The 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase activity of Sclerotinia sclerotiorum is one of the multifunctional enzyme AROM activities, which catalyzes a reversible conversion of shikimate 3-phosphate (S3P) and phosphoenolpyruvate (PEP) to EPSP and inorganic phosphate, and is inhibited by the herbicide glyphosate (N-phosphonomethyl glycine). AROM protein has been purified from Sclerotinia sclerotiorum and the EPSP synthase has been analyzed. The results indicated that the optimal pH and temperature of EPSP synthase were 7.2 and 30℃ respectively. The activation energy of the heat-deactivated reaction of the enzyme was found to be 69.62 kJ/mol. Both of the substrates, S3P and PEP, were showed to inhibit the reaction rate when their concentrations exceeded 1 mmol/L and 2 mmol/L respectively. The Km of 140.98 μmol/L for PEP and 139.58 μmol/L for S3P were obtained by Dalziel equation which was a steadystate kinetic equation of the enzymatic reaction with the double substrates. The kinetic pattern of the enzyme was consistent with a sequential mechanism. Inhibition of the EPSP synthase reaction by glyphosate was competitive with respect to PEP, with the Ki 0. 32 μmol/L, and noncompetitive with regard to S3P. Activation by [ K+ ] was observed in the forward reaction. The Km (PEP) was lowered by increasing [ K+ ], while the Km (S3P) changed irregularly and the Ki (PEP) was enhanced.

Objective To explore the nursing experience of the safe intra-hospital transport of patients with the severe aspiration of the inhalation injury. Method The nursing measures for the intra-hospital transport of 2 cases of smoke pot inhalation injury caused by extracorporeal membrane oxygenation treatment were taken, including disease risk assessment, preparation for transport, organization of a transport team, effective vital signs monitoring during transport, extracorporeal membrane oxygenation (ECMO) pipeline monitoring and nursing and observation and nursing of complications. Result No emergency was found during the transport of patients and both of them were safely transported. Conclusion Such nursing measures as pre-transport assessment and preparation and bettering predictive nursing for the patients with severe inhalation pulmonary injury treated with extracorporeal membrane oxygenation are key to the safety during intra-hospital transport.

Objective　To evaluate whether the stimulation on eyes of bendazac lysine eyedrops could be reduced by cold storage.Methods　 Mate design scheme and double-blind clinical trial were performed. 160 healthy eyes was divided into two groups: the test group was given bendazac lysine eyedrops which was stored in refrigerator at 4℃ over night, and the control group was given it which was stored at room temperature.Results　ln the control group and the test group, the occuring rate of stimulation was 19.30% and 6.25%. The cases of B、C stimulation degrees were 31 and 10; the average stimulation degrees of each group were 1.21 and 1.07, the scoring change for eye stimulation were 1.57±1.50,0.79±1.40,all respectively. The statistical difference was significant (P＜0.05 or P＜0.01).Conclusion　The eye stimulation of bendazac lysine eyedrops can be reduced by cold storage.

Objective To investigate the clinical efficacy and safety of hybrid artificial kidney in the treatment of patients with uremia.Methods 30 maintenance hemodialysis patients who were altered to be treated by hybrid artificial kidney(treatment group)were compared with 29 patients who were treated by hemodialysis only(control group).The amelioration of clinical symptom,remove of urea nitrogen(BUN),creatinine(Cr).Middle Molecular Substance(MMS)and variation of blood regular test were observed.Results BUN,Cr,haemoglobin level,leucocyte count and blood platelets count improved between after and before treatment(P ＜0.05),but there was no significant difference between the two groups(P ＞ 0.05).Clearance rate ofMMS(25.6%)and improvement of clinical symptoms in treatment group were better than those of control group(5.9%)and difference was statistically significant(P ＜0.05,P＜0.01).There was no serious adverse events in two groups.Conclusion Hybrid artificial kidney could remove molecular substances better and markedly improv clinical symptoms.It can make up for the shortcomings of conventional hemodialysis.

Objective To evaluate serum C-reactive protein (CRP) level and its correlation with malnutri-tion,anemia. Methods Thirty-eight elderly and 42 young maintenance hemodialysis patients were studied. Blood samples were collected at the day on hemodialysis for the determination of red blood cell(RBC), hemoglobin(Hgb), hematocrit (Hct), transferrin (TF), serum albumin (Alb), serum creatinine (SCr), blood urea nitrogen (BUN) and CRP; and triceps skinfold (TSF), arm circumference (AC), arm muscle circumference (AMC) were measured. Results The levels of TSF、AC、AMC、RBC、Hgb、Hct、TF、BUN、SCr、Alb in elderly maintenance hemodialysis pa-tients were lower than those of young patients; but serum CRP was higher( P < 0.05 or P < 0.01), The serum-CRP is negatively correlated with hemoglobin(Hgh), hematocrit (Hct) and serum albumin(P <0.01, P <0.01, P <0.05) in elderly maintenance hemodialysis patients. Conclusion Serum CRP is the sensitive index which reflecting early malnutrition and micro-inflammation.

AIM: To explore the pathogenesis of aplastic anemia (AA), we identified the crucial isoform of cyclin D that determine the proliferation of the cord blood CD34~+ cells and observed effects of AA serum on the expression of crucial cyclin D isoform in umbilical cord blood CD34~+ cells. METHODS: The CD34~+ cells were isolated with MIDI-MACS system. The isoforms of cyclin D were detected by RT-PCR and Western blotting. Methylcellulose culture system was used to measure the formation of CFU-GM. The expression level of crucial cyclin D isoform was assayed by RT-PCR and Western blotting after the CD34~+ cells were incubated in AA serum. RESULTS: The crucial cyclin D isoform in CD34~+ cells was cyclin D3. The AA serum inhibited the formation of CFU-GM and down-regulated expression level of the cyclin D3 from the mRNA to protein level, respectively. CONCLUSION: The AA serum inhibits the proliferation of CD34~+ cells and down-regulates level of cyclin D3, this may be one of hematopoiesis inhibition mechanisms in AA.

The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation (BMT) mice was studied. Fifty-six Balb/c mice were divided into 3 groups: normal control group, BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respectively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th,14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells (BMNC) was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed that in ligustrazine treated group the peripheral blood leukocytes, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMT control group (P＜0.01 or P＜0.05). In conclusion, ligustrazine could obviously enhance the CD31expression on the surface of BMNC after syngenic BMT in mice, which may be one of the mechanisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.

The effects of ligustrazine on the expression of LFA-1, ICAM-1 in bone marrow tissue and the mechanism promoting hematopoietic reconstitution following bone marrow transplantation (BMT) were investigated. The 150 mice were randomly divided into 3 groups: normal group, saline group and ligustrazine group. The normal group received no treatment, while in the saline group and ligustrazine group, the mice were subjected to normal saline (0.2 ml, twice a day) and ligustrazine (0.2 ml, twice a day) respectively through a gastric tube. At the 7th, 14th, 21st and 28th day after BMT, survival rate, colony forming unit of spleen (CFU-S), peripheral blood cells and bone marrow mononuclear cells (BMMNC) were measured, histological changes in bone marrow tissue were observed and the expression level of LFA-1, ICAM-1 was detected. In ligustrazine group CFU-S counts on the 10th day and the peripheral blood WBC, PLT, BMMNC counts, hematopoietic tissue volume as well as the expression level of LFA-1 on the 7th, 14th, 21st, 28th day after BMT were higher than in saline group (P<0.01 or P<0.05). Mature RBC volume and the expression level of ICAM-1 were significantly lower in the ligustrazine group than in the saline group (P<0.01 or P<0.05). In the ligustrazine group, fat tissue volume was higher on the 7th, 14th day after BMT (P<0.01) and was lower on the 21st, 28th day (P<0.01) after BMT than in the saline group. It was concluded that Ligustrazine could improve bone marrow microenvironment and promote hematopoietic reconstitution.
Animals ; Bone Marrow Transplantation ; Female ; Hematopoiesis ; drug effects ; Hematopoietic Stem Cells ; drug effects ; metabolism ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Lymphocyte Function-Associated Antigen-1 ; biosynthesis ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; Random Allocation

AIM:To observe the anti-scarring effects and safety of triamcinolone acetonide(TA)-loaded hydrogel sustained-release sheeting on stab incision glaucoma surgery(SIGS)with “one-step tunnel method” in rabbit eyes.METHODS:A total of 48 healthy New Zealand white rabbits were randomly selected and divided into 4 groups(12 rabbits in each group), trabeculectomy(Trab)group, SIGS group, polyvinyl alcohol hydrogel(PVAH)sheeting was implanted under the conjunctiva flap during SIGS(PVAH group), and hydrogel sustained-release sheeting loaded with TA was implanted under the conjunctiva flap during SIGS(TA/PVAH group). On the 1, 2, 3, and 4 wk after surgery, the intraocular pressure, filtering bubble morphology, anterior chamber reaction, and other complications were observed and recorded in each group. Then animals were euthanized, and the surgery area tissues of right eye were taken for pathological tissue paraffin section. Masson staining, picric acid-Sirius rose red staining, as well as α-smooth muscle actin(α-SMA)and fibroblast growth factor 2(FGF2)immunohistochemistry staining was performed on every section. The infiltration of inflammatory cells, proliferation of fibroblasts and synthesis of type I and type III collagen fibers in local tissues were observed. The average positive area ratio of α-SMA and FGF2 antibody immunohistochemical staining in each group was calculated and compared.RESULTS: The TA/PVAH group maintained diffuse and elevated functional filtering blebs, while flat filtering blebs appeared in Trab, SIGS and PVAH groups at 2 wk after surgery. Functional filtering blebs were present in 1 eye(33%), 2 eyes(67%)in the PVAH and TA/PVAH group at 4 wk after surgery, respectively, while the other filtering blebs were flattened. Masson staining showed that the hydrogels in PVAH and TA/PVAH groups did not degrade at 4 wk after surgery. Compared with the Trab and SIGS groups, the filtration passages were more obvious, with less collagen fiber proliferation. Sirius red staining showed that the expression of type I collagen and type III collagen in the TA/PVAH group was less than that in the Trab group, SIGS group and PVAH group at 4 wk after surgery. Immunohistochemical staining showed that the α-SMA expression in the TA/PVAH group was significantly lower than that in the Trab and SIGS groups at 1 wk after surgery(P<0.01). The α-SMA expression was the highest in the Trab and SIGS groups at 2 wk after surgery, while the α-SMA expression in the PHAP and TA/PVAH groups was significantly lower than that in the first two groups(P<0.01). Compared with the Trab group, the expression of FGF2 in the PVAH and TA/PVAH group was significantly increased at 1, 2, 3 and 4 wk after surgery(P<0.05). Compared with the SIGS group, FGF2 expression in the TA/PVAH group was significantly increased at 4 wk after surgery(P<0.05).CONCLUSION:In SIGS surgery of rabbit eyes, implanting hydrogel sustained-release sheeting loaded with TA under conjunctival flap can effectively inhibit the scarring of the filtering bleb, which may be the interaction of the anti-scar effect of TA and the stent function of hydrogel.

The parahippocampal gyrus-orbitofrontal cortex (PHG-OFC) circuit in humans is homologous to the postrhinal cortex (POR)-ventral lateral orbitofrontal cortex (vlOFC) circuit in rodents. Both are associated with visuospatial malfunctions in Alzheimer's disease (AD). However, the underlying mechanisms remain to be elucidated. In this study, we explored the relationship between an impaired POR-vlOFC circuit and visuospatial memory deficits through retrograde tracing and in vivo local field potential recordings in 5XFAD mice, and investigated alterations of the PHG-OFC circuit by multi-domain magnetic resonance imaging (MRI) in patients on the AD spectrum. We demonstrated that an impaired glutamatergic POR-vlOFC circuit resulted in deficient visuospatial memory in 5XFAD mice. Moreover, MRI measurements of the PHG-OFC circuit had an accuracy of 77.33% for the classification of amnestic mild cognitive impairment converters versus non-converters. Thus, the PHG-OFC circuit explains the neuroanatomical basis of visuospatial memory deficits in AD, thereby providing a potential predictor for AD progression and a promising interventional approach for AD.