Objective The immune magnetic separation(MACS)method of DNT cells was set up and their content Was detected in peripheral blood of healthy man.Methods DNT cells were separated from the peripheral blood.Trypan blue staining and flow cytometry were used to detect their activity and purity,and flow cytometry Was used to detect their content in normal human peripheral blood.Results DNT cell activi ty using MACS sorting was＞97%,the purity Was 82.77%;DNT cells accounted for(6.25±2.61)% (n=45)in the TCRαβ+T cells.Conclusion MACS Can quickly sort out high purity DNT cells,and do not affect the vitality of cells.DNT cells accounted for(6.25±2.61)%in the TCRCD3+T cells.

OBJECTIVE: Using the method of lactate dehydrogenase (LDH) assay, to observe the activities of rat cerebral microvascular endothelial cells (CMECs) intervened by Tongluo Jiunao Injection (TLJNI), a traditional Chinese compound drug removing toxin to dredge brain collaterals, and then further study the effects of different kinds of conditioned mediums (CMECs-CM) of cerebral microvascular endothelial cells on ischemia and ischemia/reperfusion cerebral cortex cells, and to probe into the drug pharmacological mechanisms of CMECs in modulating the neurons. METHODS: Three kinds of CMECs (normal, ischemic and ischemic/reperfusional) were all treated by TLJNI previously, and then the three pairs of CMECs-CM without serum were collected respectively for LDH assay. Rat cerebral cortex neurons were also primarily cultured and then divided into similar three groups (normal, ischemic and ischemic/reperfusional). The neuron responses caused by CMECs-CM at different concentrations were observed by using LDH transudation rate assay. RESULTS: The LDH release values of ischemic and ischemic/reperfusional CMECs with TLJNI treatment were obviously reduced (P<0.01) compared with the same kinds of CMECs untreated. For ischemic neurons, both conditioned medium of ischemic CMECs (Is-CM) and conditioned medium of ischemic CMECs with drug treatment (IsT-CM) in high concentration of 100% increased the LDH transudation rate (P<0.01), while in low concentration of 10%, IsT-CM reduced the transudation rate (P<0.05). For ischemia/reperfusion neurons, all kinds of CMECs-CM reduced the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, 10% or 50% showed relatively stronger effects, and both conditioned medium of normal CMECs (N-CM) group and conditioned medium of ischemic/reperfusional CMECs (Rp-CM) group had statistical significance (P<0.05 or P<0.01). For normal neurons, all kinds of CMECs-CM increased the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, only conditioned medium of normal CMECs (N-CM) had statistical significance (P<0.05 or P<0.01). CONCLUSION: The study shows that TLJNI is capable of preventing the damage of CMECs from both ischemia and ischemia/reperfusion states. Chinese drug can restrain the brain ischemia and ischemia/reperfusion damage by the media that CMECs modulate the neurons, demonstrating the pharmacological mechanisms of TLJNI. This work also indicates that there exist some active substances against ischemia/reperfusion injury secreted from CMECs-CM with TLJNI treatment.

OBJECTIVETo study the changes of metalloproteinases 1, 2, 13 in granulation wound after the treatment of vacuum-assisted closure (VAC).

METHODSThe chronic wounds in 5 patients were treated with VAC. The expressions of MMP-1, 2, 13 in the granulation tissues of the chronic wounds were determined and quantified using RT-PCR technique before and at 1, 4, 7 days after the treatment.

RESULTSThe MMP-1, 13 mRNA showed obvious decrease, with the steepest variation of MMP-13. The MMP-2 mRNA also showed a decreased tendency, though in an undulatory fashion.

CONCLUSIONVAC can promote healing of chronic wounds through depressing the expressions of MMP-1, 2, 13 mRNA and protein synthesis, depressing the degradations of collagen and gelatin.

Collagen ; metabolism ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 13 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Negative-Pressure Wound Therapy ; Polymerase Chain Reaction ; Time Factors ; Wound Healing

BACKGROUNDOlder patients with malignant spinal tumors are difficult to treat because they have many co-morbidities including osteoporosis. The purpose of this research is to discuss the technique and clinical outcome of bone cement enhanced pedicle screw fixation combined with vertebroplasty (the Sandwich Procedure) for elderly patients with severe osteoporosis and malignant spinal tumors.

METHODSThis study includes 28 consecutive elderly patients with malignant thoracic or lumbar spinal tumors. There were nine patients with myelomas, and 19 patients with metastatic bone tumors. The Sandwich Procedure began with curettage of the tumor and a vertebroplasty with bone cement (polymethyl methacrylate, PMMA), followed by PMMA enhanced pedicle screw fixation. Patients were evaluated with the visual analogue scale (VAS), oswestry disability index (ODI), American Spinal Cord Injury Association (ASIA) neurological function classification, and the radiographic degree of kyphosis (Cobb angle). Data were analyzed using paired t-test to compare the pre- and post-operative values. The complications, local recurrences, and the survival status were also recorded.

RESULTSThere was no operative mortality, and the mean operative time was 210 minutes (range 150 - 250 minutes). The average blood loss was 1550 ml (range 650 - 3300 ml). The average amount of cement for vertebroplasty was 3.6 ml (range 3 -5 ml). The VAS, ODI, and ASIA scores were significantly improved after surgery (P < 0.05). However, we found no differences between the pre and post-operative Cobb angles. The shortest survival time was 3 months, and we found no evidence of local recurrence in this group of patients.

CONCLUSIONThe Sandwich Procedure is a safe operation and provides symptomatic relief in these difficult patients, permitting further treatment with chemotherapy or radiotherapy.

Aged ; Bone Cements ; Bone Screws ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Neoplasms ; surgery ; Thoracic Vertebrae ; surgery ; Vertebroplasty ; adverse effects ; methods

OBJECTIVETo observe the specific cellular and humoral immune responses after immunization with recombinant adenovirus Ad5F35-LMP2 in rhesus monkeys.

METHODSSixteen rhesuses were immunized with Ad5F35-LMP2 through intra-muscular injection in three groups: high dosage group (1.5 x 10(10) TCID(50)/rhesus), medium dosage group (1.5 x 10(9)TCID(50)/rhesus), low dosage group (1.5 x 10(8)TCID50/rhesus) and the last group was control (PBS 4 ml/rhesus). They were totally immunized three times at intervals of one month. The EBV-LMP2 specific cellular immune responses were tested during the 0, 4, 8, 12 weeks by Elispot after immunization respectively. And the titers of anti-LMP2 antibody were tested by EIA at the same time.

RESULTSEBV-LMP2 specific cellular and humoral immune responses which were induced by recombinant adenovirus Ad5F35-LMP2 can be found in all the three dosage groups. The potency of immune responses was related with the dosage of immunization. Higher dosage elicited more potent immune response.

CONCLUSIONThe recombinant adenovirus Ad5F35-LMP2 could elicit LMP2 specific cellular and humoral immune responses in rhesus.

Adenoviruses, Human ; genetics ; Animals ; Cell Differentiation ; Herpesvirus 4, Human ; genetics ; Immunity, Cellular ; immunology ; Immunization ; methods ; Macaca mulatta ; Recombinant Fusion Proteins ; genetics ; immunology ; Viral Matrix Proteins ; genetics ; immunology

BACKGROUND/AIMS: To investigate the beneficial effect of N-Acetylcysteine (NAC) on pancreatic microvascular perfusion in acute necrotizing pancreatitis (ANP). METHODS: Fifty-four rats were divided into a control group, an ANP group and an NAC-treated group. The ANP model was established by a retrograde injection of 3% sodium taurocholate into the pancreatic duct. The NAC-treated group received an intravenous infusion of NAC just 2 hours before and 30 minutes after the induction of ANP. The pancreatic microvascular perfusion was measured with laser Doppler flowmetry and pancreatic samples were collected for histological examination. RESULTS: The microvascular perfusion in the NAC-treated group decreased slightly and exhibited a significant increase compared to the ANP group (p<0.01). A pathological examination revealed that edema and inflammatory infiltration decreased, and the hemorrhaging and necrosis of the pancreas were significantly reduced. CONCLUSIONS: NAC could improve pancreatic microvascular perfusion and alleviate the severity of sodium taurocholate-induced ANP, possibly representing a new therapeutic approach to prevent the progression of ANP.
Acetylcysteine ; Animals ; Atrial Natriuretic Factor ; Edema ; Infusions, Intravenous ; Laser-Doppler Flowmetry ; Microcirculation ; Necrosis ; Pancreas ; Pancreatic Ducts ; Pancreatitis, Acute Necrotizing ; Perfusion ; Rats ; Sodium ; Taurocholic Acid

OBJECTIVETo explore the diagnostic methods and reasonable surgical interventions for the chronic obstructive pancreatitis due to the inflammatory lesions at the opening of the pancreatic duct.

METHODSFrom January 2002 to November 2010 the data of 28 patients who were diagnosed as the chronic obstructive pancreatitis (COP) was retrospectively reviewed. Out of the 28 patients, it was analyzed that the clinical manifestations, diagnostic methods, surgical finding and surgical interventions of the 13 patients who were diagnosed as COP due to the inflammatory lesions at the opening of the pancreatic duct in the exploratory operation accompanying recurrent acute abdominal pain with increased serum amylase and lipase, dilation of entire pancreatic duct on imaging before surgery. The conditions included pain recrudescence, quality of life, pancreatic changes on imaging and the serum amylase and lipase after surgery were recorded.

RESULTSAll the 13 patients had clinical manifestations of COP. However, 12 patients had different manifestations on imaging from those chronic pancreatitis imaging due to tumors at the duodenal papilla, ampulla or inner pancreatic duct. Via exploratory operation and magnetic resonance cholangiopancreatography (MRCP), there were short pancreaticobiliary common channel or pancreas divisum existing in most patients. There was no acute abdominal pain with the increased serum amylase and lipase in the 12 patients who receiving the transduodenal mastoid, ampulla and pancreatic ductal opening incision and plasty, the paramastoideus incision and plasty in the visit.

CONCLUSIONSThe imaging character of COP due to the inflammatory lesions at the opening of the pancreatic duct is the dilation of the pancreatic duct without the chronic obstruction in the bile duct. The patients with short pancreaticobiliary common channel or pancreas divisum easily suffer COP due to the stenosis of the pancreatic ductal opening caused by the duodenal mastoiditis or paramastoiditis. The local plasty surgery to correct the stenosis at the pancreatic ductal opening and improve the drainage of the pancreatic duct is an easy and effective management.

Adult ; Aged ; Female ; Humans ; Inflammation ; Male ; Middle Aged ; Pancreatic Ducts ; pathology ; Pancreatitis, Chronic ; diagnosis ; pathology ; surgery ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the relationship between CD4- CD8- T cells ratio and IL-4, IFN-gamma levels in the peripheral blood of patients with pancreatic carcinoma.

METHODSFlow cytometer was used to analyze the CD4- CD8- T cells ratio in the peripheral blood of patients with pancreatic carcinoma and the IL-4, IFN-gamma levels were detected by ELISA.

RESULTSThe ratio of CD4- CD8- T cell in CD3+ T cell from 25 pancreatic carcinoma patients was (4.2 +/- 1.7)%, the ratio of CD4- CD8- T cell in CD3+ T cell from 45 healthy person was (6.3 +/- 2.6)%, there was significant deviation between the two groups (P < 0.01). The IL-4 level of 25 pancreatic carcinoma patients was (86.3 +/- 23.3) fg/L, the IL-4 level of 45 healthy person was (56.2 +/- 9.2) fg/L,there was significant deviation between the two groups (P < 0.01). The IFN-gamma level of 25 pancreatic carcinoma patients was (16.4 +/- 4.8) fg/L before operation, the IFN-gamma level of 45 healthy person was (27.4 +/- 3.8) fg/L, there was significant deviation between the two groups (P < 0.01). The ratio of CD4- CD8- T cell in pancreatic carcinoma patient after operation was higher than before operation. It could be found negative correlation between CD4- CD8- T cells ratio and IL-4 level in pancreatic carcinoma patient,it could also be found positive correlation between CD4- CD8- T cells ratio and IFN-gamma level in pancreatic carcinoma patient. In pancreatic carcinoma patient, the CD4- CD8- T cells ratio and IL-4 level was significant associated with clinical stage (P < 0.05), but no relationship with histological differentiation (P > 0.05), it could be found no relationship between IFN-gamma level and clinical stage, histological differentiation (P > 0.05).

CONCLUSIONThe CD4- CD8- T cells ratio in the peripheral blood of patients is decreased,it may be participate in the carcinogenesis and development of pancreatic carcinoma by influence the IFN-gamma levels.

Adult ; Aged ; CD4-CD8 Ratio ; Case-Control Studies ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; immunology ; T-Lymphocytes ; immunology

OBJECTIVETo study the effect of Platycarya strobilacea Sieb. et Zucc (PSZ) extract on methuosis of human nasopharyngeal carcinoma CNE1 and CNE2 cells and explore the underlying mechanism.

METHODSCNE1 and CNE2 cells were treated with 1 mg/mL PSZ extract and the expressions of Rac1 mRNA and Rac1 protein were detected using RT-qPCR and Western blotting, respectively. Results CNE1 and CNE2 cells showed obvious morphological changes typical of methuosis following treatment with PSZ extract characterized by cell merging, accumulation of large cytoplasmic vacuoles, and membrane rupture without obvious changes in the nuclei. PSZ treatment resulted in up-regulated Rac1 mRNA and Rac1 protein expressions in the cells. Application of EHT 1864 obviously blocked the effect of PSZ extract in inducing methuosis in CNE1 and CNE2 cells.

CONCLUSIONPSZ extract can induce methuosis in CNE1 and CNE2 cells by inducing the overexpression of Rac1.

[Objective]To explore the distribution and expression changes of tight junctional protein JAM-1 in rat models after intracerebral hemorrhage (ICH) and their significance.[Methods]One hundred and twenty-eighty healthy male SD rats were randomly divided into normal control group (n=16) and ICH group (n=112),and the ICH models were induced by stereotactically injecting 75 uL autologous blood into the right caudate nucleus.Seven time points after ICH (6,12,24 and 48 h,and 3,7 and 14 d after ICH,16 rats for each time point) were chosen.BBB permeability was evaluated by Evans blue dye extravasation.The distribution and expression of JAM-1 were detected by immunofluorescence and real-time quantitative PCR.[Results] As compared with that in the normal control group,BBB permeability in the ICH group significantly increased at 24 and 48 h,and 3 and 7 d after ICH (P＜0.05).JAM-1 expression decreased at blood vessels at 12,24 and 48 h after ICH,and JAM-1 expressed at the circulatingleukocytes3 dafterlCH,and abundant JAM-1 positive cells around hematoma were noted in the ED-l-positve macrophages 7 d after ICH.JAM-I mRNA significantly decreased at 12,24 and 48 h after ICH,and significantly increased 7 d after ICH as compared with that in the normal control group (P＜0.05).[Conclusion] JAM-1 experssion changes not only participate in regulation of BBB permeability but also play roles in inflammatory insult after ICH.