AIM To establish a rapid quantitative analysis model for saponins in Paris polyphylla var.yunnanensis(PPY)by near infrared spectroscopy.METHODS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ and there total content in PPY were determined by HPLC,while spectral data within the range of 10 000 to 4 000 cm-1 were collected.A quantitative analysis model was established by combining these data with partial least squares regression(PLSR).Multivariate scatter correction(MSC)and vector normalization(SNV)were applied prior to further preprocessing the spectra with original,first-order derivative(1stD),or second-order derivative(2ndD)treatments.Lastly,the model was optimized through non-smoothing(NS),Norris Derivative filtering(Nd),and Savitzky-Golay filtering(S-G)method.Model stability was evaluated based on correlation coefficients and variance.The predicted contents of each saponin component in the validation set samples were calculated.RESULTS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ were 0.42-17.98,0.46-10.44,0.23-3.86 mg/g,respectively.The total content ranged from 2.91 to 22.1 mg/g.The optimal parameters of three saponins were achieved when selecting the MSC+2ndD+S-G pretreatment method.The corresponding ratio of line segment length to segment gap was 13∶5,15∶5,11∶5,with correlation coefficients of 0.982,0.930,0.958,respectively.The root mean square errors of calibration(RMSEC)were 0.702,0.797,0.238,and the root mean square errors of prediction(RMSEP)were 1.120,0.835,0.304,respectively.The optimal parameters for the total content were obtained when selecting the MSC+2ndD+NS pretreatment method,with a correlation coefficient of 0.970,a RMSEC of 1.090,and a RMSEP of 1.740.CONCLUSION This accurate and rapid method can be used for detection of saponin contents in P.Polyphylla.

AIM To establish a rapid quantitative analysis model for saponins in Paris polyphylla var.yunnanensis(PPY)by near infrared spectroscopy.METHODS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ and there total content in PPY were determined by HPLC,while spectral data within the range of 10 000 to 4 000 cm-1 were collected.A quantitative analysis model was established by combining these data with partial least squares regression(PLSR).Multivariate scatter correction(MSC)and vector normalization(SNV)were applied prior to further preprocessing the spectra with original,first-order derivative(1stD),or second-order derivative(2ndD)treatments.Lastly,the model was optimized through non-smoothing(NS),Norris Derivative filtering(Nd),and Savitzky-Golay filtering(S-G)method.Model stability was evaluated based on correlation coefficients and variance.The predicted contents of each saponin component in the validation set samples were calculated.RESULTS The contents of polyphyllins Ⅰ,Ⅱ,Ⅶ were 0.42-17.98,0.46-10.44,0.23-3.86 mg/g,respectively.The total content ranged from 2.91 to 22.1 mg/g.The optimal parameters of three saponins were achieved when selecting the MSC+2ndD+S-G pretreatment method.The corresponding ratio of line segment length to segment gap was 13∶5,15∶5,11∶5,with correlation coefficients of 0.982,0.930,0.958,respectively.The root mean square errors of calibration(RMSEC)were 0.702,0.797,0.238,and the root mean square errors of prediction(RMSEP)were 1.120,0.835,0.304,respectively.The optimal parameters for the total content were obtained when selecting the MSC+2ndD+NS pretreatment method,with a correlation coefficient of 0.970,a RMSEC of 1.090,and a RMSEP of 1.740.CONCLUSION This accurate and rapid method can be used for detection of saponin contents in P.Polyphylla.

Objective:To prepare 68Ga-2-(4, 7-bis(carboxymethyl)-1, 4, 7-triazonan-1-yl)pentanedioic acid (NODAGA)-YHWYGYTPQNVI (GE11) and evaluate its feasibility of PET imaging for pancreatic cancer. Methods:GE11 peptide was conjugated with NODAGA and then labeled with 68Ga. The labeling yield, radiochemical purity, hydrophilicity, stability and specificity in vitro were determined. Human pancreatic cancer BxPC3 nude mice models ( n=9) were established. MicroPET imaging was then obtained after 30 and 90 min, and mice were sacrificed at 90 min to acquire the radioactivity distribution of main organs and tumors. Pair t test was used to analyze the data. Results:The labeling yield was (73.5±5.4)% and radiochemical purity was more than 98%. After incubation 120 min in mouse serum at 37 ℃, radiochemical purity was more than 92%. The uptake was specific in BxPC3 cell lines. MicroPET images showed that 68Ga-NODAGA-GE11 could accumulate quickly in tumor. Value of tumor uptake was significantly higher than that of normal pancreas at 90 min ((1.38±0.25) vs (0.49±0.07) %ID/g; t=12.67, P<0.05), and the radio-uptake of blood, muscle and bone was lower than that of tumor. Conclusions:68Ga-NODAGA-GE11 is easy to be prepared with high radiochemical purity and good stability, and can specifically target BxPC3 xenograft tumor. However, due to the high uptake in the kidneys and liver, the value of 68Ga-NODAGA-GE11 in PET imaging for pancreatic tumor needs further study.

Objective:To study the correlation between changes of cerebral striatal dopamine D 2 receptors non-displaceable binding potential (BP ND), functional connectivity (FC) and clinical symptoms in patients with first-episode major depressive disorder (MDD), by 11C-Raclopride PET/CT and resting state fMRI (rs-fMRI). Methods:Thirty-eight first-episode depression patients (MDD group) and forty healthy volunteers (control group) matched with age, gender and years of education were selected. All subjects were scored with Hamilton depression scale (24 versions) before enrollment.All the subjects underwent cerebral 11C-Raclopride PET/CT and rs-fMRI in resting state. MIAKAT and DPARSF were used to analyze BP ND of cerebral striatal dopamine D 2 receptors and FC of striatum and the whole brain in subjects, respectively. Changes of striatal dopamine D 2 receptors BP ND and striatum and the whole brain FC of MDD were analyzed, and correlations among BP ND, FC and Hamilton depression rating scale were calculated by Rest 1.8 and SPSS 20.0. Results:Compared with the control group, BP ND of bilateral caudate nucleus and putamen dopamine D 2 receptors in the MDD group were decreased(left caudate nucleus: 1.16±0.37 vs 1.48±0.39, right caudate nucleus: 1.21±0.31 vs 1.62±0.48, left putamen: 1.73±0.47 vs 2.21±0.66, right putamen: 1.79±0.46 vs 2.17±0.65, t=3.66, -4.42, -3.68, -2.91, all P<0.001). Besides, FC of left caudate nucleus and left medial prefrontal lobes(4.38±1.31, 2.35±0.48), left caudate nucleus and left middle frontal gyrus(3.36±1.11, 1.64±0.56), left caudate nucleus and left superior frontal gyrus(3.14±0.78, 1.64±0.53), left putamen and left medial prefrontal lobes(4.10±1.42, 2.42±0.64, t=6.82, P<0.05), right caudate nucleus and right medial prefrontal lobes (4.32±1.30, 2.33±0.63, t=8.51, P<0.05), right putamen and right medial prefrontal lobes(3.77±1.25, 2.31±0.63, t=6.49, P<0.05)in the MDD group were increased.FC of left putamen and left anterior cingulate(1.60±0.55, 2.68±0.84, t=-6.76, P<0.05), right caudate nucleus and right amygdala (1.67±0.57, 3.46±0.64, t=-8.27, P<0.05) in the MDD group were decreased. Furthermore, there were significant negative correlations between D 2 receptors BP ND of bilateral striatum and FC of the same lateral striatum and medial prefrontal lobes ( r=-0.66, -0.50, -0.67, -0.47, all P<0.05). In MDD group, FC in left caudate nucleus and left medial prefrontal lobe were positively correlated with total score of Hamilton depression scale and anxiety somatization( r=0.55, 0.68, P<0.001). FC in left putamen and left medial prefrontal cortex were positively correlated with cognitive impairment and retardation ( r=0.37, 0.40, P=0.021, 0.001). FC of right caudate nucleus and right medial prefrontal lobe were positively correlated with Hamilton depression scale total score and anxiety somatization ( r=0.52, 0.67, all P<0.001). FC in right putamen and right medial prefrontal cortex was positively correlated with cognitive impairment ( r=0.50, P=0.002). Conclusion:The abnormal BP ND of cerebral striatal dopamine D 2 receptor of patients with first-episode depression is related to the abnormal activity of dopamine reward circuit related neurons in patients with MDD, which was related to clinical symptoms of depression. It may be involved in the pathogenesis of depression.

Objective To study the imaging features of 18F-fluorodeoxyglucose(FDG) PET/CT in patients with autoimmune encephalitis (AE) and evaluate the value of PET/CT on early diagnosis of AE.Methods Sixteen patients with AE (11 males,5 females,age:11-68 years) between March 2012 and December 2017 were included.Patients had positive antibodies in cerebrospinal fluid or (and) serum without immunity therapy.The imaging (18F-FDG PET/CT,MRI) and clinical data were analyzed.Results Nine patients suffered from anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis and other 7 patients had limbic encephalitis (LE),which including 2 cases of anti-leucine-rich glioma inactivated 1 (LGI1) encephalitis,3 cases of anti-γ-aminobutyric acid-B receptor(GABAsR) encephalitis,1 case of anti-Hu encephalitis and 1 case of anti-Yo encephalitis.Fifteen patients showed scattered hypermetabolism or hypometabolism in the brain on PET/CT imaging,and the positive rate was 15/16.Among those patients with anti-NMDAR encephalitis,hypermetabolism in frontotemporal parietal lobes and hypometabolism in occipital lobe were shown;hypermetabolism in limbic systems including temporal lobe and hippocampus were shown in LE.No abnormal CT density was found at the same phase.Slightly higher signals on T2,fluid-attenuated inversion recovery (FLAIR) and diffusion weighted imaging (DWI) were detected in some patients,and the positive rate was 7/16.Conclusions Patients with AE of different types have different characteristics on 18F-FDG PET/CT.18F-FDG PET/CT has high positive rate for early diagnosis of AE.

Objective To explore the changes of dopamine D2 receptor in dopamine pathway in in-somnia patients and discuss its clinical significance. Methods From January 2016 to December 2016, 15 patients with insomnia (1 male, 14 females, age:(44.3±8.6) years) and 15 gender-/age-matched-healthy volunteers (control group;3 males, 12 females, age:(40.5±9.0) years) were included to undergo resting brain 11C-Raclopride PET/CT imaging. The D2 receptor binding potential (BPND) of the dopamine pathway was calculated by molecular imaging and kinetic analysis toolbox ( MIAKAT) software. The BP ND , Hamilton depression scale ( HAMD) , transient and graphics memory scale results were compared with two-sample t test and Mann-Whitney u test between the two groups. Pearson correlation analysis was used to evaluate the correlation between BPND(nucleus accumbens, caudate nucleus, putamen) and Pittsburgh sleep quality in-dex ( PSQI) , HAMD, course of disease, transient memory and graphical memory scale scores in the patient group. Results The BP ND in bilateral putamen, nucleus accumbens and left caudate nucleus of patients was lower than that of controls( left putamen:z=-2.717, right putamen:z=-2.883, both P<0.01;left nu-cleus accumbens:t=-2.269, right nucleus accumbens:t=-2.410, both P<0.05;left caudate nucleus:t=-2.632,P<0. 05), but the BPND level of right caudate nucleus was not significantly different(z=-0.850, P>0.05) . The scores of HAMD in the patient group were higher than those in control group ( t=10. 273, P<0. 01), while the scores of instantaneous memory (t=-4.888, P<0.01) and graphical memory scale (t=-2.624, P<0.05) were lower. There were significant negative correlations between the BP ND of bilateral nucleus ac-cumbens, caudate nucleus and putamen and the course of insomnia in the patient group ( r range:-0.761 to-0.682, all P<0.01) . Conclusion Patients with insomnia have abnormal neurotransmitter system of dopa-mine D2 and it may play a role in the pathogenesis of insomnia.

Objective To explore the correlation between changes of striatal dopamine D2receptors non-displaceable binding potential (BPND) in 11C-Raclopride PET-CT and brain regional homogeneity (ReHo) derived from resting state functional MRI (fMRI) in patients with first-episode major depressive disorder (MDD) . Methods Patients with first-episode MDD and age and sex matched healthy volunteers accepted brain 11C-Raclopride PET-CT and resting state fMRI. MIAKAT based on MATLAB and Rest 1.8 were used to calculate BPNDof brain dopamine D2receptors and brain ReHo, respectively. Changes of striatal dopamine D2receptors BPNDand brain ReHo values were analyzed by paired-sample t test and two independent sample t-test, and Pearson correlation analysis was used to analyze the correlation between BPNDand ReHo. Results Twenty patients with first-episode MDD were enrolled as MDD group, and 20 healthy volunteers as the control group. The two groups were all right-handed, and there were no statistical differences for age (t =1.33,P=0.19)and gender(χ2=0.10,P=0.75). Compared with the control group, the ReHo in MDD patients increased in the bilateral striatum (caudate nucleus and putamen), bilateral medial prefrontal lobes, and right thalamic (27 to 56 voxels, P＜0.05) and decreased in the left middle frontal gyrus, the left anterior cingulate, the left hippocampal and the right amygdala (21 to 35 voxels, P＜0.05). In addition, compared with the control group, BPNDof bilateral caudate nucleus and putamen dopamine D2receptors in the MDD group were decreased (t=－4.41 to －3.13, P＜0.05). Furthermore, there was a negative correlation between D2 receptors BPNDand ReHo of bilateral caudate nucleus and putamen (r=－0.81 to－0.62, P＜0.05). And there was a negative correlation between ReHo of the bilateral medial prefrontal lobes and BPNDof the same lateral caudate nucleus and putamen D2receptors in the MDD group (r=－0.86 to－0.52, P＜0.05). Besides, ReHo of the left middle frontal gyrus, right thalamic, left anterior cingulate, left hippocampal and right amygdala had no correlation with the D2receptors BPNDof the striatum in the MDD group (－0.27 to 0.39, P＞0.05). Conclusion There were abnormal levels of dopamine neurotransmitter in the cerebral striatum regions and abnormal brain activities in the brain region associated with dopamine reward circuit in the first-episode MDD patients, and there was a correlation between the two abnormalities.

Objective To observe non-displaceable binding potential (BPND) changes of striatal dopamine D2 receptors(SDDR) in patients with first-episode major depressive disorder (MDD) using 11C-Raclopride PET/CT,and to analyze the relationship between BPND and Hamilton rating scale for depression (HAM-D).Methods From December 2014 to December 2015,patients with first-episode MDD and age/gender-matched healthy controls underwent brain MRI and 11C-Raclopride PET/CT in this prospective study.BPND of bilateral SDDR was calculated by molecular imaging and kinetic analysis toolbox (MIAKAT).BPND changes of bilateral SDDR and their relationship with HAM-D score were analyzed.Paired t test,two-sample t test and Pearson correlation analysis were used.Results A total of 20 MDD patients (8 males,12 females,average age: (32.80±9.76) years) and 20 healthy controls (9 males,11 females,average age:(29.25±6.93) years) were enrolled in this study.The 11C-Raclopride uptake in brain tissue of the MDD group and control group were mainly distributed in bilateral striatum,and very few 11C-Raclopride was distributed in bilateral cerebral cortex and cerebellum.In MDD group,the BPND level of bilateral SDDR had no statistical differences(t values: 0.69,0.35,both P>0.05),and similar results were found in the control group(t values: 0.28,0.24,both P>0.05).Compared with the control group,however,the MDD group had lower BPND level of bilateral SDDR(t values: 3.13-4.41,all P<0.05).The BPND of bilateral caudate nucleus and/or putamen D2 receptors was correlated with HAM-D total score,anxiety/somatization factor score,cognitive impairment factor score,retardation factor score and sleep disturbance factor score(r values: from-0.688 to-0.453,all P<0.05).Conclusions The binding potential of SDDR in patients with first-episode MDD is declined,and the BPND level of SDDR is correlated with symptoms of depression.The abnormality of SDDR may be an important molecular mechanism of the abnormality of midbrain-striatal dopamine reward circuits in MDD patients.

Objective: To investigate the essential biochemical indices like 1 -hydroxylase and hypocalcaemia in the rats with severe acute pancreatitis and explore the correlation between them. Methods: A total of 120 SPF grade Wistar male rats which were in similar physiological status were selected and randomly divided into two groups: sham group (SO group) and severe acute pancreatitis group (SAP group). Then they were divided into 1 h, 3 h, 6 h, and 12 h subgroups according to the killing time. The severe acute pancreatitis model was established by retrograde injection of 5% sodium taurocholate. Serum calcium, serum creatinine, serum urea nitrogen and serum amylase were measured at different time. Serum 1, 25 dihydroxy vitamin D3 level was determined by enzyme linked immunosorbentassay. The expression of 1-hydroxylase protein in the kidney tissue was determined with Western blotting and immunohistochemistry to observe its location. The pathologic features of the kidney tissue section was observed under light microscope and submicroscopic structure of the proximal convoluted tubule epithelial cell was observed under transmission electron microscope. Results: Compared with the SO group, rats in the SAP group showed continuous pathological injury as time went by. There was significant increase in serum creatinine, serum urea nitrogen and serum amylase in SAP group compared with the SO group 1, 3, 6, 12 hours after the operation (P<0.05). There was significant decrease in serum calcium and 1, 25 dihydroxy vitamin D3 3, 6, 12 hours after the operation (P<0.05). It also showed that the expression of the 1-hydroxylase protein in kidney tissues was upregulated at 1 h, 3 h and decreased at 6 h, 12 h compared with the SO group. The serum calcium, 1, 25 dihydroxy vitamin D3 and the expression of the 1-hydroxylase protein in kidney tissues of the SAP group showed sustaining decrease. Western blotting showed positive correlation between the 1-hydroxylase expression and serum calcium at 3 h, 6 h and 12 h (r=0.976, P<0.001; r=0.948, P<0.001; r=0.742, P=0.001) and also positive correlation between the 1-hydroxylase expression and serum 1, 25 dihydroxy vitamin D3 at 1 h, 3 h, 6 h and 12 h (r=0.935, P<0.001; r=0.952, P<0.001; r=0.917, P<0.001; r=0.874, P<0.001). Conclusions: At the early stage of the kidney injury, the expression of 1-hydroxylase in the kidney tissue is reduced with the progress of the disease and the decrease in its activity has a correlation with the hypocalcaemia.

AIM: To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae). METHODS: The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking. RESULTS: Wikstroelide M potently inhibited different HIV-1 strains, including HIV-1IIIB, HIV-1A17, and HIV-19495, induced a cytopathic effect, with EC50 values ranging from 3.81 to 15.65 ng·mL⁻¹. Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL⁻¹. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with EC50 values ranging from 15.16 to 35.57 ng·mL⁻¹. Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells, with an EC50 value of 9.60 ng·mL⁻¹. The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75. CONCLUSION Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.
Anti-HIV Agents ; pharmacology ; therapeutic use ; Cell Line ; Daphne ; chemistry ; Diterpenes ; pharmacology ; HIV Infections ; drug therapy ; virology ; HIV Integrase ; metabolism ; HIV Integrase Inhibitors ; pharmacology ; therapeutic use ; HIV Reverse Transcriptase ; antagonists & inhibitors ; HIV-1 ; drug effects ; enzymology ; HIV-2 ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Virus Integration ; drug effects ; Virus Replication ; drug effects