Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Peripheral Blood Stem Cell Transplantation ; methods

Hemoglobinuria, Paroxysmal ; therapy ; Humans

Objective To investigate the value of soluble epithelial cadherin(sE-cad)in the differential diagnosis of pleural effusion. Methods Patients were divided into malignant pleural effusion group,infective pleural effusion group and transudation group.sE-cad in pleural fluids obtained during the first thoracocentesis was measured by enzyme-linked immunosorbent assay(ELISA).The concentration of sE-cad in all kinds of pleural effusions was compared.The cut-off value of sE-cad for the differential diagnosis of benign and malignant pleural effusion was determined by ROC curve.The diagnostic value of sE-cad was also compared with common tumor markers such as CEA,CA199,CA125 and NSE.Results The concentration of sE-cad was significant higher in the malignant pleural effusion than in the benign pleural effusion[(38.38?4.15)ng/mL vs(14.17?0.80)ng/mL,P

Carcinoma, Renal Cell ; metabolism ; pathology ; surgery ; Carcinoma, Transitional Cell ; metabolism ; pathology ; surgery ; Carcinosarcoma ; metabolism ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Keratins ; metabolism ; Kidney Neoplasms ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; surgery ; Retrospective Studies ; Survival Rate ; Ureteral Neoplasms ; metabolism ; pathology ; surgery ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery ; Urologic Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

OBJECTIVETo explore the analgesic effect of Zhixin Formula (ZF) and its effects on spinal glial fibrillary acidic protein (glial fibillary acidic protein, GFAP, a marker of astrocyte) , CD11b (a maker of microglia), Toll like receptors (TLR2 and TLR4) and nuclear factor κB (NF-κB) in bone cancer pain model rats.

METHODSTotally 20 male SD rats were randomly divided into the blank control group and the bone cancer pain group, 10 in each group. The bone cancer pain model was induced by injecting ascites tumor fluid containing 3 x 10(3) Walker256 cell line from the left tibia. Ethological tests, X-ray test, and HE staining were performed to confirm a successful modeling. After model was successfully established, 70 male SD rats were randomly divided into seven groups, 10 in each group: the blank control group, the bone cancer pain group (as the model group), the Western medicine (WM) group (Tramadol Hydrochloride), the high dose ZF group, the middle dose ZF group, the low dose ZF group, and the Chinese medicine (CM) group (Wulin Zhitong Capsule). Fourteen days after modeling, rats in the high, middle, and low dose ZF groups were administrated by gastrogavage with 9, 4.5, and 2.25 g/kg ZF water condensed preparation respectively, once a day for seven consecutive days. On day 21 MS typical protein expressions including GFAP, CD11b, TLR (2,4) and NF-κB from cornu dorsal medullae spindis L4-L5 were detected by immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot.

RESULTSCompare with the blank control group, increased weight in the model group was slow and showed a decreasing trend (P < 0.01), spontaneous ambulatory pain score (SAPS) obviously increased (P < 0.01), paw withdrawal threshold (PWT) obviously decreased in the model group (P < 0.05, P < 0.01). Results of lateral tibial X-ray and HE staining showed obvious changes and damage occurred in bone structures of rats in the model group. Immunohistochemistry showed that GFAP expression significantly increased in the model group (P < 0.05). Protein levels of NF-κB also significantly increased in the model group (P < 0.05). Compared with the model group, CD11b expressions obviously decreased in the middle and high dose ZF groups (P < 0.01). Meanwhile, protein expressions of TLR2 and TLR4, as well as NF-κB also obviously decreased (P < 0.05).

CONCLUSIONZF had analgesic effect, which might be probably related to inhibiting proliferation and activation of gliocytes, as well as activation of TLRs and NF-κB.

Animals ; Astrocytes ; Bone Neoplasms ; drug therapy ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Glial Fibrillary Acidic Protein ; Male ; Microglia ; NF-kappa B ; metabolism ; Osteosarcoma ; Pain ; Rats ; Rats, Sprague-Dawley

Objective To analyze the distribution and subcellular localization of NOR1 alternative splice isoforms in human tissues and cell lines. Methods NOR1 open reading frame(ORF) was amplified from human fetal brain cDNA library and subcloned into pCMV/myc vector. The level of NOR1 mRNA in human tissues was determined by real-time RT-PCR. Region spanning exon 2 was amplified from cDNA or genomic DNA by specific primers and sequenced. The expression plasmids of NOR1 were transfected into cells and immunofluoresence assay was performed to determine the subcellular localization of NOR1 protein isoforms in human cells. Results The expression of NOR1 mRNA was high in human adult testis, moderate in human fetus nasopharynx, trachea, brain, and kidney tissues, and weak or undetectable in other tissues. Two splice variants of human NOR1 gene resulted from alternative splicing at exon 2 were identified. Both of isoform 1 and isoform 2 mRNA were detected in human fetus brain. Isoform 2 was the sole isoform in other tissues but brain. Only isoform 2 mRNA was detected in cell lines used in this study, though no genomic deletion of exon 2 could be found in all these cell lines. Immunofluoresence assay showed both isoform 1 and isoform 2 proteins were distributed in cytoplasm. Conclusion Alternative splice isoforms of tumor suppressor gene NOR1 are identified. NOR1 isoform 1 and isoform 2 are both detected in fetus brain. NOR1 isoform 2 lacking of exon 2 is the sole isoform in multiple tissues except for brain. The exon 2 encoded peptide does not affect the subcellular location of NOR1 protein.

Objective To investigate the utility of bedside electromyography (EMG) in diagnosis and management of critical illness patients with suspected neuromuscular diseases.Methods Bedside EMG was performed in 34 critical illness patients with weakness and respiratory involvement,including segmental motor nerve conduction studies,sensory nerve conduction studies,F waves,concentric needle EMG and repetitive nerve stimulation.The clinical manifestations and clinical utility of bedside EMG in critical illness patients with suspected neuromuscular diseases were analyzed. Results EMG was normal in 5 patients.Low-frequency repetitive nerve stimulation showed decrement response of compound muscle action potential (CMAP) in 4 of 8 patients.Motor nerve conduction studies showed CMAP amplitude decreased in 73.3％(22/30) patients,and demyelinating changes was detected in 20.0％ (6/30)patients.F-wave persistence was 0 in 55.0％ (11/20) patients.Amplitude of sensory nerve action potential decreased in 28.6％ (6/21)patients.Bedside EMG could help to confirm or exclude diagnoses and guide the management in 82.4％(28/34) patients,confirm the diagnoses of peripheral neuropathy but have no effect on management in 11.8％ (4/34) patients,and bedside EMG was inconclusive in 2 patients.Conclusions Bedside EMG is useful for the diagnosis and management of critical ill with suspected neuromuscular diseases,while motor nerve conduction studies and repetitive nerve stimulation are more valuable.Individualized protocol for EMG studies should be made on the basis of clinical problem.

Objective: To analyze the association of thrombocytosis with the prognosis of patients with gastric cancer.Methods: The clinical materials of 782 patients with gastric cancer who underwent initial surgery in our hospital between January 1995 and December 1999 were retrospectively analyzed.Kaplan- Meier and Log-Rank test were used to analyze the data.Prognostic factors were analyzed by multivariate Cox proportional hazards model.Results: Thrombocytosis oc-curred in 11.4% (87/782) patients.The platelet level was not significantly different among patients of different gender, tumor stage, and histological differentiation (P>0.05).However, a significant difference was observed in the platelet level among patients with different age and surgical approach (P<0.05).The 1-, 3-, and 5-year survival rates were 75.0%, 40.1% and 28.9% in patients without thrombocytosis and 52.8%, 16.9%, and 13.5% in patients with thrombocytosis (P=0.002).Univari-ate analysis showed that histological differentiation, pathological stage, surgical approach and thrombocytosis significantly affected the survival of patients.While age and gender had no significant impact on patient survival.Multivariate analysis showed that pathological stage, surgical approach, and thrombocytosis were independent prognostic factors for gastric can-cer.The relative risk of death of patients with thrombocytosis was elevated by 1.454 times (RR=1.454, 95% CI: 1.135～1.861, P=0.005).Conclusion: Thrombocytosis is an independent prognostic indicator for the survival of initially treated pa-tients with gastric cancer.

Objective To explore the correlation between the two methods of pulmonary arterial thermodilution technique(ThDCO)and pulse indicator continuous cardiac output(PiCCO)in determining hemodynamic changes in patients undergoing orthotopic liver transplantation(OLT).Methods Eighteen patients with end-stage liver disease(ASAⅡ-IV)were performed OLT and monitored hemodynamic changes by PiCCO method and ThDCO method simultaneously.The hemodynamic parameters including mean arterial pressure(MAP),cardiac output(CO)and systemic vascular resistance(SVR)were measured at different time points before anesthesia induction,after tracheal intubation,after the beginning of operation,at anhepatic phase,at neohepatic phase,at the end of operation,and after operation.Results Good correlation was found between the two methods for the determination of CO and SVR(r=0.987,P0.05).Conclusion The hemodynamics change dramatically in patients undergoing OLT.There is good correlation between the two methods of PiCCO and ThDCO for perioperative hemodynamic monitoring in patients undergone OLT,and PiCCO detection can be used conveniently for real-time continuous monitoring.

To establish a method of directional differentiation and efficient production of neurons from embryonic stem cells (ES cells) in vitro, based on the 4-/4+ protocol described by Bain, a new method was established to induce ES cells differentiating into neurons by means of three-step differentiation using all-trans retinoic acid (ATRA) combined with astrocyte-conditioned medium (ACM) in Vitro. The totipotency of ES cells was identified by observation of cells' morphology and formations of teratoma in immunocompromised mice. The cells' differentiation was evaluated continuously by the detection of the specific cellular markers of neural stem cells, neurons and astrocytes, including nestin, NSE and GFAP using immunohistochemistry assay. The NSE positive cells' ratio of the differentiated cells was determined by flow cytometry. It was found that the transparent circular clusters surrounding embryoid bodies induced with combining induction protocol formed just after 24 h and gradually enlarged later. This phenomenon could not be observed in EBs induced only by ATRA. The NSE positive cells' ratio in the cells induced with ATRA and ACM was higher than that of the cells induced by ATRA at different time points of differentiation, and finally reached up to 73.5% among the total differentiated population. It was concluded that ES cells could be induced into neurons with high purity and yield by means of inducing method combining with ATRA and ACM.
Astrocytes/*cytology ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Neurons/*cytology ; Stem Cells/*cytology ; Tretinoin/pharmacology