weak staining in innerva‐ted ,however ,by 3-6 months of muscle denervation there was a significant accumulation of caspase -3 protein in myofibers ,6~12 months and 1~2 years of denervation ,expression of caspase -3 protein in myofibers was de‐creased significantly .In western blot ,the change in protein expression of caspase -3 was observed an 21-fold (P< 0 .01) increase from 3~6 months denervated muscles to innervated muscles ,11-fold (P< 0 .01) increase from 6~12 months denervated muscles to innervated muscles ,3~fold (P< 0 .01) increase from 1~2 year denervated muscles to innervated muscles .Conclusion The morphological alterations and changes in expression of caspase -3 indicated there was a high amplitude of apoptosis in denervated posterior cricoarytenoid muscles within 1 year .

OBJECTIVE: To investigate the prevalence of human papillomavirus in nasopharyngeal carcinomas of Fujian province in China. METHOD: Samples from 70 patients with NPC and 25 controls. All samples were detected HPV DNA by polymerase chain reaction (PCR) suing GP5+/6+ and MY09/11 primers and genotyped by surface plasmon resonance (SPR) and HPV 16/18 E6 and LMP-1 using immunohistochemistry and EBER using in situ hybridization. RESULT: Only 2 cases of 70 patients were showed evidence of HPV DNA by PCR, the 2 HPV positive cases subtype HPV-70 and HPV-18 were genotyped by SPR, both the 2 HPV positive cases are non-keratinizing carcinomas (the HPV-70 positive one is differentiated and the HPV-18 positive one is undifferentiated), both the 2 HPV positive cases do not show any evidence of EBV. Data showed that 57 of 70 NPC detected EBER positive, but only 25 out of 70 NPC samples were detected LMP-1 positive. CONCLUSION Our study showed a low prevalence of human papillomavirus in NPC patients of Fujian province in Southern China, there is no evidence about HPV and EBV co-infection.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma ; China ; epidemiology ; DNA, Viral ; isolation & purification ; Epstein-Barr Virus Infections ; epidemiology ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; epidemiology ; virology ; Papillomaviridae ; classification ; isolation & purification ; Papillomavirus Infections ; epidemiology ; Young Adult

OBJECTIVE: To analyse the risk factors involved in local failure following radiotherapy and chemotherapy to nasopharyngeal carcinoma. METHOD: A retrospective analysis was carried out to review the histopathological data from 308 NPC patients who recieved medical treatment between 2004 and 2006. The incidence and the risk factor for local treatment failure were evaluated in a model that included the following factors: sex, age, T and N staging, histological grade of primary tumor, presence of cervical lymph node metastasis, size and laterals of positive neck nodes, levels involved, ways of radiation and condition of concurrent chemotherapy. Univariate chi2 test and multivariate stepwise logistic regression model were used for the analysis. Statistical analysis of survival of patients with local residues and recurrence was performed using Kaplan-Meier method. RESULT: Ninty-three cases (30.2%) presented local residues and recurrence in 308 patients of nasopharyngeal carcinomas. In the univariate analysis, it was confirmed that the following variables correlated to local failure, i. e., T staging (P < 0.01), N staging (P < 0.01), presence of cervical lymph node metastasis (P < 0.05), size and laterals of positive neck nodes (P < 0.05, respectively). In the multivariate analysis, the most ignificant risk factors for local failure were the T staging only. Kaplan-Meier analysis showed that overall survival rates of 71 NPC patients with local residues and recurrence who received re-treatment were 77.2% at 1 year, 40.4% at 3 years, 22.4% at 5 years, respectively. CONCLUSION T staging is the key risk factors in determining the development of local failure following radiotherapy or chemotherapy in NPC patients. Patients with primary tumor infiltrating bone have the higher risk of developing local residues and recurrence. Retreatment to the patients suffering from local failure can imrove survival rates.
Adult ; Aged ; Analysis of Variance ; Bone Neoplasms ; pathology ; Carcinoma ; Female ; Humans ; Lymph Nodes ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; radiotherapy ; Neck ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Neoplasm, Residual ; Retrospective Studies ; Risk Factors ; Survival Rate ; Treatment Failure ; Tumor Burden

OBJECTIVE: To study whether cancer stem cells promotes resistance of laryngeal squamous cancer to irradiation mediated by hypoxia. METHOD: Hep-2 cells were respectively cultured in hypoxia and normoxia environment, and the express of HIF-la was detected by western blot. Then they were radiated with different doses of gamma-rays. After that we detected growth inhibition ratio with MTT assay, cell circle and ratio of CD133+ cells with Flow cytometry at different times. RESULT: MTT assay showed that inhibition ratio of the hypoxia group was lower than that of the normoxia group after different doses of gamma-rays at each time point, and the difference was significant 24 h after 10 Gy irradiation (P < 0.05). The results of Flow cytometry demonstrated that cells of the two groups were arrested at G1 phase, and cells ratio in G1 phase of the hypoxia group was higher than that of he normoxia group after 10 Gy irradiation. The ratio of CD133-positive cells was higher in the hypoxia group than in the normoxia group after radiation, and difference was significant 24 h after 10 Gy irradiation (P < 0.05). In each group, the ratio of CD133-positive cells became higher after radiation than that before radiation (P < 0. 05). CONCLUSION We can conclude that cancer stem cells play an important role in radioresistance mediated by hypoxia.
Carcinoma, Squamous Cell ; radiotherapy ; Cell Hypoxia ; Cell Line, Tumor ; Gamma Rays ; Humans ; Laryngeal Neoplasms ; radiotherapy ; Neoplastic Stem Cells ; cytology ; Radiation Tolerance

Objective To analyze the clinical characteristics, short-term and long-term outcome, overall survival and prognosis of nasal-type,natural killer T-cell lymphoma(NK/TCL) patients. Methods Clinical data of 45 patients with nasal NK/T cell lymphoma from Jan 2001 to Dec 2008 were analyzed retrospectively. Results The complete remission rate was 53.3% (24/45)and the partial remission rate was 17. 8% (8/45), the overall response rate was 71.1% (32/45). At a median follow-up of 15 months, the 1-year,3-year and 5-year survival rates were 68. 3% ,57. 1% ,and 43. 7% respectively. Multivariate analysis showed that B symptoms was independent prognostic factors of NK/TCL( P = 0. 001 ). Conclusions NK/TCL showed specific characteristics, and the long-term survival rate is still poor. Investigation of optional treatment is needed.