Objective This study investigated a method to rapidly inactivate tumor cells on surgical instruments intraoperatively.Methods Tumor cells were collected by immersing and washing the surgical instruments in 37 ℃ saline.The precipitation was collected by low speed centrifugation and then was cultured to harvest the tumor cells.The tumor cells were immersed in saline and distilled water of different temperatures for different duration of time.Inverted microscopy was used to investigate the changes in morphology.Results After immersion in 55 ℃ distilled water for 60 seconds,the tumor cells were swollen,the cell membranes disappeared,the sizes of the nuclei were reduced,the chromatin was condensed,and some cells lysed and separated from each other.Additionally,these tumor cells floated in the culture medium and lacked any living cells adhering to the walls of the bottle.In the group of tumor cells treated with 55 ℃ saline for 60 seconds,there were no obvious morphological changes of the tumor cell or nucleus.Conclusion The intraoperative immersion of surgical instruments in 55 ℃ distilled water for 60 seconds could completely inactivate tumor cells.

Objective To investigate the relationship between positive peritoneal exfoliated cancer cells and the clinicopathological features of patients with hepatocellular carcinoma before any invasive treatment.Methods Of the 92 patients with hepatocellular carcinoma who underwent laparotomy,ascites fluid was collected in the patients with peritoneal ascites; and peritoneal lavage fluid was collected in those patients without peritoneal ascites.Then,shedded cancer cells in these fluid samples were detected.Results The positive rates of peritoneal cancer cells were associated with the TNM stage,tumor location and tumor size.The positive detection rate of cancer cells in TNM stage Ⅲ and Ⅳ was significantly higher than stage Ⅰ and Ⅱ (38.1％ vs 8.0％ ; P =0.0005).The positive detection rate was higher in tumors located closer to the surface (P =0.0 002),and with larger diameter (P =0.00 007).Conclusion Peritoneal cancer cells were significantly correlated with tumor stage,tumor location and size in hepatocellular carcinoma.

Objective: To obtain monoclonal antibodies against putative protein X4 of SARS-CoV for further study of the structure and function of protein X4. Methods: Balb/c mice were immunized with recombinant Protein X4, hybridoma cell lines secreting monoclonal antibodies against protein X4 were screened by regular cell fusion and subcloning approach. The specificities of these monoclonal antibodies were determined by Western blotting and Immunofluorescecence assay. Results: Three hybridoma cell lines (8A5, 8H6 and 4A2) stable in secreting specific monoclonal antibodies were successfully obtained. They could bind specifically to protein X4 proved by Western blotting. All of them belonged to the IgG2a isotype proved by antigen mediated ELISA. Indirect Immunofluorescence assay indicated that they could specifically bind to protein X4 expressed in SARS-CoV infected Vero E6 cells. Conclusion: Monoclonal antibodies of high specificity against protein X4 have been successfully prepared, which laid the foundation for the further study of protein X4.

Objective：Our aim was to investigate the proportion of autosomal recessive (AR) inheritance among families with patients with Duchenne muscular dystrophy (DMD) and clinical feature in patients with AR form of DMD. Methods:A total of 193 families was studied， 8 of them with at least one girl with “DMD - like” phenotype and 185 with only boys with this kind of phenotype. Based on the number of families with at least one affected girl and the number of patients per sibship among these pedigrees, the proportion of families with DMD inherited as an AR trait was estimated. The clinical examination, family history and serum creatine-kinase were studied in 11 patients diagnosed as AR form of DMD. Results: The proportion of families with AR form of DMD was estimated as 9.4%. The average age of being able to walk is (1.47±1.00) year, serum creatine-kinase levels were (2785.10±1500.29) U/L. The clinical symptom occurred at the average age of (8.11±4.32) year in patients with AR form of DMD. Conclusion： The AR form of muscular dystrophy and DMD not be distingushed clinically. Some families with only affected boys diagnosed as typical DMD, in fact, have the AR form of the disease. This study is very useful for genetic consulting.

Purpose To compare radiation dose and image quality of different scan modes for CT pulmonary angiography (CTPA) including high-pitch flash mode, dual energy CT (DECT) mode, 128-slice mode of dual source CT and 64-slice CT mode. Materials and Methods One hundred and sixty-seven patients with suspected pulmonary embolism were retrospectively studied. All the patients underwent CTPA. Twenty patients were selected randomly from the patients scanned by high-pitch flash mode. Patients who were scanned by the other three modes were selected with body mass index and age matched those in high-pitch flash mode, with 20 patients in each group. Two radiologists assessed signal-to-noise ratio (SNR) and image quality with 5-piont scale. Dose parameters of volume CT dose index (CTDIvol), dose length product (DLP), and effective dose (ED) were compared among the four groups. Results Mean CTDIvol, DLP and ED were (3.72±0.74) mGy, (137.5±28.7) mGy · cm, and (2.34±0.41) mSv for Flash mode;(5.31±1.21) mGy, (181.6±34.5) mGy· cm and (3.24±0.57) mSv for DECT mode;(5.66±1.47) mGy, (198.7±42.1) mGy·cm and (3.58±0.63) mSv for 128-CT mode;and (6.75±1.68) mGy, (231.5±54.2) mGy·cm and (4.21±0.89) mSv for 64-CT mode. There was no significant difference of SNR and image quality among the four modes (P>0.05). Conclusion There are no significant difference of image quality among the four groups. Flash mode allows for minimum radiation dose compared to other modes. DECT mode and 128-CT mode get higher radiation dose with no difference between them. 64-CT mode gets the highest radiation dose.

Objective To detect the DMD gene mutation sites and the regions of breakpoints in Duchenne/Becker muscular dystrophy (DMD/BMD) patients in northern China. Methods Multiplex amplifiable probe hybridization (MLPA) was used to detect the mutation in 59 cases (51 cases with DMD and 8 with BMD) from northern China and dystrophin gene mutations in their parents. Results From northern China and dystrophin gene mutations 59 families found gene deletions in 33 cases of 59 DMD/BMD patients (55.9%), duplications in 6 cases (10. 2%) and point mutation in one case (1.7%). Intron 44 was most frequently affected (n = 13, 33.3%), followed by intron 50 (n = 11, 28.2%) and intron 45 (n=8, 20.5%). The novel mutations were identified, in two patients including two independent duplications carried by patient D1 149 and a point mutation [5208del(A)] carried by patient D1 65, which were not included in Leiden database. In addition, an exon 22 deletion was found in one patient, which was the first reported case in Chinese patients. Conclusions Deletions are mostly located in the hotspot between exon 45 and 50. Duplications mostly occurred in the 5' end of the gene. Intron 44 is the most frequently affected breakpoint in northern Chinese population.

Objective: To study the expression and localization of apoptosis-related protein TFAR19 in TF-1 cells undergoing apoptosis. Methods: Using monoclonal antibody against TFAR19, the expression level and cell localization of TFAR19 were examined by fluorescence microscope, confocal laser scan microscope(CLSM) and flow cytometry. Simultaneously, we also analyzed the relationship of TFAR19 protein with phosphatidylserine (PS) externalization and cell nuclear DNA fragmentation. Results: The level of TFAR19 proteins expressed in TF-1 cells treated with GM-CSF withdrawal was significantly increased compared with normal TF-1 cells, then translocated rapidly from cytoplasm to the nucleus of cells. Appearance of TFAR19 in the nucleus of apoptotic cells preceded the detection of PS externalization and DNA fragmentation. Conclusion: Nuclear translocation of TFAR19 protein is one of the earliest events of cell apoptotic process. These data provided a new clue to further approach to the biological function of TFAR19 and study of cell apoptosis.

Objective: To study the expression of PDCD5 mRNA and its significance in esophageal cancer in Xinjiang Kazakh and Han nationality. Methods: RT-PCR was used to detect the mRNA of PDCD5 in 40 cases of esophageal cancer (18 cases of Kazakh, 22 cases of Han). Results: The positive rate of PDCD5 mRNA expression in 40 samples of esopha-geal cancer tissue, adjacent tissue, and normal tissue was 80.0% (32/40), 80.0% (32/40) and 87.5% (35/40), respectively, with no significant difference (P>0.05). The Ods of cancer tissues, adjacent tissues, and normal tissues were 0.7644± 0.1444, 0.9341 ±0.1631 and 1.8703±0.4767, respectively. The expression of PDCD5 was significantly increased in cancer tissues compared with that in normal tissues (P<0.05). The expression level of PDCD5 mRNA was not significantly correlat-ed with the degree of differentiation and lymph node metastasis. Conclusion: No significant difference was found in PDCD5 mRNA expression in esophageal cancer between Xinjiang kazakh and Han nationality (P>0.05). The expression of PDCD5 is not correlated with the degree of differentiation, depth of invasion or lymph node metastasis. Detection of PDCD5 mRNA expression in esophageal cancer tissues may provide valuable information for patient prognosis.

A new apoptosis-related gene TFAR19 was recently cloned. A primary functional analysis indicates its role in the apoptotic process of TF-1 cells. To clarify TFAR19 was involved in which apoptotic pathway, the changes in TFAR19 expression were observed during the apoptotic processes of Jurkat cells induced with various methods: serum deprivation, VP-16 treatment and Fas McAb activation. Then TFAR19 expression in Jurkat cells was detected with flow cytometry and Western blot, and TFAR19 mRNA with RT-PCR. Our results showed that expression of TFAR19 in Jurkat cells was increased at 12 hours after serum deprivation, 2 hours after VP-16 treatment and 2 hours after Fas McAb activation, respectively. These observations suggested that TFAR19 was involved in the apoptotic process of Jurkat cells induced with serum deprivation, DNA destruction and death receptor activation. It might take effect in the early apoptotic process. TFAR19 is a participant of the "final common pathway" in the apoptotic pathway.

OBJECTIVETo investigate the therapeutic effect of nerve growth factor (NGF) on changes of myelin basic protein (MBP) and functional repair of sensory and motor nerve following sciatic nerve injury.

METHODSThe sciatic nerves of rats were injured by sectioning with shaver,and divided into 3 groups: NGF group (Group A), group of normal saline solution (Group B), untreated group (Group C). The time point of observation was at the 4th week after operation. Sensory evoked potential (SEP) and motor evoked potential (MEP) were detected by Model WD-4000 nerve potential working diagnosis system. Immunohistochemical analysis was used for identification of MBP.

RESULTSThe latency of SEP in the Group A at the 4th week after operation was shorter than that in the Group B (P<0.05). The MEP was elicited in 76% of the Group A and was higher than that in the Group B. Results of immunohistochemistry showed that there were less MBP-positive cells in the Group A than in the Group B in one and four weeks respectively.

CONCLUSIONSNGF can improve the conductive function of injured peripheral nerve and facilitate regeneration of nerve.

Animals ; Evoked Potentials ; Female ; Immunohistochemistry ; Myelin Basic Protein ; metabolism ; Nerve Growth Factor ; pharmacology ; Peripheral Nerve Injuries ; Peripheral Nerves ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; injuries ; metabolism