This research use "3414" fertilizer effect experiments to handle zinc, boron and molybdenum trace element fertilizer, determined the dry matter accumulation and content of saikosaponion a and d, to investigate the different ratio of zinc, boron and molybdenum on yield and saikosaponin a, saikosaponin d contents of Bupleurum chinense. Found The suitable ratio of zinc, boron and molybdenum play an active role on dry matter accumulation and distribution, the treatment Zn2B2Mo3 is the best one to promote the dry matter accumulation and transfer to the underground part; in a certain range, only use zinc or molybdenum can promote the yield of B. chinense, the yield of treatment Zn2B2Mo1 is the highest one. According to the results of regression analysis: in accordance with Zn 48.45 g x hm(-2), B 355.05 g x hm(-2), Mo 86.40 g x hm(-2), can obtain the yield with 3313.05 kg x hm(-2); the treatment Zn2BMo2 is most effective to promote the total saikosaponin a and d accumulated, according to the results of regression analysis: in accordance with Zn 36.15 g x hm(-2), B 343.05 g x hm(-2), Mo 106.35 g x hm(-2), the content of total saikosaponin a and d can reach 1.23%. This research first discovered the suitable ratio of zinc, boron and molybdenum can promote the yield and saikosaponin a, saikosaponin d contents on B. chinense.
Boron ; metabolism ; Bupleurum ; metabolism ; Fertilizers ; Molybdenum ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Roots ; metabolism ; Saponins ; metabolism ; Trace Elements ; metabolism ; Zinc ; metabolism

In order to select high quality and suitable Bupleuri Radix varieties in Qingchuan, and establish a new comprehensive method to evaluation the quality of Bupleuri Radix, 12 characters of 14 samples were evaluated by DTOPSIS and grey related degree. The results showed that varieties No. 8 and No. 10 had high quality. DTOPSIS and grey related degree gave the uniformity result, and the biggest difference of value of Ci in DTOPSIS method was 46. 33% , but the biggest difference of the weighting correlation number( r (i)) in grey related degree was only 13.10%. The DTOPSIS combined with grey related degree can evaluate the quality of Bupleuri Radix comprehensively and objectively.
Bupleurum ; chemistry ; China ; Drugs, Chinese Herbal ; chemistry ; supply & distribution ; Quality Control ; Statistics as Topic ; methods

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) and determine its physical and chemical characters, recombinant NDPK-A producing E. coli was cultured in 80L fermentor under high cell density culture (HCDC) conditions. The harvested cells were treated with high pressure to break the cell up, tangential-flow microfiltration to remove the bacteria debris and ultrafiltration to concentrate the filtered solution containing target protein. The crude NDPK-A was purified by ion exchange chromatography with DEAE Sepharose Fast Flow, affinity chromatography with Cibarcron Blue 3GA Sepharose CL-4B and gel filtration with Sephadex G-100. The purity of rhNDPK-A was analyzed with SDS-PAGE and RP-HPLC. The Enzymatic activity was determined with RP-HPLC. The molecular weight (MW) was measured with matrix assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS). The N-terminal residue was sequenced with Edman method. The apparent molecular weight of rhNDPK-A in solution was determined with multiangle laser light-scattering method (MALS). It was found that the purity of rhNDPK-A was 97.3% with SDS-PAGE method and 99.2% with RP-HPLC method. The specific enzymatic activity was (900 +/- 100) u/mg. The molecular weight was 17017, which was 132 less than the calculated value according to the amino acid sequence of NDPK-A. The sequencing result of rhNDPK-A revealed that its N-terminal residue was Ala, which was the second residue on N-terminal of native NDPK-A. The calculated MW of N-terminal deleted rhNDPK-A was 17017, exactly equal to the experimental value. The result of apparent MW determination revealed that rhNDPK-A formed homohexamer in solution with a MW of 102kD. These results suggested that rhNDPK-A possessed character identical to its native counterpart of assembling into hexamer. Confirming the identity of rhNDPK-A to its native counterpart provided a good foundation for drug development and mechanism study of NDPK-A.
Amino Acid Sequence ; Humans ; Molecular Sequence Data ; Molecular Weight ; NM23 Nucleoside Diphosphate Kinases ; chemistry ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; isolation & purification ; Scattering, Radiation ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Objective:To find out the difference between children with attention-deficit/hyperactivity disorder and normal control,and the difference between boys and girls with ADHD in resting-state.Methods:There were 38 children with ADHD in this study including 31 boys and 7 girls,meanwhile,42 normal controls whose age and gen der were matched with children with ADHD were involved too,including 29 boys and 13 girls.Total scores of ADHD rating scale (ADHD-RS) were used to measure the severity of symptom for both ADHD and normal control.Magnetic resonance imaging was used to collect the resting-state functional magnetic resonance imaging data,and the fractional amplitude of low frequency fluctuation (fALFF) was measured for each child.Analysis of covariance (ANCOVA) was used to find out the difference between the group and the sex.Results:Children with ADHD showed a higher fALFF in the right medial superior frontal gyrus and right supplementary motor area,and a lower fALFF in the left putamen,bilateral cerebellum and bilateral precuneus than controls (all corrected P ＜0.05).ADHD boys showed a lower fALFF in the left cerebellum and a higher fALFF in the bilateral medial superior frontal gyrus than boys without ADHD (all corrected P ＜0.05).ADHD girls showed a higher fALFF in the bilateral cerebellum and in the thalamus than girls without ADHD,and a lower fALFF in the insula than girls without ADHD (all corrected P ＜ 0.05).There was a negative relationship between the fALFF of the left cerebellum of ADHD girls and the scores of inattention symptoms.Conclusion:The fALFF difference exist between ADHD and normal controls,and between boys and girls with ADHD.

Objective:To investigate the relations of serum hypoxia inducible factor 2α (HIF-2α) and miR-21 expressions with cerebral vasospasm in patients with aneurysmal subarachnoid hemorrhage (aSAH) after interventional embolization.Methods:One hundred and seventy-four patients with aSAH underwent interventional embolization in our hospital from October 2017 to June 2019 were prospectively selected. DSA examination was performed 4 d after surgery, and severity of cerebral vasospasm was evaluated. Enzyme-linked immunosorbent assay was used to detect the level of serum HIF-2α and reverse transcription-PCR was employed to detect the serum miR-21 expression before and 3 and 7 d after interventional embolization. The clinical data and changes of serum HIF-2α and miR-21 expressions in patients with different degrees of cerebral vasospasm were compared. Correlation analysis was performed to analyze the relation of HIF-2α expression with miR-21 expression 3 and 7 d after interventional embolization. Receiver operating characteristics curve was used to analyze the diagnostic values of serum HIF-2α and miR-21 levels in cerebral vasospasm 3 d after interventional embolization.Results:There were 100 patients without vasospasm, and 20, 38 and 16 patients with mild, moderate and severe cerebral vasospasm, respectively. The serum levels of HIF-2α and miR-21 in patients with mild, moderate and severe cerebral vasospasm increased successively 3 and 7 d after interventional embolization, with significant differences ( P<0.05). Positive correlation was noted between expressions of HIF-2α and miR-21 in serum 3 and 7 d after interventional embolization ( P<0.05). Area under the curve (AUC) of HIF-2α in diagnosis of cerebral vasospasm was 0.748 ( 95%CI: 0.615-0.883, P=0.000) 3 d after interventional embolization. AUC of serum miR-21 level in diagnosis of cerebral vasospasm was 0.715 ( 95%CI: 0.590-0.842, P=0.000). AUC of serum HIF-2α combined with miR-21 in diagnosis of cerebral vasospasm was 0.893 ( 95%CI: 0.792-0.985, P=0.000). When diagnostic critical points of HIF-2α and miR-21 were 82.75 pg/mL and 1.15, the sensitivity, accuracy and negative predictive value of HIF-2α combined with miR-21 in the diagnosis of cerebral vasospasm were higher than those of HIF-2α or miR-21 alone. Conclusion:The expressions of serum HIF-2α and miR-21 in patients with aSAH after interventional embolization can effectively predict the occurrence of cerebral vasospasm, and may be involved in the occurrence and development of cerebral vasospasm.

To express and secrete native HBscFv (anti-HBsAg single-chain Fv) in P. pastoris, HBscFv was amplified from plasmid pGEM-HBscFv, and then sub-cloned into expression vector pPICZalphaA. The resulting plasmid pPIC-HBscFv was linearized and transformed into P. pastoris GS115. The recombinant Pichia strains, identified by direct PCR and Zeocin-resistant screening of Pichia transformants, were cultured and induced with methanol. It was found that recombinant HBscFv, lead by alpha-factor, could be secreted into the culture supernatant to a level of 80mg/L. The bioactivity of Pichia produced HBscFv was confirmed by indirect ELISA, which also suggested that the bioactivity of HBscFv in the culture supernatant reached its peak in 72h and decreased in the late-stage of the induction. PAS staining suggests that HBscFv produced by yeast is poorly glycosylated or none-glycosylated protein.
Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Hepatitis B Surface Antigens ; immunology ; Humans ; Pichia ; genetics ; metabolism ; Polymerase Chain Reaction ; Single-Chain Antibodies ; genetics ; immunology ; metabolism

BACKGROUNDNeuroimaging studies have found that functional changes exist in patients with Parkinson's disease (PD). However, the majority of functional magnetic resonance imaging (fMRI) studies in patients with PD are task-related and cross-sectional. This study investigated the functional changes observed in patients with PD, at both baseline and after 2 years, using resting-state fMRI. It further investigated the relationship between whole-brain spontaneous neural activity of patients with PD and their clinical characteristics.

METHODSSeventeen patients with PD underwent an MRI procedure at both baseline and after 2 years using resting-state fMRI that was derived from the same 3T MRI. In addition, 20 age- and sex-matched, healthy controls were examined using resting-state fMRI. The fractional amplitude of low-frequency fluctuation (fALFF) approach was used to analyze the fMRI data. Nonlinear registration was used to model within-subject changes over the scanning interval, as well as changes between the patients with PD and the healthy controls. A correlative analysis between the fALFF values and clinical characteristics was performed in the regions showing fALFF differences.

RESULTSCompared to the control subjects, the patients with PD showed increased fALFF values in the left inferior temporal gyrus, right inferior parietal lobule (IPL) and right middle frontal gyrus. Compared to the baseline in the 2 years follow-up, the patients with PD presented with increased fALFF values in the right middle temporal gyrus and right middle occipital gyrus while also having decreased fALFF values in the right cerebellum, right thalamus, right striatum, left superior parietal lobule, left IPL, left precentral gyrus, and left postcentral gyrus (P < 0.01, after correction with AlphaSim). In addition, the fALFF values in the right cerebellum were positively correlated with the Unified PD Rating Scale (UPDRS) motor scores (r = 0.51, P < 0.05, uncorrected) and the change in the UPDRS motor score (r = 0.61, P < 0.05, uncorrected).

CONCLUSIONSThe baseline and longitudinal changes of the fALFF values in our study suggest that dysfunction in the brain may affect the regions related to cortico-striato-pallido-thalamic loops and cerebello-thalamo-cortical loops as the disease progresses and that alterations to the spontaneous neural activity of the cerebellum may also play an important role in the disease's progression in patients with PD.

Adult ; Aged ; Brain ; pathology ; Cross-Sectional Studies ; Female ; Humans ; Longitudinal Studies ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Parkinson Disease ; diagnosis

Objective: Attention-deficit/hyperactivity disorder (ADHD) is a common neurodevelopmental disorder in children and adolescents. The present study investigated the cortical morphology features and their relationship with working memory (WM). Methods: In the present study, a total of 36 medication naïve children with ADHD (aged from 8 to 15 years) and 36 age- and gendermatched healthy control (HC) children were included. The digit span test was used to evaluate WM. The magnetic resonance imaging (MRI) was used to examine the characteristics of cortical morphology. Firstly, we compared the cortical morphology features between two groups to identify the potential structural alterations of cortical volume, surface, thickness, and curvature in children with ADHD. Then, the correlation between the brain structural abnormalities and WM was further explored in children with ADHD. Results: Compared with the HC children, the children with ADHD showed reduced cortical volumes in the left lateral superior temporal gyrus (STG) (p=6.67×10-6) and left anterior cingulate cortex (ACC) (p=3.88×10-4). In addition, the cortical volume of left lateral STG was positively correlated with WM (r=0.36, p=0.029). Conclusion Though preliminary, these findings suggest that the reduced cortical volumes of left lateral STG may contribute to the pathogenesis of ADHD and correlate with WM in children with ADHD.

Objective: Attention-deficit/hyperactivity disorder (ADHD) is a common neurodevelopmental disorder in children and adolescents. The present study investigated the cortical morphology features and their relationship with working memory (WM). Methods: In the present study, a total of 36 medication naïve children with ADHD (aged from 8 to 15 years) and 36 age- and gendermatched healthy control (HC) children were included. The digit span test was used to evaluate WM. The magnetic resonance imaging (MRI) was used to examine the characteristics of cortical morphology. Firstly, we compared the cortical morphology features between two groups to identify the potential structural alterations of cortical volume, surface, thickness, and curvature in children with ADHD. Then, the correlation between the brain structural abnormalities and WM was further explored in children with ADHD. Results: Compared with the HC children, the children with ADHD showed reduced cortical volumes in the left lateral superior temporal gyrus (STG) (p=6.67×10-6) and left anterior cingulate cortex (ACC) (p=3.88×10-4). In addition, the cortical volume of left lateral STG was positively correlated with WM (r=0.36, p=0.029). Conclusion Though preliminary, these findings suggest that the reduced cortical volumes of left lateral STG may contribute to the pathogenesis of ADHD and correlate with WM in children with ADHD.

To explore the anti-HSV-1 effect of silencing gD gene expression by RNA interference, five 21-nucleotide duplex small interfering RNAs(siRNAs) targeting the HSV1 gD sequence were designed and the gD-EGFP fusion gene expression vector was constructed, then co-transfected into Vero cell, and screened the effective siRNA through analyzing the intensity of the EGFP fluorescence. Finally, the anti-HSV1 effect was confirmed by plaque reduction assay, real-time PCR and daughter virus titration of HSV1 infected Vero cells transfected with siRNAs. The study demonstrated that siRNAs could effectively and specifically inhibit gD gene expression in HSV1-infected cells, but only had a little effect on HSV1 infection, so taking gD as the target of siRNA against HSV1 needs further study.
Animals ; Cercopithecus aethiops ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Herpesvirus 1, Human ; genetics ; Humans ; Polymerase Chain Reaction ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; Vero Cells ; Viral Envelope Proteins ; genetics ; metabolism