OBJECTIVE:To establish a method for the simultaneous determination of curdione,germacrone and furanodiene of zedoray turmeric oil in Ruxiankang injection. METHODS:HPLC was performed on the column of Zorbax SB-C18 with mobile phase of acetonitrile-water (gradient elution) at a flow rate of 1.0 ml/min,detection wavelength was 216 nm,column temperature was 30℃,and the injection volume was 10 μl. RESULTS:The linear range was 60-480 μg/ml for curdione(r=0.999 0),40-320 μg/ml for germacrone(r=0.999 0)and 40-320 μg/ml(r=0.999 0);RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries were 95.21%-99.89%(RSD=1.6%,n=6),102.33%-104.89%(RSD=1.0%,n=6)and 97.38%-99.06%(RSD=0.7%,n=6),respectively. CONCLUSIONS:The method is simple and accurate,and can be used for simultaneous contents deter-mination of curdione,germacrone and furanodiene of zedoray turmeric oil in Ruxiankang injection.

Objective To establish a method for the determination of berberine hydrochloride and phellodendrine in Lishukang Capsules by HPLC. Methods Waters Symmetry C18 column (4.6 mm× 250 mm, 5μm) was used, the mobile phase was acetonitrile-0.1%phosphoric acid (0.1 g sodidum dodecyl sulfonate in 100 mL water), with the flow rate of 1 mL/min by gradient elution. The detection wavelength was set at 284 nm. Results Berberine hydrochloride showed good linear relationship (r 2=0.999 1) in the range of 403.4-10 084.7 μg, the average recovery rate was 99.8%(RSD=1.7%). Phellodendrine showed good linear relationship (r 2=0.999 2) in the range of 169.137 6-4228.44 μg, the average recovery rate was 95.5%(RSD=3.2%). Conclusion The method is specific, accurate and convenient to be used for the quality control of Lishukang Capsules.

AIM:A new method for the overall quality control of Radix Scutellariae has been established by HPLC digitized fingerprint. METHODS: The chromatographic fingerprints were obtained from injecting 5 ?L of the sample solution each time on a CenturySIL C_(18) BDS column(200 mm?4.6 mm,5 ?m) with the gradient elution solvent system composed of 1% acetic acid water and 2% acetic acid acetonitrile.The flow rate was 1 mL/min,the column temperature was maintained at(30?0.15)(?C) and the detection wavelength was set at 280 nm.The chromatographic fingerprints were evaluated by the software of the digitized evaluation system of Traditional Chinese Medicine fingerprint with the super information characteristics. RESULTS: 30 co-possessing peaks were selected as the fingerprint peaks of Radix Scutellariae and the similarities between the chromatographic fingerprints and the herbal drugs were calculated by taking baicalin peak as the referential peak.The chromatographic fingerprints were also evaluated by the chromatographic fingerprint index(F). CONCLUSION: This method with good precision and reproducibility can be applied to the quality control of Radix Scutellariae.

Objective To establish an HPLC method for determining the content of swertiamarin in Gantaishu capsules. Methods Using a Waters Symmetry C18 column (4.6 mm×250 mm, 5μm), the mobile phase was acetonitrile-0.02%phosphoric acid, with flow rate of 1.0 mL/min. The detection wavelength was at 250 nm, the column temperature was room temperature. Results The content of swertiamarin showed a good linear relationship in the range of 0.373 556-2.988 448 μg, r 2=1.000 0. The average recovery was 106.81%, and RSD was 3.7%. Conclusion This method is fast, accurate and simple with good reproducibility, which can be applied to the quality control of Gantaishu capsules.

Objective To investigate the changing characteristics of myocardial movement in patients with dilated cardiomyopathy by three?di?mensional speckle tracking echocardiography(3D?STE). Methods The peak systolic global longitudinal train(GLS),global radial strain(GRS), global circumferential stain(GCS)and global area strain(GAS)of left ventricle were measured by 3D?STE technology in 69 patients with dilated cardiomyopathy. According to the left ventricular ejection fraction(LVEF),all patients were divided into group A(35%≤LVEF<50%)and group B (LVEF<35%). The differences of measurements were compared between two groups. The correlation between global myocardial strain in all direc?tions and left ventricular ejection fraction was analyzed. Results The GLS,GRS,GCS and GAS were significantly higher in group A than those in group B(P<0.01). The GLS,GRS,GCS and GAS were correlated with LVEF in group A(r=-0.871,0.610,-0.423,-0.797;P<0.05).The GCS,GRS and GAS were correlated with LVEF in group B(r=-0.517,0.368,-0.438;P<0.05). There was no significant correlation between GLS and LVEF in group B. Conclusion 3D?STE technology can be applied to evaluate the change of the myocardial movement. GLS is a promis?ing marker of the prognosis in patients with DCM.

Objective To develop the methods for identification and determination of scutellarin in compound Granule Tetrandra .Methods A TLC method was used to identify quality for scutellarin ,and a HPLC method was used to determine the content of scutellarin .Results The spot was clear in TLC identification without interference of negative control .The sam-ple size of scutellarin had a good linear relationship with peak area r=0 .9996(n=5)when ranged from 22 .4~156 .8 μg/ml , with the average recovery rate 97 .79% ( RSD=0 .32% ,n=6) .Conclusion The method is simple ,with good specificity and reproducibility ,which can be used as the quality control for this preparation .

Objective:To investigate the clinical phenotypes, therapy and genetic features of aldehyde dehydrogenase 7 family member A1 (ALDH7A1) gene mutations in five cases of pyridoxine dependent epilepsy (PDE) with diagnosis confirmed by next generation sequencing.Methods:Retrospective analysis was carried out on clinical data of five cases of PDE children with early epilepsy onset who were treated in the Department of Neurology of Children′s Hospital Affiliated to Zhengzhou University from February 2018 to November 2019. Next generation sequencing approach was used for genetic sequencing of proband ALDH7A1 gene and the first generation Sanger was used for validation of family members. And the characteristics of gene mutations were analyzed.Results:Among the five children diagnosed with PDE, the male to female ratio was 4 ∶ 1 and ages at clinic visit ranged from two months to 10 months old. In clinical phenotypes, all five cases experienced onset in neonatal period, with repeated seizures, manifested as myoclonus, spasms or focal paroxysm. The administration of antiepileptic drugs performed poorly in seizure control while long term oral intake of large dose pyridoxine showed better efficacy. All the five cases of children came from compound heterozygous mutations of father and mother, i.e. slicing homozygous mutation c.247-2(IVS2)A>T, missense mutation c.584A>G (p.N195S) and nonsense mutation c.1003C>T(p.R335 *), missense mutation c.1553G>C(p.R518T) and c.1547A>G(p.Y516C), missense mutation c.1547A>G(p.Y516C) and frameshift mutation c.1566_1568delTAC, missense mutation c.1061A>G(p.Y354C) and nonsense mutation c.841C>T(p.Q281X, 259), among which c.247-2(IVS2)A>T was novel splicing site mutation not reported before. Conclusions:PDE is induced by ALDH7A gene mutation. Early clinical manifestations are mostly onset of refractory epilepsy in neonatal period. Antiepileptic drugs perform poorly in terms of efficacy while pyridoxine can control seizure effectively. Gene analysis should be conducted on such patients for confirmed diagnosis.

Objective To study the effects of heat shock treatment of rat bone marrow mesenehymal stem cells(MSCs),the apoptosis ratio of treated-cells under low serum condition and the treated-cells transplantation on left ventricular function in rats with myocardiaIinfarction.Methods MSC8 were heat-treated under 42℃for 30 min,then the heat shock protein-70(HSP-70)was detected bv Western blot.The apoptosis ratio of heat-treated MSCs under low serum condition was tested by Annexin kit.The treated-MSCs labeled with Dil were transplanted into infarcted myocardium and 8 weeks later,the cardiac function of rats in each group was evaluated by echocardiography and cardiac catheterization.Results The immunophenotype of heat-treated MSCs did not vary,Western blot confirmed a higher level expression of HSP-70 in the treated-MSCs group as compared with that in the control group.The early apoptosis ratio was lower in treated-MSCs measured by flow cytometry with annexin staining than that of MSCs when cultured with low serum medium.After 8 weeks,LVEF,LVSP,+dp/dtmax,and-dp/dtmax were significantly higher,and the LVEDP was significantly lowar in heat-treated MSCs transplantation group than that in the control group.Conclusions Heat shock pretreatment of MSCs enhances the tolerance of MSCs to low serum medium,whereas does not lcad to the change of the cell immunophenotype.Transplantation of heattreated MSCs might improve the cardiac function in a rat myocardialinfarction model.

Objective:To explore the functional connections of the whole brain and the two hemispheres in patients with obsessive-compulsive disorder (OCD).Methods:Twenty-six patients with obsessive-compulsive disorder(patients group) and thirty-seven healthy controls matched in gender, age and education(control group) were enrolled.All the participants accepted the resting-state functional magnetic resonance (rs-fMRI) scan.Based on DPABI and REST software, degree centrality (DC) and voxel - mirrored homotopic connectivity (VMHC) approaches were used to explore the pattern of functional connection in OCD.Results:Compared with the control group, the DC values in the right posterior cerebellar lobe(MNI: x, y, z=45, -87, -12), left precentral gyrus(MNI: x, y, z=-54, 9, 39), left inferior parietal lobule(MNI: x, y, z=-48, -51, 42), right anterior cingulate cortex(MNI: x, y, z=3, 18, 48) were significantly higher( t values were 5.75, 5.26, 5.28 and 5.16, respectively), and the DC values in the left inferior frontal gyrus(MNI: x, y, z=-36, 9, 30) were significantly lower( t value was -6.65) in patients group.The VMHC values in bilateral posterior cerebellar lobe(MNI: x, y, z=±51, -69, -33), bilateral inferior parietal lobule(MNI: x, y, z=±48, -51, 54), bilateral anterior cingulate cortex(MNI: x, y, z=±3, 21, 45)in patients group were significantly higher that those in control group( t values were 5.19, 5.19, 5.02, 5.02, 5.15 and 5.15, respectively). The DC and VMHC values in patients group were not significantly correlated with clinical symptoms(-0.23< r<0.19, P>0.05). Conclusion:OCD patients have abnormal connections between key brain network nodes and relevant brain regions, and functional connections have increased among multiple cerebral hemispheres.

Objective:To investigate the clinical phenotype and genotypic characteristics of Legius syndrome.Methods:The clinical data of a child with precocious puberty and scattered café-au-lait macules admitted to Department of Neurology of the Children′s Hospital Affiliated to Zhengzhou University in July 2021 were retrospectively analyzed. Trio-whole exome sequencing (trio-WES) was used for genetic analysis to confirm the molecular diagnosis of the family. The relevant literature was reviewed to summarize the clinical characteristics of the disease.Results:The proband was a 10-year and 9-month-old girl, presenting with more than 5 café-au-lait macules with diameter>5 mm on the face and trunk, freckles in the axillary, without Lisch tubercles of iris and tumor signs of neurofibromatosis type 1, diagnosed as central precocious puberty at the age of 8. trio-WES results of the family revealed a spontaneous heterozygous nonsense mutation c.751(exon7) C>T in SPRED1 gene, causing a nonsense mutation in the amino acid sequence p.Arg251Ter (p. Ter251 *). Literature review showed a total of 88 pathogenic mutations were reported in SPRED1 gene, including frameshift mutations (41/88), nonsense mutations (31/88), splice mutations (7/88), missense mutations (6/88), and others (3/88), and no mutational hotspots were found. Clinical phenotype was as follows:>5 café-au-lait macules accounted for 92.8% (168/181), armpit and inguinal freckles 43.5% (73/168), macrocephaly 21.4% (31/145), learning disability 18.0% (30/166), psychomotor retardation 13.8% (22/159), lipoma (adult) 13.7% (21/153), Noonan facial sign 12.1% (21/173), and tumor phenotype of neurofibromatosis type 1 was not reported. Conclusions:The central precocious puberty phenotype of Legius syndrome was not reported in China. The clinical phenotype of Legius syndrome was mild, with a large variation, but without neurofibromatosis type 1 tumor phenotype. Genetic testing can be beneficial for early diagnosis of Legius syndrome.