[Objective] To study the correlation between kidney cell DNA degradation and postmortem interval within the span of 6-48 hour after the subject rats′ death.[Methods] To select 18 healthy mature female SD rats and equally divide them into 6 groups.To execute the rats with cervical spine articulation and put the rats under the incubator temperature of 25.1℃ (the average temperature of the 5 previous Decembers in Guangzhou prefecture).Sample kidney tissue from the rat separately 0 hour,6 hours,12 hours,24 hours,36 hours,48 hours,and 60 hours after the rats′ execution to prepare monoplast suspension,which is committed to comet assay.The comet images were captured by fluorescence CCD.Kinetic Comet 4.0 software was used to analyze images.Relevant data were collected by kinetic Comet 4.0 software and were subjected to Kruskal-Wallis test.[Results] Within the postmortem interval of 6-48 h,the number of SD rat kidney cell DNA fragments increased as the postmortem interval lengthens.So did the comet tail length.The Oliver tail moment and tail DNA of comet also showed sign of increase in positive proportion to the postmortem interval (their values corresponding to 60-hour-postmortem-interval were not obtainable.Kruskal-Wallis test indicated:the discrepancies of TL among the 6 groups were all significant (P ＜ 0.01).The difference of TM between 6 h group and 12 h group was not significant (P ＞ 0.05).The difference of TM between 24 h and 36 h was significant (P ＜ 0.05).The difference of TDNA among 6 h,12 h,and 36 h groups were not significant (P ＞ 0.05).The difference of TDNA between 36 h and 48 h was significant (P ＜ 0.05).[Conclusion] Degradation of nuclear DNA of the rat kidney cells increases as the postmortem interval lengthens and comet assay may provide important empirical evidence for determining the postmortem interval.

Objective:To evaluate the prediction value of preoperative Fibrinogen(FIB) in combination with neutrophil-lymphocyte ratio(NLR)for the prognosis of operable pancreatic cancer patients.Methods:The clinical data of 124 patients who underwent radical resection for pancreatic cancer in our hospital from Jan 2010 to Dec 2018 were retrospectively analyzed.The patients were divided into three groups according the high and low NLR, FIB value which defined by the receiver operating characteristic curve (ROC). The clinicopathological data and overall median survival time were compared between the three groups.Results:Univarate analysis showed that age, tumor stage, NLR, FIB and F-NLR score were associated with the prognosis( P<0.05), while multivariate analysis showed that high F-NLR score was the independent prognostic factor. The median survival time of patients with F-NLR scores 0, 1 and 2 group was 30.6, 20.3 and 13.9 months( P<0.05). The prognosis of high F-NLR score was significantly worse than that of low F-NLR score( P<0.05). Conclusions:A high preoperative F-NLR score was a promising predictor for the prognosis of pancreatic cancer patients after radical resection.

Recepteur d′origine Nantais(RON),a tyrosine kinase receptor ,is a growth factor receptor belonging to the proto-oncogene met family and has been proved to display abnormal expres?sion in many types of tumors. The RON receptor is activated by binding to the ligand macrophage stim?ulating protein,overexpression of the receptor,variants of the proto-oncogene and by point mutations of the kinase region. The downstream transduction of RON by mitogen-activated protein kinases and phosphoinositide3-kinase signaling pathways can help regulate the proliferation,apoptosis,migration and invasion of tumor cells. A better understanding of the mechanisms and related signaling pathways of RON activation in tumor progress and development will provide more information for the RON-based target therapy.

BACKGROUND: Both Allergic rhinitis and bronchial asthma are allergic diseases of respiratory tract. They frequently coexist in the same patients.OBJECTIVE: To study the incidence of allergic rhinitis in patients with bronchial asthma in Nanjing, and to analyze the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.DESIGN: The questionnaire survey.SETTING: Department of Otorhinolaryngology-Head and Neck Surgery and Department of Respiratory, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.PARTICIPANTS: A total of 134 patients with bronchial asthma including 73 males and 61 females were selected form Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from February 2001 to April 2005. Their ages ranged from 3 to 72 years. Severity of asthma of all patients was classified based on Global Initiative for Asthma (GINA). The diagnostic criteria of allergic rhinitis were established referred to the guideline of Haikou Meeting (November, 1997). All patients provided the confirmed consent.METHODS: The questionnaire survey of 134 bronchial asthma patients from the city of Nanjing was conducted. Their age, classification, severity, treatment status, and the family history of all 134 patients were investigated in details. Differences of age, age at onset, and course of disease between rhinitis and asthma of all patients were examined by two independent t tests. The correlation between clinical characteristics of allergic rhinitis and bronchial asthma was analyzed by Spearman rank correlation.MAIN OUTCOME MEASURES : Incidence of allergic rhinitis in patients with bronchial asthma and the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.RESULTS：Of all 134 bronchial asthma patients，82(61．2％,82/134)were accompanied with allergic rhinitis．There were 45 males and 37 females．Of 82 patients with allergic rhinitis accompanying with bronchial asthma，severity was classified into grade Ⅰ (mild pause,n=56)，grade Ⅱ (mild persistence，n=21)，grade Ⅲ (moderate persistence，n=4)，and grade Ⅳ (severe persistence，n=1)．In addition,allergic rhinitis was classified into periodicity (n=65) and persistence(n=17)．According to the classification and seventy system recommended in ARIA,63 patients had mild symptoms and 19 had moderate and severe symptoms．Severity of allergic rhinitis was positively related to that of bronchial asthma(r=0.689,P＜0．01)．CONCLUSION: The morbidity of allergic rhinitis combining with bronchial asthma is high in Nanjing, China. While, their classification and severity are highly coherent.

BACKGROUND:Chitosan is a kind of natural biomaterial and is characterized by great biocompatibility, progressive degeneration and absorption and excellent mechanical property; however, whether it may become an ideal cytoskeleton in the engineering of cartilage tissue or not should be researched further.OBJECTIVE: To observe the hydrophilicity and adsorptivity to human nasal septum chondrocytes and the effect of its function of a novel scaffold made by [poly (dl-lactide-co-glycolide)] (PLGA)/chitosan nonwoven cloth embedded with lecithin (LEC) and poly-L-lysine (PLYS).DESIGN: Blank control study.SETTING: Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from October 2005 to June 2006. Chitosan nonwoven cloth was provided by Hainan Xinlong Company. The mainly technical parameters were detailed as the follows: degree of deacetylation ≥ 90% and relative molecular weight 2-5 × 105. PLGA/chitosan nonwoven cloth scaffold was made in Department of Otolaryngology-Hean and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA and High Polymer Institute of Sun Yat-sen University. The mainly technical parameters were detailed as the follows: mole ratio between monome lactide and glycolide 75:25, porosity 82%-86%, pore diamater 100-300 μm, shear strength 48 MPa, depth 1.5 mm and completely degenerated duration 14-18 weeks. Human nasal septum chondrocytes were the septal cartilage of nose which was derived from operated patients with deflection of nasal septum under sterile condition.METHODS: PLGA/chitosan nonwoven cloth was sheared into pieces with the size of 1.5 cm × 1.5 cm, dipped in 0.01 volume fraction of LEC anhydrous alcohol and 1 g/L PLYS for 6 hours, dealt with ultraviolet radiation after dehydration for 1hour, dipped in 0.75 volume fraction of ethanol for 24 hours, washed with Hanks solution, and incubated for 24 hours. After operations mentioned above, two kinds of novel scaffolds containing various components were obtained, and they were simple scaffold and scaffold embedded with LEC and PLYS. Cells derived from the third generation of human nasal septum chondrocytes were used to make suspension. In addition, cell suspension was grown on those two scaffolds to determine the degree of hydrophilicity through observing diffused degree of cell suspension. Whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not were observed under phase contrast microscope so as to determine adsorptivity between cells and scaffolds; meanwhile, growth of cells and production of matrix were also observed.MAIN OUTCOME MEASURES: ① Hydrophilicity of those two kinds of scaffolds (diffused degree of cell suspension of human nasal septum chondrocytes on scaffold) and adsorptivity to cells (whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not); ② growth of cells and production of matrix.RESULTS: ① When simple scaffold was put in cell suspension of human nasal septum chondrocytes, cell suspension showing like balls attached to the surface of scaffold, and then, scattered into space of scaffold gradually. During the period of culture, phase contrast microscope indicated that masses of cells attached to the surface of fiber of scaffold.When the petri dish was shaken, cell groups drifted irregularly. The adherent rate was (21±3.7)%. With the cultured time passing by, matrix was not produced. ② When scaffold embedded with LEC and PLYS was put in cell suspension of human nasal septum chondrocytes, cell suspension scattered into space of scaffold rapidly. The adherent rate was (89±5.6)%, which was higher than that of single scaffold group. This suggested that scaffold showed a strong hydrophilicity.Phase contrast microscope indicated that chondrocytes as the form of monome or community were distributed between scaffold and fiber averagely and attached to the surface of fiber of scaffold. When the petri dish was shaken, cell groups did not drift irregularly, and only a few of cells were scarred at the bottom of petri dish. This suggested that scaffold had a strong adsorptivity to cells. One week after culture, matrix showing like cobweb was produced among fibers of scaffold.With the cultured time passing by, matrix was produced abundantly.CONCLUSION: The novel scaffold of PLGA/chitosan nonwoven cloth embedded with LEC and PLYS is characterized by an excellent hydrophilicity and adsorptivity to human nasal septum chondrocytes; meanwhile, it also can secrete matrix.

Objective To explore the status of mutations of k-ras gene in colorectal cancer (CRC) patients and to make theory preparation for the k-ras mutation detection in diagnosis laboratory. Methods The Genomic DNA was extracted, mutation analysis of k-ras was detected by PCR and bi-direction sequencing in the 56 specimens. Results Rate of k-ras mutation was 46.63 % (26/56) including 76.92 % (20/26) located at codon 12, and 23.08 %(6/26) located at codon 13, and no mutation was found at both codons simultaneously. G>A transition is the most common type of k-ras mutation,GGT>GAT (G12D) is the predominant mutation at codon 12 and GGOGAC (G13D) at codonl3. Chi-square analysis revealed the k-ras mutation was significantly correlated to the gender of the patients. Conclusion The k-ras mutation is mainly located at the codon 12, G>A transition is the most type of k-ras mutation in CRC. k-ras mutation seems to correlate with the gender of CRC patients.

Family Health ; Humans ; Paraganglioma ; diagnosis

Objective To investigate the basic biological functions of NKX3.1 with construction of a series of eukaryotic expression vector of NKX3.1 to study its effects on cell cycle and apoptosis.Methods The full-length reading frames (ORF) of NKX3.1 was amplified from normal human prostate tissue with polymerase chain reaction (PCR),the ORF was cloned into T vector,and then NKX3.1 was cloned into eukaryotic expression vector pcDNA3.1 (+) and pcDNA3.1 (-) with this T vector as template.NKX3.1 in pEGFP C1 was transfected into PC3 cell line,and C1 pEFP was taken as control to observe the localization of NKX3.1 in cells.At the same time,NKX3.1 in pcDNA3.1 and pcDNA3.1 were transfected into PC3 cells and the cells were collected after 24 ～48 hours to detect cell cycle and apoptosis by flow cytometry.Results The eukaryotic expression vector of NKX3.1 was constructed successfully.Fluorescent protein was ocated in the nucleus after transfection of NKX3.1 in pEGFP C1,but pEGP C1 was both expressed in nucleus and cytoplasm.There was significant effect on the early apoptosis (P＜0.01),but no effect on the cell cycle (P ＞ 0.05) after transfection of NKX3.1 compared to PC3 cell-transfected with empty vector of pcDNA3.1.Conclusions The result data suggest that NKX3.1 be nuclear protein and have effect on the early apoptosis of PC3 cell line based on the construction of eukaryotic expression vector.

Objective To explore the changes in inflammatory reactions in tail-suspension mice infected by Klebsiella pneumoniae from spaceflight.Methods Tail suspension was used to simulate the physiological effects of microgravity.C57BL/6 mice were randomly divided into control (Con),control+K.pneumoniae T16-169 (Con+T16-169),tail suspension (TS) and tail suspension+K.pneumoniae T16-169 (TS+T16-169) groups.The level of inflammatory cytokines TNF-α,IL-6 and IL-1β mRNA in lung tissue and the plasma cytokine concentration were detected by RT-qPCR and xMAP technology,and HE staining was used to represent the morphological changes in lung tissue.Results Compared with the control group,the expression of inflammatory cytokines in lung tissue and plasma concentrations of all experimental groups were increased,and the difference in TS+T16-169 group was the most significant (P<0.01 or P<0.001).HE staining showed that the lung tissues in Con+T16-169 and TS+T16-169 groups were damaged in different degrees,and the damage of TS+T16-169 group was the most serious.Conclusion The K.pneumoniae from spaceflight significantly increases the expression of inflammatory cytokines in lung tissue and plasma concentrations after infecting tail-suspension mice,and induces more serious damages to the lung tissue,which suggests that inflammatory reactions can be increased in tail suspension mice infected by K.pneumoniae from spaceflight.

Objective To analyze the efficacy and safety of different chemotherapy regimens for treatment of progressive patients with small cell lung cancer (SCLC) brain metastasis after radiotherapy. Methods 96 SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy were divided into four groups: carmustine group (Group A, 28 cases), temozolomide group (Group B, 19 cases), topotecan group (Group C, 24 cases) and no chemotherapy group (Group D, 25 cases). Results In terms of brain metastases, there were no complete response cases in the whole groups. The rates of partial remission (PR), stable disease (SD) and progression of disease (PD) in Group A were 17.8%(5/28), 53.6%(15/28) and 28.6 % (8/28), respectively, the response rate (RR) of intracranial lesions was 17.9 % (5/28), and disease control (CR+PR+SD) rate was 71.4%(20/28). The rates of PR, SD and PD in Group B were 15.8%(3/19), 63.2 % (12/19) and 21.1 % (4/19), respectively, the RR of intracranial lesions was 15.8 % (3/19), and disease control rate was 78.9 % (15/19). The rates of PR, SD and PD in Group D were 8.3 % (2/24), 54.2 %(13/24) and 37.5 % (9/24), respectively, the RR rate of intracranial lesions was 8.3 % (2/24), and disease control rate was 62.5 % (15/24). In Group D, there was no response case, and 20 patients with PD (80.0 %) were found. The median progression-free survivals (PFSs) were (3.64 ±0.43) months, (4.68 ±0.49) months,(3.58 ±0.50) months, (2.60 ±0.31) months in Group A, B, C and D, respectively, and the median overall survivals (OSs) were (18.80±1.74) months, (18.76±1.85) months, (19.10±1.64) months and (9.64±0.84) months, respectively. The median OS of Group A, B or C was longer than that of Group D (P=0.002). The differences of grade Ⅲ-Ⅳhematologic toxicities among the four subgroups were not statistically different. Patients in Group B had better tolerance to nausea and vomit. In Group D, the central nervous system symptoms such as fatigue and headache occurred frequently. Conclusions The response rate and OS of SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy are improved after chemotherapy, however, PFS is not significantly prolonged. The efficacies of carmustine, temozolomide and topotecan are similar in short and long term, besides, temozolomide shows less adverse events and a higher disease control rate. The application of chemotherapy that could penetrate the blood-brain barrier can improve the efficacy on SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy with well tolerance.