OBJECTIVE: To establish a RP-HPLC method for the simultaneous determination of Protocatechuic acid and Protocatechuic aldehyde in Fuxuekang granules. METHODS: The chromatography was carried on Achirom Bond-1 C18 column(250 mm?4.6 mm,5 ?m). The mobile phase was composed of methanol - water (20∶80,pH=2.80 adjusted by glacial acetic acid) at a flow rate of 1.0 mL?min-1. The detection wavelengths were programmed at 256 nm for Protocatechuic acid and at 280 nm for Protocatechuic aldehyde. RESULTS: The linear ranges for Protocatechuic acid and Protocatechuic aldehyde were 5.49～73.22 ?g?mL-1(r=0.999 9) and 4.81～64.13 ?g?mL-1(r=0.999 9), respectively, and their average recoveries were 96.86%(RSD=2.52%,n=9) and 97.55%(RSD=3.82%,n=9) respectively. CONCLUSION: The method is simple, sensitive, and reproducible, and it is applicable for the quality control of Fuxuekang granules.

Objective To investigate the expression of Golgi protein?73(GP73) and Ki?67 antigen in gallbladder carcinoma ,and to analyze their correlations with proliferation ,invasion ,and prognosis of gallbladder carcinoma. Methods Streptavid?in?peroxidase (SP) immunohistochemistry was used to detect the expression of GP73 and Ki?67 in surgically resected specimens of 58 gallbladder carcinomas ,15 gallbladder adenomas and 15 gallbladder polyps samples . Results The positive rates of GP73 and Ki?67 protein in gallbladder carcinomas were 72.4% and67.24%,respectively ,which wer significantly higher than those in gallbladder adenomas(GP73:40.0%,Ki?67:26.7%,P<0.05)and in gallbladder polyps(GP73:13.3%,Ki?67:25.0%,P<0.05).The expression of GP73 was positively correlated with tumor differentiation ,Nevin staging and lymph node metastasis(P < 0.05), and the expression of GP73 was positively correlated with tumor differentiation ,Nevin staging(P < 0.05). GP73 expression was positively correlated with Ki?67 expression in gallbladder carcinoma (r = 0.473 ,P = 0.000). Patients with negative expression of GP73 and Ki?67 had longer survival time than those with positive expression of GP73 and Ki?67. Conclusion The expression of GP73 and Ki?67 was associated with proliferation ,invasion of gallbladder carcinoma. The combined detection of GP73 and Ki?67 is conducive to judging the progress and prognosis of the gallbladder carcinoma .

OBJECTIVE:To establish a method for content determination of cyproheptadine hydrochloride in fuyan cream.METHODS:Ultraviolet spectophotometry was applied to determine the contents of cyprohepatadine hydrochloride as well as its adjuvant as two coexisting components without isolation and extraction,the wavelengths were286nm and258nm res_ pectively,linear regression and simultaneous equations were used to analyze the results.RESULTS:Cyproheptadine hydrochlor_ ide and its adjuvant showed good linear relationship in the range of2.52～25.2and2.5022～25.022?g/ml respectively,the average recovery of cyproheptadine hydrochloride was98.61%(RSD=1.08%).CONCLUSION:The present method is simple,convenient,reproducible and reliable,which is suitable for rapid determination for fuyan cream.

Measurements of the magnetic properties and total contents of Cu, Cd, Pb and Fe in 30 automobile emission particulate samples indicated the presence of magnetic particles in them. The values of frequency dependent susceptibility (chi(fd)) showed the absence of superparamagnetic (SP) grains in the samples. The IRM(20 mT) (isothermal remanent magnetization at 20 mT) being linearly proportional to SIRM (saturation isothermal remanent magnetization) (R(2)=0.901), suggested that ferrimagnetic minerals were responsible for the magnetic properties of automobile emission particulates. The average contents of Cu, Cd, Pb and Fe in automobile emission particulates were 95.83, 22.14, 30.58 and 34727.31 mg/kg, respectively. Significant positive correlations exist between the magnetic parameters and the contents of Pb, Cu and Fe. The magnetic parameters of automobile emission particulates reflecting concentration of magnetic particles increased linearly with increase of Pb and Cu content, showed that the magnetic measurement could be used as a preliminary index for detection of Pb and Cu pollution.
Algorithms ; Environmental Monitoring ; methods ; Feasibility Studies ; Magnetics ; Metals, Heavy ; analysis ; chemistry ; Statistics as Topic ; Vehicle Emissions ; analysis

Objective To investigate the relationship between cathepsin L and apoptosis cell in rats after cerebral ischemia reperfusion.Methods Sixty healthy male Sprague-Dawley Rats (10-12 weeks old,260-300 g) were chosen.Based on the random number table method,the rats were randomly divided into sham-operated control group (Sham group,n =10),ischemia-reperfusion group (model group,n =25),and Z-FY-DMK intervention group (CLI group,n =25).Rats were randomly divided into 6 h,12 h,24 h,and 48 h four subgroups in model group and CLI group,respectively.Modified transient middle cerebral artery occlusion was made as Longa described,the intervention groups were injected intracerebroventricularly Z-FY-DMK (20 μg / 1μ1 ×5 μl) preoperative 30 min prior to surgery,Sham group and schemia reperfusion injury (IRI) group were injected intracerebroventricularly dimethyl sulfoxide (DMSO) 5 μ1 (10ml/L) at the same time.Cell apoptosis was detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) straining.Western blotting was used to detect the expression of cathepsin L and caspase-3.Results In the cortical area of ischemic brain,apoptosis cells of sham operation group were rare,while apoptosis of nerve cells of model group with 6 hours reperfusion were visible,and were gradually increased in the order of 12 hours,24 hours and 48 hours.At the same time point,the apoptosis cells of CL intervention group (6 h,12 h,24 h,48 h) were obviously less than model group (P ＜0.05).Western blotting found little visible cathepsin L protein expression in ischemic cerebral cortex preoptic in the sham group.For model group,the cathepsin L expression initially increased in sub groups with 6 hours reperfusion,reached to a peak in sub groups with 12 hours and 24 hours,and remained a high level in sub groups with 48 hours reperfusion.Compared to model group,the cathepsin L expressions of CL intervention group were obviously decreased at all time points (P ＜ O.05).Conclusions Cathepsin L may be involved in neuronal apoptosis by means of caspases 3 pathway.

Objective To study the expression and the clinical significance of hypoxia-induced factor-1α (HIF-1α) in gallbladder cancer tissues,and the role and mechanism of HIF-1α in metformin-suppressed metastasis in gallbladder carcinoma GBC-SD cells.Methods 24 specimens of gallbladder cancer tissues and 5 specimens of chronic cholecystitis were collected from the First Affiliated Hospital of Zhengzhou University between June 2016 and February 2017.Immunohistochemistry and qPCR were used to detect the expression of HIF-1α in gallbladder cancer tissues,in adjacent non-cancer tissues and in chronic cholecystitis,and the clinical significance was analyzed.The model of metastasis was induced by hypoxia;the wound healing assay and the Transwell assay were used to detect the ability of cell metastasis;the expressions of HIF-1α and VEGF in gallbladder carcinoma GBC-SD cells were detected by western blotting assay and immunofiuorescence.Results The expression of HIF-1α in gallbladder cancer tissues was higher than the adjacent non-cancer tissues and in chronic cholecystitis.The expression of HIF-1α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues (P ＜ 0.05).The wound healing rate after 48 h in the negative control group and in the treatment with hypoxia group (1％ O2) in GBC-SD cells were (46.5 ± 4.8) ％ and (67.3 ± 4.0) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group and in the treatment with hypoxia group GBC-SD cells were (147.4 ± 11.7) and (234.4 ± 17.7),respectively.When compared with the negative control group,treatment with hypoxia significantly increased the ability of metastasis and up-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (40.6 ± 7.1) ％,(16.4 ± 9.4) ％,(69.5 ± 4.0) ％ and (22.4 ± 7.4) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (148.4 ± 6.9),(90.0 ± 8.4),(185.8 ± 10.2) and (113.4± 8.6),respectively.When comparcd with the hypoxia group,treatment with metformin and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (43.4 ±4.4)％,(25.9 ±9.0)％,(63.3 ±2.2)％,(46.2 ±4.5)％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (144.2 ± 12.6),(80.2 ±7.7),(203.8 ±7.0),(124.0 ± 5.2),respectively.When compared with the hypoxia group,treatment with HIF-1α inhibitor 2MeoE2 and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).Conclusions The expression of HIF-1 α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues.Treatment with hypoxia significantly increased the expression of HIF-1α and VEGF and promoted metastasis of GBC-SD cells,while treatment with metformin decreased the ability of metastasis induced by hypoxia via inhibiting the HIF-1o/VEGF pathway in GBC-SD cells.

Aim To observe the effects of tetrahydro-biopterin ( BH4 ) on nitric oxide ( NO ) production in the kidney of type 2 diabetic nephropathy ( DN) mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in-to DN phase were divided into 2 groups:DAHP group, subjected to intraperitoneal injection of 150 mg·kg-1 DAHP (n=8);DN group, subjected to intraperitone-al injection of same dose of normal saline containing 5% DMSO ( n = 6 ) . The age-matched db/m mice ( NS group) were subjected to intraperitoneal injection of same dose of normal saline containing 5% DMSO ( n =6 ) . Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u-rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 24h urine volume, 24h urine pro-tein, BH4 , iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser-um creatinine, urine volume, urine protein, BH4 , iN-OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over-production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.

Objective: To investigate the effects of Kindlin-2 on malignant phenotypes of human gallbladder cancer cells and discuss the mechanisms. Methods: The expression level of Kindlin-2 in 30 cases of gallbladder cancer tissues and adjacent non-tumoral tissues collected from the First Affiliated Hospital of Zhengzhou University between September 2012 and May 2013 was assessed by real-time PCR and immunohistochemistry.Lentivirus-mediated Kindlin-2 overexpression was used in gallbladder cancer cell lines GBC-SD and SGC-996.Transwell assay and adhesion assay were investigated to explore the functional role of Kindlin-2 on gallbladder cancer cells.Western Blot was used to test the protein change of epithelial-mesenchymal transition(EMT) characteristics. The t-test was used to analyzed results. Results: The RNA and protein levels of Kindlin-2 in gallbladder cancer tissues were higher than in the non-tumoral tissues (t=4.372, P=0.001; t=7.477, P=0.000). The expression level of Kindlin-2 in gallbladder cancer tissues was correlated with Nevin stage(χ²=5.932, P=0.035). Compared with control groups, the cell-matrix adhesion ability of GBC-SD and SGC-996 with Kindlin-2 overexpression was obviously promoted(1.66±0.03 vs. 1.07±0.22, t=2.710, P=0.041; 2.66±0.24 vs. 1.03±0.02, t=6.610, P=0.020). The number of GBC-SD and SGC-996 cells with Kindlin-2 overexpression passing through the Transwell chamber matrix increased significantly compared with the control groups(116.1±13.9 vs. 54.7±8.4, t=3.781, P=0.019; 136.3±7.5 vs. 64.3±6.4, t=7.302, P=0.002). The wound healing rate of GBC-SD with Kindlin-2 overexpression at 12-hour and 24-hour was higher than that of the group ((42.9±2.2)% vs. (29.7±1.7)%, t=4.690, P=0.009; (65.0±2.4)% vs.(40.4±2.0)%, t=7.945, P=0.001). The wound healing rate of SGC-996 with Kindlin-2 overexpression at 12-hour and 24-hour was also higher than that of the group ((32.9±1.3)% vs. (24.1±1.5)%, t=4.518, P=0.011; (51.3±1.1)% vs. (39.2±1.1)%, t=8.001, P=0.001). The characteristics of EMT were induced in gallbladder cancer cells with Kindlin-2 overexpression, including the up-regulation of N-cadherin, Vemintin and the down-regulation of E-cadherin. Conclusion The expression of Kindlin-2 is up-regulated in gallbladder cancer tissues and Kindlin-2 promoted the malignant phenotypes of gallbladder cancer cells partially by epithelial-mesenchymal transition.

Objective:To evaluate EEG biofeedback therapy on clinical efficacy of children with different subtypes of attention deficit hyperactivity disorder (ADHD) using the integrated visual and auditory integration continuous performance test (IVA-CPT).Methods:Children with ADHD who completed more than 60 times of EEG biofeedback training in Hangzhou Seventh People's Hospital from July 2018 to September 2020 were selected as the research subjects. According to the results of IAV-CPT before treatment, all the children were divided into three subtypes: attention deficit type ( n=21), impulse hyperactivity type ( n=11), and mixed type ( n=30). The differences of symptom improvement between the three subtypes before and after training were compared by SPSS 23.0 software. Results:(1) In terms of response control (a total of 9 items), there was no statistical significance in the attention deficit group before and after treatment (all P>0.05). In the hyperactivity group, the scores of visual and auditory response control, full scale response control, auditory prudence and auditory consistency, visual focus quotient and visual consistency before treatment were significantly lower than those after treatment(all P<0.05). In the mixed group, the scores of visual and auditory response control, full scale response control, auditory prudence, visual consistency, and visual prudence before treatment were significantly lower than those after treatment(all P<0.05). In terms of attention (a total of 9 items), the scores of auditory vigilance, auditory attention, visual attention, full scale attention, visual vigilance and visual speed in the attention deficit group before treatment were significantly lower than those after treatment(all P<0.05). The scores of visual attention, full scale attention, visual focus quotient (all P<0.01), auditory attention and visual speed (all P<0.05) in hyperactivity group before treatment were significantly lower than those after treatment. In the mixed group, the scores of the other 8 items before treatment were significantly lower than those after treatment (all P<0.01, auditory focus quotient were P<0.05), except that the auditory speed had no statistical significance before and after treatment.(2)After EEG biofeedback training, the changes of IVA-CPT scores of the three groups before and after treatment(visual response control (-2.76±24.39), (19.55±19.94), (12.93±25.30), F=3.932, P=0.025), (full scale response control (2.38±20.77), (21.27±15.86), (15.43±25.69), F=3.158, P=0.050), (full scale attention (18.43±27.44), (11.36±11.40), (26.23±18.41), F=4.692, P=0.016), (auditory vigilance (20.23±42.65), (6.55±10.20), (33.63±36.30), F=7.160, P=0.002), (visual vigilance (19.48±28.55), (5.27±10.62), (33.27±28.26), F=10.876, P<0.001), (visual focus quotient (-2.24±23.67), (14.45±13.79), (12.83±21.91), F=3.669, P=0.031) were statistically significant. After LSD comparison, the changes of visual control and total control scores in the attention deficit group before and after treatment were significantly lower than those in the impulse hyperactivity group (all P<0.05) and the mixed group (all P<0.05). In the three items of total attention score, auditory vigilance and visual vigilance, the changes of impulsivity hyperactivity group before and after treatment were significantly lower than those of mixed group ( P=0.050, P<0.05, P<0.01, respectively). The changes of visual attention quotient in the attention deficit group before and after treatment were significantly lower than those in the impulse hyperactivity group and the mixed group (all P<0.05). Conclusion:EEG biofeedback has clinical efficacy in the three groups of subtypes of ADHD, but the efficacy is different, and individualized EEG training programs should be developed for different subtypes of children on the basis of standard TBR therapy protocol.

Objective To study the relation between aquaporin 4 (AQP4) expression in the perihematomal tissue and changes of MRI indicators after intracerebral hemorrhage of rats, and explore the relationship between AQP4 expression and formation of hemorrhagic brain edema. Methods Forty-five male SD rats were randomized into sham-operated group (n=15) and hemorrhage group (n=30). The rats of these groups were equally subdivided into the 1', 2nd, 3rd, 5th and 7th d measurement groups, respectively. The models of intracerebral hemorrhage were established by infusing collagenase into globus pallidus of the rats. MRI was performed 1, 2, 3, 5 and 7 d after the success of model making;edema volume around the hematoma and signal intensity ratio of T1WI, T2WI, and FLAIR sequences in the edema zone were measured and calculated. The rats were sacrificed at the corresponding time point after imaging. Immunohistochemistry staining was performed to observe the expression of AQP4 at each time point. Results The AQP4 expression level of perihematomal tissue in the hemorrhage group was obviously higher than that in the sham-operated group (P＜0.05). Liner positive correlation between the AQP4 expression level and the volume of cerebral edema around the hematoma was noted (r=0.687,P＜0.05). Liner positive correlations between the AQP4 expression level and both signal intensity ratio of T2WI and FLAIR sequences in the cerebral edema region were also found (r=0.640, 0.662; P＜0.05).Conclusion AQP4 has a close relation with the formation and expansion of hemorrhagic cerebral edema; over-expression of AQP4 may promote the formation of edema after intracerebral hemorrhage.