1.Study on purification and properties of lactate dehydrogenase from human erythrocyte
Lingxiang MAO ; Ze MENG ; Lining SHI
Chinese Journal of Clinical Laboratory Science 2001;19(1):4-7
Objective To preparing the reference material(RM) of lactate dehydrogenase(LD),we purification of LD from human erythrocyte(RBC)and studied its properties.Methods Using a modified procedure which including complete hemolysis of the RBC,hydroxyapatite treatment,(NH4)2 SO 4 Precipitation,CM- Sephadex C 50,Sephadex G100 and 5'- AMP- Sepharose 4B affinity chromatography.Results The purified LD has a sepecific activity of 163.0 kU/g protein.It is almost free of contaminating enzymes.Two corresponding bands were observed on both PAG plates stained for either protein or LD activity after electrophresis had been done.The apparent Michaelis constants were of 1.000 and 0.179 mmol/L for L-lactate and NAD and of 0.119 mmol/L 0.062 mmol/L for pyruvate and NADH respectively.Conclusion The final purified LD was found to be very similar to that in human serum in catalytic properties.It is intended to be a fundament to prepare a RM for the measurement of LD.
2.Study of Whole Pancreatoduodenal Allotransplantation Model with Enteric Drainage and Portal Venous Drainage in Pigs
Weiming HU ; Fanghai HAN ; Zhaoda ZHANG ; Xiang ZHOU ; Lingxiang MENG
Chinese Journal of Bases and Clinics in General Surgery 2003;0(02):-
Objective To establish the model of pancreatoduodenal allotransplantation in pigs with enteric drainage (ED) and portal venous drainage (PVD). Methods Forty-six hybrid landraces were divided into two groups (donor and recipient groups) randomly, for pancreatoduodenal allotransplantation. Donors were perfused via abdomial aorta without clamping the portal venous outflow with UW solution after heparinization. Whole pancreatoduodenal graft was harvested with segments of abdomial aorta and portal vein and shaped under cold UW solution. Then, the end-to-end anastomosis was performed with the donor iliac artery bifurcation “Y” graft to the recipient superior mesenteric arteries and celiac artery. Furthermore, type Ⅰdiabete model was made by removal of the recipient pancreas. The venous anastomosis was reconstructed between the donor portal vein and the recipient superior mesenteric vein. Meanwhile, the end-to-side anastomosis was performed with the donor common iliac artery bifurcation “Y” graft to the recipient abdomial aorta and the side-to-side intestinal anastomosis was performed between the donor duodenum and the recipient jejunum. External jugular vein was intubated for transfusion. The levels of blood glucose, insulin and glucagon in blood were measured before and during the operation and 1, 3, 5, 7 d after operation. Results Twenty-three cases of pancreatoduodenal allotranplantations were performed on pigs. One died from complication of anesthesia. Success rate of operation was 95.7%.Complications of operation happened in 2 cases in which one was phlebothrombosis, incidence 4.5% and the other was duodenojejunal anastomotic leak, incidence 4.5%. The level of blood glucose increased within 30 min and recovered on the 2nd day after removal of pancreas. The levels of insulin and glucagon decreased within 30 min and recovered on the 2nd day after removal of pancreas. Rejection curred at the 1st day and reached the worst level on the 9th day after transplantation without the change of insulin and glucagon in blood and clinical symptoms of rejection. Conclusion Pancreatoduodenal transplantation in pigs can treat type Ⅰ diabete. ED and PVD can keep the function of endocrine in normal. The techni- que of duodenal transplantation with ED and PVD may pave the way for the further development of pancreas transplantation in clinic.
3.Detection of multifunctional Th1 cells in peripheral blood and pleural effusion in TB patients
Meizhong LI ; Zhengang QIU ; Lingxiang MENG ; Bin SHU ; Qunyi DENG ; Mingxia ZHANG
International Journal of Laboratory Medicine 2014;(15):1971-1973
Objective To investigate the level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cell in peripheral blood and hydro-thorax of the TB patients and its clinical significance .Methods 49 patients with tuberculosis(TB) including 14 cases of tuberculous pleurisy and 27 individuals with latent TB infection were selected and 66 healthy individuals were selected as the controls .PMA and ionomycin were adopted to stimulate mononuclear cells in whole blood and pleural effusion .The secretion status of CD4+ T cells cy-tokines was detected by using the intracellular cytokine staining and the flow cytometric analysis .Results According to the differ-ent cytokines generated by CD4+ T cells ,which were divided into 7 cell subgroups :IL-2+ IFN-γ+ TNF-α+ ,IL-2+ IFN-γ+ ,IL-2+TNF-α+ ,IFN-γ+ TNF-α+ ,IL-2+ ,TNF-α+ and IFN-γ+ cell subgroups .The proportion of peripheral blood IL-2+ IFN-γ+ TNF-α+multifunctional Th1 cells in the TB patients was significantly lower than that in the healthy controls and the individuals with latent TB infection(P<0 .01) ,the expression levels of IL-2+ IFN-γ+ cells and IFN-γ+ TNF-α+ cells were significantly lower than those in the individuals with latent TB infection and the healthy controls (P<0 .05);TNF-α+ cells was higher than that in the healthy con-trols and the individuals with latent TB infection (P<0 .05) .The other subgroups had no obvious change .The response level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cells in the pleural effusion mononuclear cells (PEMC) was higher than that in the peripher-al blood mononuclear cells(P<0 .05);IL-2+ cells in peripheral blood mononuclear cells (PBMC) was lower than that in PEMC (P<0 .01) .Conclusion The response of non-specific Th1 cells is related with the clinical outcome of TB infection ,IL-2+ IFN-γ+TNF-α+ multifunctional Th1 cells plays a certain role in the protective immunoreaction of TB .
4.Application of computer assisted navigation system in endoscopic sinus and skull base surgery.
Lan CHENG ; Rongping CAO ; Guozhen MENG ; Qi HUANG ; Dongming HOU ; Lingxiang HU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2012;26(17):796-798
OBJECTIVE:
To evaluate the applicative value of computer assisted navigation system in endoscopic sinus and skull base surgery.
METHOD:
Forty-nine endoscopic surgery procedures were performed with the help of computer assisted navigation system, among which there were 25 cases of recurrent sinusitis and nasal polyps, 9 cases of nasal and sinus tumour, 7 cases of cerebrospinal fluid rhinorrhea, 2 cases of meningoencephalocele, 4 cases of congenital choanal atresia, 1 case of pituitary tumor and 1 case of foreign body in middle cranial fossa.
RESULT:
The preoperative time was 5-13 minutes for preparation, 7 minutes in average. The target error was less than or equal to 1.5 mm. All the 49 cases had successful surgery without complications.
CONCLUSION
Computer assisted navigation system can help the surgeon to determine the sinus, skull base and adjacent anatomic landmarks correctly, improve surgical accuracy and safety, and reduce intraoperative and postoperative complications.
Adolescent
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Adult
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Aged
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Child
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Child, Preschool
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Computer Systems
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Female
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Humans
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Infant
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Male
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Middle Aged
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Paranasal Sinuses
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surgery
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Skull Base
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surgery
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Surgery, Computer-Assisted
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methods
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Young Adult