Objective] To summarize the clinical experience of Professor Yu Ying’ao in treatment of epilepsy.[Method]Starting from Professor Yu Ying’ao epilepsy treatment of two typical cases, and discuss the treatment ideas and methods.[Results] Professor Yu Ying’ao is good at treating epilepsy, many cases, he believes that phlegm and distraught as the main disease epilepsy mechanism. Emphasizes the importance in the pathogenesis of phlegm, that Qianzhen Zhi Xian in the treatment of epilepsy through usage, and treatment based on syndrome differentiation. Treatment of multiple in Puji Benshi Fang Baijin pill on the basis of adding Huotan Kaiqiao, Zhenxin Pinggan Anshen, Xifeng Zhijing. Often the effect is very good.[Conclusion] Professor Yu Yingao’s experience in the treatment of epilepsy is unique, it is worth learning.

Objective To investigate effect of guanosine diphosphate (GDP) on the mitochondrial respiratory oxygen consumption and the mitochondrial membrane potential (MMP) of rat brain and explore the relationship of the change of uncoupling proteins (UCPS) activity with the oxygen consumption and MMP. Methods The mitochondria of rat brain were isolated by centrifugation. Mitochondria oxidative respiratory consumption was measured by Clark electrode after the treatment of GDP at different concentrations so as to calculate mitochondrial state 3 respiration (ST3), mitochondrial state 4 respiration (ST4), respiratory control rate (RCR), and the rate of oxidative phosphorylation (OPR). MMP was detected by Rhodamine 123 method at the different concentrations of GDP. Results With the increase of GDP concentration form 0 to 1.0 mmol/L, the values of ST3, ST4 and OPR were reduced while RCR was elevated. But when the concentration increased to 1.4 mmol/L, the former 3 indexes begun to increase while the later declined. When the GDP concentration reached to 1 mmol/L, the inhibitory rate was only 35.1%, 51.3%, 14.2% to ST3, ST4 and OPR respectively, while RCR was increased to 133.2%. No matter the concentration was over 1 mmol/L or under 1 mmol/L, the ability of inhibition was attenuated. MMP reached to the highest point when GDP exerted the highest inhibitory rate on mitochondrial respiratory oxygen consumption. Conclusion GDP, an inhibitor of UCPS, can regulate the respiratory oxygen consumption and MMP of the isolated rat brain mitochondrial directly in a dose-effect fashion. The change of UCPS activity can affect the respiratory oxygen consumption and MMP.

AIM:To observe the effect of GDP on uncoupling proteins(UCPs) activity and the efficiency of oxidative phosphorylation in hypoxic exposed rat brain mitochondria.METHODS: Adult SD rats were randomly divided into three groups (control, acute hypoxia and chronic hypoxia groups). The animals were placed into a hypobaric chamber simulated 5 000 m high altitude for 0, 3 and 30 d, respectively. The mitochondria from rat brain were isolated by centrifugation. The activity of UCPs was detected by the method of [H3]-GTP binding with UCPs specifically. The maximal binding content (Bmax) and the dissociation constant (Kd) were determined by Scatchard plot. The mitochondrial potential was measured by rhodamine 123 method. Oxidative respiratory consumption was measured by Clark electrode. The experiments were conducted under the conditions with or without GDP (1 mmol/L), respectively. RESULTS: For exposed to hypoxia, Bmax and the oxidative consumption of uncoupling respiration were increased. Kd, MMP and RCR were decreased. UCPs activity was inhibited by GDP in three groups. Kd was increased 61.01%, 83.13% and 71.52% and Bmax was decreased 23.18%, 35.20% and 33.38%, respectively. The values in the acute hypoxic group were changed markedly. The sensitivity of UCPs to GDP was elevated significantly by hypoxia. With the reducing of UCPs activity, oxidative consumption of uncoupling respiration was decreased whereas RCR and MMP were increased. The results elucidated increase in the efficiency of oxidative phosphorylation.CONCLUSION: GDP increases the mitochondrial membrane potential and decreases the oxygen consumption of uncoupling respiration in hypoxic exposed rat brain mitochondria by inhibiting UCPs activity. The results suggest that the change in UCPs activity is one of the factors of mitochondrial dysfunction in oxidative phosphorylation induced by hypoxia.

Objective To investigate the pathologic characteristics of skip metastases of mediastinal lymph node in lung cancer. Methods Of 398 lung cancer patients who underwent radical resection and extensive lymph node dissection. 47 patients were diagnosed as with skip N 2 metastases, and their mediastinal lymph nodes were analyzed. Results The skip N 2 metastases of lung cancer appeared as single group or multi-group. The commonest metastatic site was the 7th group lymph node (28.4%), then the 4th, 3rd and 5th group, rating 24.2%, 15.8% and 10.5%, respectively. For cases with squamous cell carcinoma or adenocarcinoma, no any skip N 2 metastasis occurred for the tumor less than 1 cm in diameter, for tumor greater than 1 cm in diameter, the skip N 2 metastasis increased with tumor size. The incidence skip N 2 metastasis of low-grade differentiated adenocarcinoma was significantly higher than high-grade differentiated adenocarcinoma (P

Objective: To investigate the effect of electroacupuncture (EA) on cognitive function in D-galactose (D-gal)-induced aging rats, and the correlation between the effect and nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3)-ASC-Caspase-1 signaling pathway. Methods: Forty-six male Sprague-Dawley (SD) rats were randomly divided into a control group (n=10), a model group (n=12), an EA-7 d group (n=12) and an EA-21 d group (n=12). Except the control group, the other three groups received 42 consecutive days of intraperitoneal injection of D-gal to establish aging rat models with cognitive dysfunction. The control group received the same amount of normal saline via intraperitoneal injection. Two EA groups were given EA therapy for 21 consecutive days (began from the 22nd day of modeling) or 7 consecutive days (began from the 36th day of modeling) accordingly at Dazhui (GV 14), Baihui (GV 20), Shenshu (BL 23) and Zusanli (ST 36). After modeling/ intervention, all four groups received behavioral evaluations by Morris water maze (MWM) test, novel object recognition (NOR) test and step-down passive avoidance (SDPA) test followed by the Western blot (WB) detection of the expression levels of hippocampal NLRP3 inflammasome-associated proteins NLRP3, ASC and Caspase-1. Results: MWM (place navigation test, PNT) results showed that the escape latency in the model group was significantly longer than that in the other three groups (P<0.05), and there was no significant difference among the other three groups on the 1st day of the test (P>0.05). From the 2nd day to the 4th day of the test, there was no significant difference between the EA-21 d group and the control group (P>0.05) in the escape latency; the escape latency was significantly shorter in the EA-21 d group than in the model group and the EA-7 d group (P<0.05). MWM (spatial probe test, SPT) results showed that the time spent in the target quadrant was significantly shorter and platform crossover number was significantly lower in the model group than in the other three groups (P<0.05). The time spent in the target quadrant was longer in the EA-7 d group than in the model group (P<0.05), but was shorter than that in the control group and the EA-21 d group (P<0.05). There was no significant difference in the swimming speed among the four groups (P>0.05). NOR results showed that there was no significant difference in the recognition ratio between the EA-7 d group and the EA-21 d group (P>0.05), and the recognition ratio was significantly higher in the two EA groups than in the model group (P<0.05), but was lower than in the control group (P<0.05). SDPA results showed that the electric shock number was higher in the model group than in the other three groups (P<0.05), and the differences among the other three groups were statistically insignificant (P>0.05). The model group had the shortest step-down latency, followed by the EA-7 d group, the EA-21 d group and the control group in order (P<0.05). The WB results indicated that the expression level of NLRP3 was significantly lower in the control group and the EA-21 d group than in the model group and the EA-7 d group (P<0.05). The expression levels of ASC and Caspase-1 were significantly higher in the model group than in the other three groups (P<0.05), and there was no significant difference among these three groups (P>0.05). Conclusion: NLRP3 inflammasome may be involved in the development of cognitive decline in aging rats; 7 consecutive days of EA intervention can partially improve the cognitive impairment in aging rats though the effect is rather limited; 21 consecutive days of EA intervention may improve the learning and memory abilities in aging rats via downregulating the expression levels of NLRP3 inflammasome-associated proteins in hippocampus.

AIM:To elucidate whether ZFP580 is involved in the cardioprotective effects of hypoxic precondi-tioning (HPC) against hypoxia-reoxygenation (H/R) injury in H9c2 myocardial cells.METHODS: Rat heart-derived H9c2 cells were cultured in DMEM .H/R was induced by incubation under ischemic hypoxia for 3 h and reoxygenation for 2 h.HPC was induced by exposing the H 9c2 cells to 10 min of hypoxia and 20 min of reoxygenation for 3 cycles before H/R treatment.MTT staining and LDH leakage detection were used to evaluate the effects of HPC .Western blotting was used to detect the protein levels of ZFP580, phosphorylated ERK1/2 and cleaved caspased-3.The effects of ZFP580 overexpre-ssion or knockdown on H/R induced apoptosis were determined .RESULTS:The results of MTT staining and LDH leakage detection showed evidence of HPC cytoprotection against H /R-induced cell death in H9c2 cells.ZFP580 protein level and ERK1/2 phosphorylation were significantly increased in the HPC group compared with control group and H /R group. PD98059, an inhibitor of ERK1/2 phosphorylation , significantly suppressed the HPC-induced up-regulation of ZFP580 pro-tein expression.ZFP580 overexpression significantly inhibited apoptosis and caspase-3 activation in H9c2 cells.CON-CLUSION:HPC exhibits cytoprotection against H/R and leads to high level of ZFP 580 protein in H9c2 cells.ZFP580 is regulated by ERK1/2 activation and mediates the anti-apoptotic effect of the ERK1/2 signaling pathway in HPC cytoprotec-tion.

Objective To evaluate the value and clinical application of cardiac structure Z scores in children with preoperative tetralogy of Fallot (TOF) by echocardiography. Methods Considering the result of 31 patients who underwent echocardiography in Shenzhen Children’s Hospital before TOF repair from January 2011 to September 2013, the median age was 10 months (range:3 days to 14 years). Other 31 normal children were selected as normal control group with age-and sex-matched, the median age was 10 months (range: 6 days to 13 years). Echocardiographic measurements included the diameter of right ventricular outflow tract (RVOT), pulmonary valve diameter (PVD), left pulmonary artery diameter (LPAD), right pulmonary artery diameter (RPAD), pulmonary valve peak velocity (PV-Vmax), tricuspid annulus diameter (TVD), mitral annulus diameter (MAD), left ventricular end-diastolic volume (LVEDV) and left ventricular mass (LVmass). Shenzhen Children’s Hospital pediatric echocardiography normal reference values and the regression equation are applied to obtain the forecast average, and using the Z-score formula into the standard value (Z=[M-y]/ MSE ). Results There was no significant difference in body surface area between the TOF group and control group (P>0.05). Compared with control group, the true value of RVOT reduced significantly in TOF group, its Z-score showed negative values increase (-4.87±1.86 vs 0.33±0.93, t=-13.90, P<0.05). PVD, LPAD and RPAD reduced significantly in TOF group, those Z scores showed a negative value increase (-4.15±2.93 vs 0.11±0.85, t=-7.76;-1.35±2.38 vs 0.66±1.17, t=-4.24;-1.47±2.03 vs 0.59±1.04, t=-4.94;all P<0.05). While PV-Vmax increased significantly, and it’s Z score showed a positive value increase (8.38±0.19 vs 0.24±0.98, t=29.50, P<0.05). TVD of TOF patients was slightly larger than the control group (P=0.10), but the Z score was significant greater than the control group (1.00±1.17 vs 0.29±0.52, t=3.06, P<0.05). MV-D, LVEDV and LVmass of TOF patients were slightly decreased than the control group (all P>0.05), but the Z scores of TOF were significantly reduced than the control group (0.25±0.78 vs 0.76±0.65, t=-2.75;-0.92±1.94 vs 0.03±1.01, t=-2.41;-0.83±1.59 vs 0.67±0.69, t=-4.71; all P<0.05). The Z scores of RVOT, PVD, LPAD and RPAD had no significant difference between transannular patch surgery and pulmonary valve-sparing repair patients (-4.89±2.03 vs -4.84±1.67, t=-0.08; -4.73±3.49 vs -3.34±1.75, t=-1.31; -1.88±2.54 vs -0.62±2.00, t=-1.49;-1.89±1.90 vs-0.84±2.15, t=-1.41;all P>0.05). For the Z scores of PV-Vmax, there were no significant difference between transannular patch surgery and pulmonary valve-sparing repair (8.58±1.20 vs 8.12±1.16, t=1.07, P>0.05). Conclusions The Z scores value of cardiac structure can be used in quantitative analysis of RVOT obstruction and development of left ventricular backward of TOF. The Z score of transannular patch surgery is more lower than pulmonary valve-sparing repair patients. The normalized Z score by body surface area correction is important for the preoperative diagnosis and the surgical planning.

ObjectiveTo examine the effects of proprioceptive neuromuscular facilitation(PNF) techniques on lower limb motor function of strokes and its mechanism.MethodsWe used simple random sampling and cross-section survey design. PNF Contract Relax Agonist Contract(PNF-CRAC) techniques were applied to 44 stroke patients. Surface electromyography values(sEMG) was recorded from rectus femoris and hamstring of stroke patients with both low limbs. ResultsPNF-CRAC techniques not only caused the irradiation of muscles activities in the contralateral extremity during unilateral exercise, but also increased agonist EMG activities and the motor control of knee joint in strokes.ConclusionPNF-CRAC techniques can improve the knee stability and enhance the recovery of motor function in paretic lower limb.

Objective:Toprepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) adenovirus, and to investigate their effects on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The microsphere was constructed by encapsulating recombinant adenovirus containing TIMP-1 in biodegradable PELA. The diameter of the microsphere, quantity of virus encapsulated, loading rate, and releasing kinetics were measured. HepG2 cells were infected with the microspheres; the infection efficiency was examined by fluorescent microscope; and the ultrastructure was observed by TEM. The expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR, and the proliferation of HepG2 cells was detected by MTT assay. Results: The microsphere encapsulating recombinant TIMP-1 adenovirus was successfully constructed, with its diameter, entrapment efficiency, and virus loading rate being 1.965, 60.0%, and 10.5×10~8/mg, respectively. About 60% of the viruses were released within 120 h, and the total releasing time was longer than 240 h. Infection with rAdTIMP-1 PELA microsphere efficiently induced TIMP-1 expression in HepG2 cells, and significantly inhibited the proliferation of HepG2 cells, with the inhibitory rate being 47%. Conclusion: PELA microsphere encapsulating recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for the combining macromolecular chemistry and gene therapy for treatment of hepatocellular carcinoma.

Objective To investigate the effects of transforming growth factorβ1(TGFβ1)and β3 (TGFβ3)gene transfer on MMP-2,MMP-9 and TIMP-1 expression in hepatic stellate cells(HSC-T6).Methods TGFβ1 and TGFβ3 expression plagmids were constructed.The recombinant expression plasmid pcDNA3.1(+)-=TGFβ1 and pcDNA3.1(+).TGFβ3 were transfected or cotransfected into HSC-T6.At 24,48 and 72 h after transfection,the expression of MMP-2,MMP-9 and TIMP-1 mRNA were detected by real-time quantitative PCR,and the expression of MMP-2,MMP-9 and TIMP-1 protein were detected by Western blot.The recombinant expression plasmid pcDNA3.1(+).TGFβ1 was transfected into HSC-T6,and positive clones were selected by G418.The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+).TGFβ1,and the expression of MMP-2,MMP-9 and TIMP-1 were detected at 48 h after transfection.Results After transfection with peDNA3.1-TGFβ1,MMP-2 and TIMP-1 increaged remarkably in HSC-T6 cells(P＜0.05),but MMP-9 remained at the sanle level;After transfection with pcDNA3.1-TGFβ3,expression levels of MMP-2,MMP-9 and TIMP-1 mRNA were not changed,but TIMP-1 protein increased remarkably(P＜0.05);in cotransfection group,the expression of MMP-2 was higher than that in the blank and the control groups(P＜0.05),but MMP-9 level was not changed and TIMP-1was decreased compared with that in the TGF-β1 transfection group(P＜0.05).After TGFβ3was transfected into positive clones,the change of MMP-2 wag not significant(P＞0.05).but MMP-9 increaged and TIMP-1 decreased significantly at 48 h after transfection(P＜0.05).Conclusions TGFB3 may inhibit liver fibrosis by increase the activity of MMP-2 and MMP-9,and decrease the activity of TIMP-1.