Objective To detect the percentage of cultured neuron apoptosis after the neurons were treated with anoxia and specific inhibitors of protein kinase A (PKA) and protein kinase C (PKC). Methods After establishment of the model of neurons cultured under hypoxic condition, the neurons were cocultured with different concentrations of Rp-cAMP and Calphostin C, specific inhibitors of protein kinase A and C, respectively. Then neurons were cultured under an ischemic condition until the number of survived neurons, the activity of mPKA,and mPKC, and the apoptotic neurons stained by TUNEL in each group were observed. Results The activity of mPKA and mPKC significantly increased after the onset of hypoxia. With the increases in concentrations of Rp-cAMP or Calphostin C, the percentage of apoptotic neurons obviously decreased or increased, respectively. Conclusion The pathways of PKA and PKC signal transduction may participate in the hypoxic neuron injury. The functions of these two kinases are opposite for apoptotic regulation. It suggests that the signal transduction of PKA and PKC in hypoxic neurons belongs to a monophasic controlling system and the ratio of PKA to PKC in cells may determine the survival of hypoxic neurons.

BACKGROUND: Botulinum toxin A has been applied to treat muscular convulsion by local injection and shows obvious effects in reducing muscular hypertonia with fewer side effects.OBJECTIVE: To assess the effect of botulinum toxin A on Meige's syndrome concerning facial convulsion, therapeutic effect, and side effects.DESIGN: Randomized self-control clinical study.SETTING: Neurological Department of Southwest Hospital Affiliated to the Third Military Medical University of Chinese PLA.PARTICIPANTS: Between June 2000 and May 2003, 24 outpatients were confirmed of having Meige's syndrome at the Neurological Department of Southwest Hospital, the Third Military Medical University of Chinese PLA and volunteered to receive botulinum toxin A (BTA) injection. Meige's syndrome was presented by convulsive eyelid closure in 10 cases, grinding teeth and smacking movement in 3 cases, involuntary blinking in 2 cases,frowning in 1 case, and mixed manifestations in 8 cases.METHODS: Totally 24 patients with Meige's syndrome were subjected to BTA injection at multiple spots of convulsive muscles by dosage of 2.5-5 U in each spot. Improvement of facial convulsion and curative effects were assessed at least once by clinical examination or telephone follow-up. Convulsion was graded in four degrees according to Cohen's scaling standard.Curative effects: [1] Complete alleviation: Convulsion was reduced to grade 0. [2] Obvious alleviation: Convulsion dropped by more than two grades. [3]Partial alleviation: Convulsion dropped by one grade. [4] Ineffective: Convulsion was not attenuated.MAIN OUTCOME MEASURES: [1] Changes of muscular convulsion intensity after BTA injection in patients with Meige's syndrome; [2] improvement of various symptoms after BTA injection; [3] side effects of BTA.RESULTS: Totally 24 patients entered the result analysis. [1] Before treatment, there were one case of grade 0- Ⅰ, 7 cases of grade Ⅱ, 10 cases of grade Ⅲ and 6 cases of grade Ⅳ, as compared to 16, 6, 2 and 0 cases, respectively, after treatment (χ2 = 95.489 4, P = 7.313 41×10-10). [2] BTA could alleviate eyelid convulsion in an average of 3 days, with curative effect reaching the peak in 2 weeks. Symptoms were found to be completely alleviated in 75% (18/24), obviously alleviated in 17% (4/24) and partially alleviated in 8% (2/24), with the total efficiency of 100%. The effective duration was 8-44 weeks with the average of 24.4 weeks. [3] Muscle weakness at the injected spot and incomplete muscular paralysis at the uninjected spots occurred for a short time after injection, but were alleviated automatically within 1-8 weeks.CONCLUSION: BTA can obviously reduce convulsive degree and enhance the effects in treating Meige's syndrome; moreover, selection of muscles to be injected into and injection spots will help reduce side effects.

Objective To observe the effect of antihypertension treatment at different time after acute onset of stroke on patients' prognosis.Methods Ninety-six patients with ischemic stroke were randomly divided into three groups who received antihypertension treatment separately at the 4th,7th and 10th day after onset.Patients' neurologic deficit and ischemic events were observed at 1 month and 3 months in follow-up.Results Patients who received antihypertension at the 4th day after onset had severer neurologic deficit and more ischemic events than those who received antihypertension at the 7th or 10th day.The patients who received antihypertension treatment at the 7th and 10th day did not show significant difference in prognosis and occurrence rate of ischemic event.Conclusion It is preferable to start antihypertension treatment at the 7th day after onset for patients with ischemic stroke.

Aim To observe the expression of MRTF-A in rat glomerular mesangial cells(GMCs) induced by advanced glycation end products(AGEs) and its effect on ICAM-1 and FN;to explore whether MRTF-A is involved in the process of diabetic nephropathy by affecting NF-κB pathway.Methods Under the condition of AGEs, CCG-1423 and anti-MRTF-A small interfering RNA were used to knock down MRTF-A and MRTF-A plasmid was used to activatt MRTF-A, The expression level of MRTF-A, ICAM-1, FN and p65 in nucleus were detected by Western blot.Results The protein expressions of MRTF-A was increased in AGEs-induced GMCs.The expressions of FN and ICAM-1 and p65 in nucleus were downregulated by knocking down MRTF-A.However, the expressions of FN, ICAM-1 and p65 in nucleus were upregulated by overexpressing MRTF-A.Conclusions AGEs can upregulate the expression of MRTF-A in GMCs, and MRTF-A mediates the protein expressions of FN and ICAM-1 by affecting NF-κB signaling pathway in AGEs-induced GMCs.

Liver ferritin of Sphyrna zygaena(SZLF) with purity of mass spectrum was prepared in batch. Under) the condition of acidifying medium at pH 1.0, PAGE showed that SZLF subunits treated for 20 min began) to dissociate. A whole process of subunit dissociation and recombination was monitored by transmission electron microscopy(TEM). In addition, the changes of size of both protein shell and iron core were also determined) by TEM directly. It was found that in the acid dissociation process of SZLF subunits, the size of iron) core and protein shell showed the same trend of change, which might be related to not only the iron release) of inner iron core but the dissociation and unfolding of the protein shell. The passway of SZLF recombination is a fast step, which is a conversion process from incompact moltenglobule to compact ferritin. Under the assistant of matrix acidity pH 3.0 and laser, SZLF mixed with horse spleen ferritin(HSF) still has capacity to release) its subunits to form subunit ions for mass analysis by a MALDI-TOF mass spectrometer, which indicates that the interaction intensity between the subunits was weaken but they were not unfolded under this pH condition. TEM technology can be applied in studying both dissociation and recombination in ferritin subunits.

Objective To explore the efficacy of intervention combined with traditional Chinese medicine for the treatment of early femoral head necrosis. Methods 62 cases of early femoral head necrosis were treated with interventional and traditional Chinese medicine, all patients were assessed for clinical efficacy before treatment and 6 months after treatment. The clinical hip function, pain index evaluation and rating of the femoral head venography were recorded. Results 41 cases were cured, 18 cases improved,no improvement in 3 cases,the total effective rate was 95. 2%. Clinical hip function after treatment compared with before treatment increased by 32.5%,there was a statistically significant difference (P＜0.01); compared with before treatment, significantly reduced pain (P＜0. 01). After treatment, Ⅲ and Ⅳ were significantly increased number of cases, respectively, compared with the preoperative increase of 146.2% and 160.0%,femoral venography ratings before and after treatment had significant difference(P＜0.01). Conclusion Intervention combined with traditional Chinese medicine treatment could significantly improve the early femoral head necrosis.

AIM: To study the effects of prearuptorin C (Pra-C) on myocardial sarcolemma Na~+, K~+-ATPase activity, myocardial mitochondria Na~+, K~+-ATPase, Ca~ 2+ -ATPase, Mg~ 2+ -ATPase activities and apperent Km and Vmax in spontaneously hypertensive (SHR) and renovascular hypertensive rats (RHR). METHODS: ATPase activity was measured with colourmetric method. Apparent Km and Vmax of both Na~+, K~+-ATPase in myocardial sarcolemma and Ca~ 2+ -ATPase in myocardial mitochondria were calculated according to Lineweaven-Burk double-reciprocal plot method with liner regression. RESULTS: The Vmax of both Na~+, K~+-ATPase in myocardial sarcolemma and Ca~ 2+ -ATPase in myocardial mitochondria were lower in SHR untreated group than that in SD control rats, while Km was higher than that in SD control rats. In RHRs untreated group, only Vmax was decreased, while the Km had no statistically change. Pra-C prevented the reduction of ATPase in amount, but not affected their intrinsic characteristics. CONCLUSIONS: The results suggest that both amounts and affinities to ATP of Na~+, K~+-ATPase and Ca~ 2+ -ATPase were decreased in SHRs, but in RHRs, only amounts of ATPase was decreased, while their affinities to ATP were unchanged. Treatment with Pra-C prevents the decrease in amount of ATPase.

Aim To investigate the expression of G protein-coupled receptor TGR5 and its effects on FN and TGF-β1 expression cultured under high glucose condition in rat glomerular mesangial cells , and then to explore the role of TGR5 in diabetic nephropathy. Methods INT-777 and TGR5 plasmid were used to activate TGR5 under high glucose(HG,30 mmol·L - 1 glucose ) condition, and anti-TGR5 small interfering RNA(TGR5 siRNA) was used to knock down TGR5. The protein expression of FN and TGF-β1 in rat me-sangial cells was detected by Western blot. Results TGR5 could be detected in rat glomerular mesangial cells. Both FN and TGF-β1 protein levels could be in-creased by high glucose compared with control group(P < 0. 05),and be inhibited by activiation of TGR5(P <0. 05). On the other hand,knockdown of TGR5 could increase FN and TGF-β1 protein to abnormal levels(P< 0. 01,P < 0. 05). Conclusion TGR5 suppresses HG-induced FN and TGF-β1 expression in rat glomer-ular mesangial cells,suggesting a protective role in the process of diabetic nephropathy.

AIM: To observe the effect of Tangmaiantai on the plasma lipid and renal function of rat with diabetic nephropathy. METHODS: The rat model of diabetic nephropathy was established by using STZ. The effect of Tangmaiantai on rat blood glucose before meal, serum TC, TG, Ccr, BUN and urinary protein were observed. RESULTS: Tangmainantai could decrease the level of serum TC, TG, Ccr, BUN and urinary protein, comparing to that of the model group (P

Objective To explore the curative effect and safety of CTLA-4Ig transgene for experimental autoimmune encephalomyelitis (EAE). Methods The adenovirus loaded CTLA-4Ig was injected into Wistar rat EAE models through the lateral cerebral ventricle. The clinical symptomatic and electrophysiological changes were observed to judge the efficiency of CTLA-4Ig. Results After injection of AdCTLA-4Ig into the lateral cerebral ventricle of Wistar rat EAE models, the time of EAE onset was delayed and the incidence rate was significantly reduced. And the transductive function tested by brain-stem auditory evoked potential (BAEP) and somatosensory evoked potential (SEP) was obviously improved. Conclusion CTLA-4Ig is effective in managing EAE.