Screening of quantitative preparation quality markers of Zhibao Sanbian Wan / 药学学报

On the basis of the qualitative preparation quality markers of Zhibao Sanbian Wan (ZBSBW), we screened out the quantitative markers and evaluated the content consistency of ZBSBW. A method capable of simultaneously determining 34 compounds in ZBSBW was established based on HPLC-MS/MS, and 16 batches of ZBSBW were simultaneously analyzed by this method. Furthermore, we explored a general strategy for analyzing the component migration in preparation, plasma, brain tissue and cerebrospinal fluid. The methodological investigation was confirmed by linear range, recovery (85.10%-105.07%), precision (RSD: 1.37%-4.58%), stability, and repeatability (3.00%-12.45%), the established method was suitable for the detection and quantification of the compounds in ZBSBW. The contents of compounds in ZBSBW were all lower than 1 mg·g-1, and the contents and daily dose of nystose were the highest, followed by echinacoside, paeoniflorin, osthole and paeonol. The results of systematic clustering showed that the contents were consistent for ordinary preparations of ZBSBW. The principal component analysis showed that the components of berberine, ginsenoside Re, ginsenoside Rg1, pinoresinol diglucoside and tenuifolin had large variation, which contributed significantly to the grouping. The contents of echinacoside, verbascoside, polygalaxanthone Ⅲ, β-ecdysterone, osthole, alisol B 23-acetate, liquiritin and glycyrrhizic acid were stable from batch to batch. The animal experiment results showed that osthole, paeonol and liquiritin in ZBSBW could be absorbed into the blood and enter the brain tissue by passing through the blood-brain barrier. All animal studies were reviewed and approved by the Institutional Animal Care and Use Committee at Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (No. 2020B071). The above compounds contributed the quantitative preparation quality markers of ZBSBW. In conclusion, the HPLC-MS/MS method established in this study was sensitive, accurate and rapid, and could be used for simultaneous quantification of 34 compounds and content consistency evaluation of multiple batches of preparations in ZBSBW. The result provided a methodological basis for the screening of quantitative preparation quality markers and material basis research of ZBSBW.

Assessment of conventional magnetic resonance morphological measurements in diagnosis and differential diagnosis of progressive supranuclear paralysis / 中华老年医学杂志

Objective:To study the features as well as the diagnosis and differential diagnosis values by conventional MRI morphometrics in different clinical subtypes of progressive supranuclear palsy(PSP).Methods:Forty five patients with PSP were included, comprising three PSP subtypes: 15 cases of Richardson's syndrome(PSP-RS), 15 cases of Parkinson's syndrome(PSP-P)and 15 cases of progressive frozen gait(PSP-PFG). In addition, three control groups were established: 15 cases of multiple system atrophy-Parkinson's syndrome(MSA-P), 30 cases of primary Parkinson's disease(PD)and 40 healthy controls(HC). Midbrain area-to-Pons area ratio(M/P), Magnetic Resonance Parkinsonism Index(MRPI, MRPI2.0), width ratio of middle cerebellar peduncle to superior cerebellar peduncle(MCP/SCP), Midbrain-to-Pons ratio(MTPR), Angle of cerebral peduncle(A cp), third ventricle width/frontal horns width ratio(V 3rd/FH), and Humming bird sign rating scale(HBS-RS)scores were calculated.Diagnostic sensitivity and specificity were performed by ROC curve to assess the accuracy of these imaging indicators in the diagnosis and differential diagnosis of PSP and its subtypes. Results:The MRPI, MRPI2.0, MCP/SCP and HBS-RS scores were significantly higher in PSP group than in other control groups( H=69.351, 66.776, 33.926 and 84.694, all P<0.05), while M/P and MTPR were significantly lower in PSP group than in other control groups(H=60.101 and 77.276, all P<0.05). PSP group also had higher V 3rd/FH compared with PD or HC group( F=17.168, P<0.05), but not with MSA-P group( Z=-1.602, P>0.05). The above differences also existed between each PSP subgroup and control groups.Among PSP subgroups, PSP-PFG subgroup had a larger A cp than did PSP-RS( Z=-2.510, P<0.05), and had higher HBS-RS score than did PSP-P group( Z=-2.380, P<0.05). No significant differences in other MRI morphometric indexes were identified among PSP subtypes.The M/P, MRPI, MTPR, MRPI2.0, HBS-RS score showed good accuracy in diagnosing PSP and its each subgroup, with HBS-RS score being the most accurate indicator, when the cutoff value was 2, the AUC values were all higher than 0.99, and the sensitivity and specificity were all above 90%.PSP and its subtypes were best distinguished from MSA-P by MRPI, when the cutoff value was 9.94, the AUC values were all higher than 0.90, with the sensitivity of 100% and specificity of 86.67%.PSP and its subtypes were best distinguished from PD by MTPR, AUC values were all above 0.95, with slightly different cutoff values.Almost all the morphological measurement parameters failed to show significant sensitivity and specificity in discriminating subtypes of PSP.The sensitivity and specificity of almost all MRI morphometry indicators in differentiating different subtypes of PSP are not high. Conclusions:MRI morphometrics have a high value both in the diagnosis of PSP and its subtypes, and also in specific application fields.MRI morphometrics have a limited value in discriminating PSP subtypes.

Effect of transcutaneous auricular vagus nerve stimulation on nocturnal autonomic nervous function in primary insomnia patients / 中国针灸

OBJECTIVE@#To observe the effect of transcutaneous auricular vagus nerve stimulation (taVNS) on the sleep quality and nocturnal heart rate variability (HRV) in patients with primary insomnia.@*METHODS@#Twenty-one patients with primary insomnia were included. Using SDZ-ⅡB electric acupuncture apparatus, Xin (CO15) and Shen (CO10) were stimulated with disperse-dense wave, 4 Hz/ 20 Hz in frequency, (0.2±30%) ms of pulse width and tolerable intensity. Electric stimulation was given once every morning and evening of a day, 30 min each time, for 4 weeks totally. Before and after treatment, the score of Pittsburgh sleep quality index (PSQI), objective sleep structure (total sleep time [TST], sleep latency [SL], wake after sleep onset [WASO], sleep efficiency [SE], the percentages of non-rapid eye movement period 1, 2, 3, and the percentage of rapid eye movement period to TST [N1%, N2%, N3%, REM%] ) and nocturnal HRV (high frequency [HF], low frequency [LF], the ratio of LF to HF [LF/HF], standard deviation for the normal RR intervals [SDNN], squared root of the mean sum of squares of differences between adjacent intervals RR [RMSSD], the percentage of adjacent RR intervals with differences larger than 50 ms in the entire recording [PNN50%], the mean of sinus RR intervals [NNMean] ) were compared in the patients separately.@*RESULTS@#After treatment, the score of each item and the total score of PSQI and SL were all reduced as compared with those before treatment (P<0.01, P<0.001); SE, N3%, LF, HF, LF/HF, SDNN, NNMean and RMSSD were all increased compared with those before treatment (P<0.001, P<0.01).@*CONCLUSION@#The taVNS improves the sleep quality and objective sleep structure in patients with primary insomnia, which is probably related to the regulation of autonomic nervous functions.

Three new ursane-type triterpenoids from Rosmarinus officinalis and their biological activities / 中国天然药物

Three new ursane-type triterpenoids, 3-oxours-12-en-20, 28-olide (1), 3β-hydroxyurs-12-en-20, 28-olide (2) and 3β-hydroxyurs-11, 13(18)-dien-20, 28-olide (3), were isolated from a potent anti-inflammatory and antibacterial fraction of the ethanolic extract of Rosmarinus officinalis. Their structures were elucidated by a combination of extensive 1D- and 2D-NMR experiments, MS data and comparisons with literature reports. Compounds 1-3 exhibited significantly inhibitory effects on nitric oxide production in lipopolysaccharide-activated mouse RAW264.7 macrophages, but no antibacterial activity was found at a concentration of 128 μg·mL-1.

Mechanism of Chuanxiong Rhizoma intervention on central sensitization of Panx1-Src-NMDAR-2B signaling pathway in neuropathic pain model rats / 中国中药杂志

Excitatory toxicity(ET) is an important factor of neuropathic pain(NPP) induced by central sensitization(CS), and the association of pannexin-1(Panx1)-Src-N-methyl-D-aspartate receptor subunit 2 B(NMDAR-2 B) is an important new pathway for ET to initiate CS. The present study confirmed whether the central analgesic effect of Chuanxiong Rhizoma extract(CRE) was achieved through the synchronous regulation of the brain and spinal pathways of Panx1-Src-NMDAR-2 B. In this study, dynamic and simulta-neo-us microdialysis of the brain and spinal cord in vivo combined with behavioristics, high performance liquid chromatography(HPLC)-fluorescence detection, microdialysis analysis(ISCUS~(flex)), ultrasensitive multifactorial electrochemiluminescence immunoassay, ELISA, and Western blot was employed to investigate the protein expression of NMDAR-2 B, Src, and Panx1, extracellular excitatory amino acids, cytokines, energy metabolites, and substance P in spinal dorsal horn(SDH) and anterior cingulate cortex(ACC) after CRE intervention with the rat model of spared sciatic nerve injury(SNI) as the experimental tool. Compared with the sham group, the SNI group exhibited diminished mechanical withdrawal threshold(MWT)(P<0.01), increased cold spray scores(P<0.01), glutamate(Glu), D-serine(D-Ser), and glycine(Gly) in extracellular fluids of ACC, and Glu, D-Ser, interleukin-1β(IL-1β), and lactic acid(Lac) in extracellular fluids of SDH(P<0.05), dwindled tumor necrosis factor(TNF-α)(P<0.05), and elevated protein levels of NMDAR-2 B, Src, and Panx1 in ACC(P<0.05). Compared with the SNI model rats, high-and medium-dose CRE(CRE-H/M) could potentiate the analgesic activity as revealed by the MWT test(P<0.05) and CRE-M enabled the decrease in cold spray scores(P<0.05). CRE-H/M could inhibit the levels of Glu, D-Ser and Gly in the extracellular fluids of ACC(P<0.05), and the levels of Glu in the extracellular fluids of SDH(P<0.05) in SNI rats. CRE-M significantly increased the levels of glucose(Gluc), Lac, interferon-gamma(IFN-γ), keratinocyte chemoattractant/human growth-regulated oncogenes(KC/GRO), and IL-4 in extracellular fluids of SDH in SNI rats(P<0.05). CRE-H/M/L could also inhibit the levels of NMDAR-2 B, Src and Panx1 in ACC and SDH in SNI rats(P<0.05). The central analgesic effect of CRE is presumedly related to the inhibited release of excitatory amino acid transmitters(Glu, D-Ser and Gly) in ACC and SDH of SNI rats, decreased protein expression of NMDAR-2 B, Src and Panx1 in the two regions, and the regulation of the Panx1-Src-NMDAR-2 B pathway in the spinal cord and brain. The above findings partially clarified the scientific basis of clinical analgesic effect of Chuanxiong Rhizoma.

Observation of acupoint application of gel plaster on insomnia / 中国针灸

OBJECTIVE@#To observe the effect of acupoint application of gel plaster on quality of sleep and life in patients with insomnia.@*METHODS@#A total of 63 patients with insomnia were randomized into a gel plaster group (32 cases, 1 case dropped off) and a placebo plaster group (31 cases). Acupoint application of gel plaster was applied at Yintang (GV 29) and Yongquan (KI 1) in the gel plaster group, placebo plaster was applied at the same acupoints in the placebo plaster group. The treatment was given from bedtime to early moming of the next day, 5 days were as one course, with 2-day interval, totally 4 courses were required in the both groups. Pittsburgh sleep quality index (PSQI), Epworth sleepiness scale (ESS) and Flinders fatigue scale were used to evaluate the sleep quality and fatigue level of the patients in the both groups before and after treatment and at 2 weeks of follow-up. The variations of insomnia TCM syndrome score and the 36-item short-form health survey (SF-36) score before and after treatment were observed.@*RESULTS@#Compared before treatment, the scores of PSQI, ESS and FFS after treatment and at follow-up were decreased in the both groups (@*CONCLUSION@#Acupoint application of gel plaster can effectively improve the quality of sleep and life in patients with insomnia.

Bibliometric analysis of refractory facial paralysis based on CNKI database / 中国针灸

The current development situation and the hotspot of the relevant research on refractory facial paralysis are explored. The articles on refractory facial paralysis are retrieved from CNKI database. The bibliographic items co-occurrence matrix builder (BICOMB) 2.0 is adopted to extract and analyze statistically literature characteristics and generate the high-frequency keywords matrix. The graphical clustering toolkit (gCLUTO) 1.0 is used to cluster the high-frequency keywords. A total of 750 articles are included, mostly published in

Antitumor effects of different Ganoderma lucidum spore powder in cell- and zebrafish-based bioassays / 中西医结合学报

OBJECTIVE@#Ganoderma lucidum spore (GLS) is gaining recognition as a medicinal part of G. lucidum and has been reported to possess various pharmacological properties, such as antitumor activity. In this work, wall-broken GLS powder (BGLSP) and wall-removed GLS powder (RGLSP), two kinds of GLS powder with different manufacturing techniques, were compared in terms of contents of active constituents and in vivo and in vitro antitumor effects.@*METHODS@#The ultraviolet and visible spectrophotometry method was used to determine the contents of polysaccharides and total triterpenoids in BGLSP and RGLSP. Seventeen individual triterpenoids were further quantified using ultra-high-performance liquid chromatography and quantitative analysis of multi-components by single marker. The antitumor effects of BGLSP and RGLSP were evaluated using in vitro cell viability assay against human gastric carcinoma SGC-7901, lung carcinoma A549 and lymphoma Ramos and further validated by in vivo zebrafish xenograft models with transplanted SGC-7901, A549 and Ramos.@*RESULTS@#The results showed that the contents of polysaccharides, total triterpenoids and individual triterpenoids of RGLSP were significantly higher than those of BGLSP. Although both BGLSP and RGLSP inhibited the three tumor cell lines in vitro in a dose-dependent manner, the inhibitory effects of RGLSP were much better than those of BGLSP. In the in vivo zebrafish assay, RGLSP exhibited more potent inhibitory activities against tumors transplanted into the zebrafish compared with BGLSP, and the inhibition rates of RGLSP reached approximately 78%, 31% and 83% on SGC-7901, A549 and Ramos, respectively.@*CONCLUSION@#The results indicated that the antitumor effects of GLS were positively correlated with the contents of the polysaccharides and triterpenoids and demonstrated that the wall-removing manufacturing technique could significantly improve the levels of active constituents, and thereby enhance the antitumor activity.

Microdialysis sampling and HPLC-MS/MS quantification of sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rat blood and brain extracellular fluid / 药学学报

The treatment plan for chronic pain often proceeds from a single drug to drug combination therapy. Sinomenine and ligustrazine, natural alkaline substances derived from traditional Chinese medicines, are expected to provide a new choice for combination analgesic therapy strategies. Here we establish a microdialysis sampling and HPLC-MS/MS quantification method for sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rat blood and brain extracellular fluid. Blood and brain microdialysis probes were implanted in the jugular vein toward the right atrium and left corpus striatum zone (AP +0.2 mm, ML 3.0 mm, DV 3.5 mm) in rats. The blood and brain microdialysis probes were perfused with citric acid buffer solution and Ringer's solution, respectively. Blood and brain extracellular fluid microdialysate were collected at intervals of 20 min at a perfusion rate of 1.5 μL·min-1, and continuously collected for 24 h after administration. The liquid chromatographic separation used a C18-reversed phase chromatographic column (HSS T3 2.5 μm, 2.1 mm×50 mm), the mobile phase was methanol/water (containing 0.05‰ formic acid), and gradient elution was carried out at a flow rate of 0.3 mL·min-1. Mass spectrometric detection used an electrospray ion source, positive ion mode and multi-reaction monitoring method. The selected quantitative ions for sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin, amitriptyline and internal standard naloxone were 330/181, 137/80, 172/154, 152/110, 160/142, 278/233 and 328/310 respectively. The specificity, linear range, matrix effect, accuracy, precision, stability and probe recovery were investigated and confirmed to be suitable for the determination of the above drugs in rat blood and brain extracellular fluid microdialysate. The calculated in vivo recovery of microdialysis probes ranged from 19.38% to 25.88%. After intravenous administration of sinomenine (50 mg·kg-1), ligustrazine (50 mg·kg-1), gabapentin (50 mg·kg-1), paracetamol (50 mg·kg-1), pregabalin (50 mg·kg-1) and amitriptyline (40 mg·kg-1) to rats, the peak concentration in the blood microdialysate was in the range of 0.2-10 μg·mL-1. Drug concentrations could also be detected in brain extracellular fluid microdialysate, however with lower levels (peak concentration: 0.1-6 μg·mL-1) than those of blood microdialysates at each time point. In conclusion, this method can be applied to microdialysis sampling and quantification of sinomenine, ligustrazine, gabapentin, paracetamol, pregabalin and amitriptyline in rats. The method will promote research in identifying herb-drug pharmacokinetic interactions, as well as safety concerns in combination-therapy strategies.

Protective Effect of Tiaogeng Decoction in Improving Apoptosis of Hypothalamic Neurons Based on JNK Pathway / 中国实验方剂学杂志

Objective:To study the mechanism of Tiaogeng decoction in improving apoptosis of hypothalamic neurons through c-Jun N-terminal kinase (JNK) pathway. Method:The GT1-7 cell line of hypothalamic neuron cells treated by tributyltin chloride (TBTC) was used to induce a model of neuronal apoptosis, with 17β-estradiol as a positive drug. They were divided into control group, model group (1 mg·L-1), 17β-E2 group(100 nmol·L-1) and low, middle and high-dose Tiaogeng decoction groups (31.25,62.5,125 mg·L-1). The cell morphology was observed under a fluorescent inverted microscope, and cell counting kit-8 (CCK-8) was used to detect the cell viability. The cell nuclear morphology and the apoptosis rate of hypothalamic neurons were detected by Hoechst 33258 and Annexin V-FITC/propidium iodide(PI) double staining. Western blot was used to detect the expression levels of apoptosis signal-regulating kinase 1(ASK1), JNK, p53 and cleaved Caspase-3 of JNK pathway in GT1-7 cell. Western blot was used to analyze apoptosis-associated protein apoptosis signal-regulated kinase 1 (ASK1), JNK, p53, cleaved Caspase-3 expressions in JNK pathway and phosphorylation of JNK after intervention with JNK inhibitor SP600125, protein expression level of cleaved Caspase-3. Result:Compared with normal control group, the cell viability of the model group was significantly decreased (PPPPP-1)treatment significantly inhibited the expressions of p-JNK and cleaved Caspase-3 in GT1-7 cells (PConclusion:Tiaogeng decoction can improve the apoptosis of hypothalamic neurons through JNK pathway, and provide a theoretical basis for the treatment of menopausal syndrome and central nervous system protection.

To improve gait of freezing in patients with Parkinson′s disease by electrical stimulation to common peroneal nerve delivered by wearable stimulator / 中华神经科杂志

Objective@#To explore whether the proprioceptive sensory cueing delivered by electrical stimulator to common peroneal nerve can improve the freezing of gait of parkinsonian patients.@*Methods@#Thirty patients with Parkinson′s disease experiencing freezing of gait (FOG) admitted to the First Affiliated Hospital of Anhui Medical University from January to December 2018 were included in the trial. Proprioceptive sensory cueing was provided by alternating electrical stimuli to bilateral common peroneal nerves delivered through the wearable electrical stimulator automatically triggered by walking. The modified 12 meters Timed Walking Test, six items of the modified Parkinson Activity Scale (PSA-6), and FOG score were used to test the gait function respectively when the stimulator was turned on and off.@*Results@#Compared to the off status, time duration for two 360° turns (T360), initiating (T1) and the turning (T2) was reduced with statistical significance when the stimulator was turned on in the three trial situations which were walking with no extra task (17.49 (13.55, 23.48) s vs 14.73 (10.31, 21.71) s, 2.16 (1.78, 2.68) s vs 1.70 (1.38, 2.29) s, 6.37 (4.10, 7.45) s vs 4.77 (3.40, 6.85) s; Z=-3.219, -4.206, -2.910, P<0.05), walking with cognitive task (21.35 (16.30, 30.72) s vs 18.36 (13.83, 27.98) s, 2.80 (2.05, 3.75) s vs 2.04 (1.64, 3.00) s, 6.58 (5.23, 8.96) s vs 5.75 (4.59, 7.76) s; Z=-3.486, -4.206, -3.363, P<0.05) and walking with motor task (25.34 (17.79, 30.30) s vs 22.24 (14.11, 29.33) s, 2.46 (2.19, 3.18) s vs 2.35 (1.66, 2.59) s, 7.77 (4.75, 9.93) s vs 6.45 (3.81, 7.66) s; Z=-3.468, -3.983, -3.570, P<0.05). In all the three exercise modes, the maintaining time (T3) was not significantly different. With the stimulator turned on, the total walking time (Tt) was not significantly different when the patients walked without extra task and with cognitive task but obviously improved with motor task (29.26 (20.11, 33.21) s vs 27.66 (17.70, 32.73) s, Z=-2.644, P=0.008). Compared to the off status, patients showed higher PAS-6 scores (18.99±2.55 vs 16.82±2.92, t=-6.617, P=0.000) and lower FOG scores (14.10±5.02 vs 10.61±5.05, t=6.151, P=0.000) with statistical significance when the stimulator was turned on.@*Conclusion@#The wearable electrical stimulator can alleviate FOG in patients with Parkinson′s disease by improving rotation, gait initiation and turning and may be used as a new rehabilitative therapy for patients with FOG.

To improve gait of freezing in patients with Parkinson′s disease by electrical stimulation to common peroneal nerve delivered by wearable stimulator / 中华神经科杂志

Objective To explore whether the proprioceptive sensory cueing delivered by electrical stimulator to common peroneal nerve can improve the freezing of gait of parkinsonian patients. Methods Thirty patients with Parkinson′s disease experiencing freezing of gait (FOG) admitted to the First Affiliated Hospital of Anhui Medical University from January to December 2018 were included in the trial. Proprioceptive sensory cueing was provided by alternating electrical stimuli to bilateral common peroneal nerves delivered through the wearable electrical stimulator automatically triggered by walking. The modified 12 meters Timed Walking Test, six items of the modified Parkinson Activity Scale (PSA?6), and FOG score were used to test the gait function respectively when the stimulator was turned on and off. Results Compared to the off status, time duration for two 360°turns (T360), initiating (T1) and the turning (T2) was reduced with statistical significance when the stimulator was turned on in the three trial situations which were walking with no extra task (17.49 (13.55, 23.48) s vs 14.73 (10.31, 21.71) s, 2.16 (1.78, 2.68) s vs 1.70 (1.38, 2.29) s, 6.37 (4.10, 7.45) s vs 4.77 (3.40, 6.85) s; Z=-3.219,-4.206,-2.910, P<0.05), walking with cognitive task (21.35 (16.30, 30.72) s vs 18.36 (13.83, 27.98) s, 2.80 (2.05, 3.75) s vs 2.04 (1.64, 3.00) s, 6.58 (5.23, 8.96) s vs 5.75 (4.59, 7.76) s; Z=-3.486,-4.206,-3.363, P<0.05) and walking with motor task (25.34 (17.79, 30.30) s vs 22.24 (14.11, 29.33) s, 2.46 (2.19, 3.18) s vs 2.35 (1.66, 2.59) s, 7.77 (4.75, 9.93) s vs 6.45 (3.81, 7.66) s; Z=-3.468,-3.983,-3.570, P<0.05). In all the three exercise modes, the maintaining time (T3) was not significantly different. With the stimulator turned on, the total walking time (Tt) was not significantly different when the patients walked without extra task and with cognitive task but obviously improved with motor task (29.26 (20.11, 33.21) s vs 27.66 (17.70, 32.73) s, Z=-2.644, P=0.008). Compared to the off status, patients showed higher PAS?6 scores (18.99±2.55 vs 16.82±2.92, t=-6.617, P=0.000) and lower FOG scores (14.10 ± 5.02 vs 10.61 ± 5.05, t=6.151, P=0.000) with statistical significance when the stimulator was turned on. Conclusion The wearable electrical stimulator can alleviate FOG in patients with Parkinson′s disease by improving rotation, gait initiation and turning and may be used as a new rehabilitative therapy for patients with FOG.

UHPLC quantification and brain permeability study of sinomenine and ligustrazine in CQM after single intravenous administration / 药学学报

Chuanxiong Qingfengteng mixture (CQM) is an analgesic developed based on clinical evidence and traditional Chinese medicine theory, which majorly consists of Ligusticum chuanxiong and Sinomenium acutum extracts. The current study aims to establish an UHPLC-UV method for the quantification of sinomenine and ligustrazine after CQM administration to rats, mice and cells, and to study the brain permeability of sinomenine and ligustrazine. The selectivity, linearity, accuracy, precision and stability of the established method demonstrated that it was suitable for the determination of sinomenine and ligustrazine in biological samples such as plasma, brain tissue and cellular fluid. After CQM was intravenously administered to rats and mice, both sinomenine and ligustrazine were detected in the brain from 5 min-2 h. The CSF/plasma partition coefficients (Kp, C/P) of each component were higher than those of brain tissue/plasma partition coefficient (Kp, B/P), the Kp, C/P and Kp, B/P of ligustrazine were higher than those of sinomenine. The concentrations between CSF and brain tissue were strongly correlated (Pearson's R>0.86, P<0.001). The unbound fraction in plasma of sinomenine and ligustrazine was 78.92% and 34.07%, respectively. The plasma protein binding rates displayed concentration-independent behavior within their respective in vivo concentration ranges. After CQM co-cultured with Caco-2 cell monolayers, the apparent permeability coefficient (Papp) of sinomenine and ligustrazine were 1.30×10-6 and 3.64×10-6 cm·s-1, respectively, following into the range of the intermediate and high permeability compounds. The efflux ratio (Papp(basolateral→apical)/Papp(apical→basolateral)) of sinomenine and ligustrazine were 0.67 and 0.85, respectively. When combined with P-glycoprotein inhibitor, the Papp of each component did not increase. In conclusion, the UHPLC-UV assay was successfully applied for the brain permeability study of CQM, the components of CQM can be quickly distributed to cerebrospinal fluid and pass through the blood-brain barrier. The brain permeability of ligustrazine is higher than that of sinomenine. The transmembrane transport of sinomenine and ligustrazine may not be affected by efflux transporters. All animal care and use complied with the Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Council of the People's Republic of China. All animal studies were implemented according to protocols, which were reviewed and approved by the Institutional Animal Care and Use Committee at Experimental Research Center, China Academy of Chinese Medical Sciences.

Study on expression, role and mechanism of leukocyte adhesion molecule CD44 in pulmonary tuberculosis patients / 中国免疫学杂志

Objective: To study the expression of CD44 in pulmonary tuberculosis patients and explore the possible mechanisms. Methods: 236 patients suspected with pulmonary tuberculosis were selected and divided into pulmonary tuberculosis group ( n= 152) and non-pulmonary tuberculosis group ( n= 84), and 100 healthy people were randomly selected as healthy control group. The expression of CD44 was evaluated by qRT-PCR and ELISA in peripheral blood and pleural effusion from different patients. The CD44 levels at pre and post-treatment time points were determined by ELISA. The inducing factors of increased CD44 and the potentialroles played by CD44 in the pathogenesis of TB were also evaluated. We elucidated whether CD44 detection could combine with T-spot. TB to diagnose pulmonary tuberculosis rapidly and accurately. Results: The expression of CD44 in pulmonary tuberculosis patients was higher compared with the non-pulmonary tuberculosis patients and healthy people, and would be down-regulated after treatment for 3, 6 and 9 months. Besides, CD44 could not remove H37 Ra by the CFU assay, and could promote the expression of CCL-2, indicating that CCD4 promote the mobility the THP-1 via induction of CCL-2. Besides, TNF-α neutralizing antibody, added into the macrophages, could inhibit the expression of CD44, and functional TNF-α induced the expression of TNF-α. Conclusion: CD44 is highly expressed in pulmonary tuberculosis patients, which may be due to the high expression of TNF-α in pulmonary tuberculosis patients, stimulating macrophages to produce CD44, and it will provide a basis for clinical diagnosis of pulmonary tuberculosis.

Stroke, platelet activating factor and receptor antagonists / 中国中药杂志

Stroke is an acute cerebrovascular disease with high morbidity, disability and mortality. The prevention and treatment conditions for stroke is severe all over the world. Antiplatelet aggregation is an effective treatment. Platelet activation factor (PAF) is another important medium in mediating platelet aggregation, which plays an important role in the pathogenesis of stroke. In recent years, PAF receptor antagonists have attracted international attention in the field of stroke prevention and treatment. In this review, we would summarize the classification, mechanism and drug characteristics of PAF receptor antagonists in order to provide the valuable guidance and direction for clinical medicine and research.

Analgesic effect and central mechanisms of CQ prescription on cancer invasion induced mirror image pain in model mice / 中国中药杂志

This study aimed to analyze the analgesic effect and related central mechanisms of CQ prescription on cancer invasion induced mirror image pain (CIIMIP)in model mice.In the study, male BALB/c mice were randomly divided into normal group, operation control group (injected with 0.2 mL inactivated S180 sarcoma cell sap), model group (injected with 0.2 mL S180 sarcoma cell sap on the right leg near the greater trochanter of femur) and CQ prescription low dose group (intraperitoneally injected with CQ prescription 100 mg•kg⁻¹ on the basis of model mice), CQ prescription middle dose group (intraperitoneally injected with CQ prescription 150 mg•kg⁻¹ on the basis of model mice), and CQ prescription high dose group (intraperitoneally injected with CQ prescription 200 mg•kg⁻¹ on the basis of model mice). Mechanical withdraw threshold (MWT) of the mirror image lateral hind paws were evaluated by Von Frey hairs before modeling and after surgery. The levels of glutamate (Glu), gamma aminobutyric acid (GABA), glycine (Gly), and taurine (Tau) in the L3-L5 spinal cord were measured by the high performance liquid chromatography-fluorescence detector (HPLC-FLD); AimPlex detection technology with multiple factors was used to detect the levels of regulated on activation in normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein (MCP-3) in the L3-L5 spinal cord. Then we observed the influence of GABAa receptor antagonist (Bicuculline) on analgesic effect of CQ prescription.The results indicated that CQ prescription could remarkably increase MWT of model mice(P<0.01, P<0.05), decrease the level of Glu(P<0.01, P<0.05), improve the levels of GABA, Gly, Tau(P<0.01, P<0.05), lower the ratio of Glu/GABA(P<0.01, P<0.05), and reduce the levels of RANTES, MCP-3(P<0.05) in the L3-L5 spinal cord, and GABAa receptor antagonist significantly blocked the analgesic effect of CQ prescription at two time points(P<0.05).This study showed that CQ prescription had significant analgesic effect on CIIMIP model mice, and its mechanism was associated with regulating the balance between excitability amino acid(EAA) and inhibitory amino acid (IAA) transmitters in central nervous system, partially activating GABAa receptor, and reducing the release of RANTES and MCP-3 in the spinal cord.

Transcutaneous vagus nerve stimulation for primary insomnia and affective disorder:a report of 35 cases / 中国针灸

<p><b>OBJECTIVE</b>To observe and evaluate the clinical efficacy of transcutaneous vagus nerve stimulation (taVNS) at auricular concha for primary insomnia (PI) and affective disorder.</p><p><b>METHODS</b>A total of 35 patients who met the diagnosis standard of PI in(5th edition) were included. The self-developed auricular vagus nerve stimulator (TENS-200A) was applied at auricular concha, 30 min per treatment, twice a day, 5 days a week for consecutive 4 weeks. The follow-up visit was conducted at the end of 6th week. The Pittsburg sleep quality index scale (PSQI), 17-items Hamilton depression scale (17HAMD) and Hamilton anxiety scale (HAMA) were applied for evaluation. The PSQI, HAMA and 17HAMD were observed before and after treatment; the safety was also observed.</p><p><b>RESULTS</b>Compared before treatment, the PSQI was significantly decreased to (13.20±3.61) at the end of 2nd week (<0.05); compared before treatment, the 17HAMD and HAMA were significantly decreased at the end of 4th week and 6th week (all<0.05). No adverse reaction was observed.</p><p><b>CONCLUSIONS</b>The taVNS could not only relieve PI symptoms, but also improve the depressive and anxiety symptoms, in addition, it may have positive long-term efficacy and safety.</p>

Study on relationship between constitution and syndrome of MMP-9, IL-6 and MTHFR gene in patients with ischemic stroke / 中国中药杂志

By studying the relationship between syndromes, physique and MMP-9, IL-6 and MTHFR gene polymorphisms in patients with ischemic stroke,The relationship between MMP-9, IL-6 and MTHFR gene polymorphism was analyzed in patients with ischemic stroke.The data were collected by collecting the data of patients with ischemic stroke, and the statistical analysis was carried out. Syndrome：61 cases of ischemic stroke patients with stroke phlegm stasis syndrome in patients with the highest frequency, a total of 30 cases; Physical constitution： phlegm is ischemic stroke patients prone to physical, a total of 20 cases; The analysis of the relationship between constitution and syndrome shows that the patients with qi deficiency constitution tend to show qi deficiency and blood stasis syndrome after onset, The analysis of the relationship between constitution and syndrome shows that the patients with qi deficiency constitution tend to show qi deficiency and blood stasis syndrome after onset, Phlegm constitution and physical condition after the onset of symptoms tend to wind phlegm stasis syndrome; Syndrome and MMP-9, IL-6 relationship：The distribution of MMP-9 and IL-6 in patients with qi and phlegm stasis syndrome and qi deficiency and blood stasis syndrome was significantly different from that in Z test (P<0.05). The level of MMP-9 in patients with qi deficiency and blood stasis syndrome was significantly higher than that in patients with wind phlegm and blood stasis syndrome;The level of IL-6 in patients with phlegm and blood stasis syndrome was significantly higher than that in patients with qi deficiency and blood stasis syndrome. Syndrome, constitution and MTHFR gene polymorphism： among the 61 samples, 34 were heterozygous mutations, 15 were pure and mutated, 12 had no mutation, The mutation rate of this locus was 4.08 times that of patients without mutations.The genotype of MTHFR C677T in patients with phlegm dampness tends to be CT genotype. Wind phlegm stasis syndrome in patients with easy to appear after the TT genotype; Yin deficiency syndrome in patients prone to miscellaneous and mutations, the performance of CT genotype; Analysis of the relationship between syndromes and physique in patients with ischemic stroke,Phlegm and dampness, flat quality patients after the onset of easy to show the wind phlegm stasis syndrome; Qi deficiency after the onset of symptoms in patients with Qi and blood stasis. Suggesting that before the onset of such as for the partial physical conditioning, may be on the prevention of ischemic stroke have a certain effect; Analysis of the relationship between syndromes and MMP-9 and IL-6 in patients with ischemic stroke, Wind phlegm stasis syndrome and IL-6 levels are related, Qi deficiency and blood stasis syndrome and MMP-9 levels are related. Analysis of the relationship between syndromes and MTHFR gene polymorphism in patients with ischemic stroke, TT genotype after the onset of symptoms prone to wind phlegm stasis syndrome, CT genotype patients after the onset of easy manifestations of Yin deficiency wind syndrome; Analysis of the relationship between physique and MTHFR gene polymorphism in patients with ischemic stroke, CT genotype is easy to show phlegm.For more in-depth understanding of pathogenesis of ischemic stroke to provide the basis, For the clinical treatment and prevention to provide intervention strategies.

Inhibitory Effect of Feiji Recipe on IDO Induced Immune Escape on the Murine Model of Lewis Lung Carcinoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Feiji Recipe (FR) intervening indoleamine 2,3-dioxygenase (IDO) induced immune escape on the murine model of Lewis lung carcinoma. Methods Totally 48 C57BL/6 mice inoculated with Lewis lung cancer cells transfected with human (enhanced green fluorescent protein,EGFP)-IDO gene were divided into four groups according to radom digit table, i.e., the model group (administered with normal saline by gastrogavage) , the Chinese medicine group (treated with FR Decoction at the daily dose of 100 mg/g by gastrogavage), the 1-methyl-D-trytaphan (1-MT) group (administered with 1-MT mixed liquor at the daily dose of 100 mg/kg by gastrogavage), and the Paclitaxel group (treated with Paclitaxel at the daily dose of 15 mg/kg by peritoneal injection), 12 in each group. The intervention was started from the 2nd day of modeling. The survival time was observed in 24 of them. Ratios of CD4+ CD25+ FoxP3+ regulatory T cells (Treg) in the spleen were detected in the rest 24 mice by flow cytometry respectively.</p><p><b>RESULTS</b>Compared with the model group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells remarkably decreased in the Chinese medicine group, the 1-MT group, and the Paclitaxel group (P < 0. 01). Compared with the Paclitaxel group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells decreased significantly in the 1-MT group (P < 0.05).</p><p><b>CONCLUSION</b>FR could inhibit the proliferation of lung cancer cells and immune eseape, improve the immune function, and prolong the survival of tumor-bearing mice.</p>

Cut-off Values of Diagnostic Indices to Detect Iron Deficiency in Chinese Breast-fed Infants / 生物医学与环境科学（英文）

Iron deficiency anemia is one of the most prevalent nutritional deficiency worldwide. The commonly used cut-off values for identifying iron deficiency are extrapolated from older children and may not be suitable for infants. Therefore, our study aimed to establish appropriate cut-off values for the evaluation of iron status in Chinese infants. Pregnant women who delivered at ⋝37 gestational weeks with normal iron status were recruited. Later, infants with normal birth weight and who were breastfed in the first 4 months were selected. Blood samples were collected to assess hemoglobin, serum ferritin, soluble transferrin receptor, mean corpuscular volume and free erythrocyte protoporphyrin. Cut-offs of all iron indices were determined as the limit of 95% confidence interval.