Study on the objectivity and biological mechanism of Psoralea corylifolia Linn.'s 'Great dryness damages the liver' / 药学学报

According to the theory of 'Xingben Dazao' of Psoralea corylifolia Linn. (BL), the susceptible syndromes and biomarkers of liver injury caused by BL were searched. Rat models of kidney-yin deficiency syndrome (M_yin) and kidney-yang deficiency syndrome (M_yang) were established, and all animal experimental operations and welfare following the provisions of the First Affiliated Experimental Animal Ethics and Animal Welfare Committee of Henan University of Traditional Chinese Medicine (No. YFYDW2020017). The results showed that BL significantly decreased the body weight, water intake, and urine weight of M_yin rats and increase the organ indexes of the liver, testis, adrenal gland, and spleen and the expression of alanine aminotransferase (ALT). Meantime, BL significantly increased the urine weight of M_yang rats and decreased the expression of ALT and aspartate aminotransferase (AST). Hematoxylin and eosin (HE) staining showed that BL could aggravate inflammatory infiltration of hepatocytes in rats with M_yin and alleviate liver injury in rats with M_yang. Metabolomics identified 17 BL co-regulated significant differential metabolic markers in M_yin and M_yang rats. Among them, 8 metabolites such as glutamine, quinolinate, biliverdin, and lactosylceramide showed opposite trends, mainly involving cysteine and methionine metabolism, tyrosine metabolism, tryptophan metabolism, purine metabolism, sphingolipid metabolism, glycerol phospholipid metabolism, glutamine metabolism, and other pathways. M_yin/M_yang may be the susceptible constitution of BL for liver damage or protection, which may be related to the regulation of amino acid metabolism and sphingolipid metabolism. The study can provide some experimental data support for the safe and accurate use of BL in the clinical practice of traditional Chinese medicine.

Identification of components in vitro and plasma of Jinhua Qinggan granules and preliminary pharmacokinetics analysis / 药学学报

As one of the "Three Drugs Three Prescriptions" anti-COVID-19 traditional Chinese medicine, Jinhua Qinggan granules (JHQG) has been proved to have clear clinical effects. With complex medicinal flavors and ingredients, there is no systematic research report on chemical composition in vivo or in vitro. An ultrahigh pressure liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF/MS) method was developed in this study to identify the components of the anti-COVID-19 traditional Chinese medicine JHQG granules. Analyze the collected rat plasma samples after administration and explore the exposed components in rats within 8 hours after intragastric administration. Preliminary pharmacokinetic analysis was then performed on this basis. Through UPLC-QTOF/MS analysis and verification by standard products, a total of 77 chemical components in JHQG formula have been identified, among which 22 compounds were highly exposed in vivo, mainly derived from three medicinal materials of honeysuckle, scutellaria and forsythia. Through the assessment of the blood drug concentration by the compartment model, 6 PK parameters of 4 high-exposure chemical components have been obtained, clarifying the metabolic characteristics of the main exposed components in JHQG briefly. The method is simple, efficient, sensitive and accurate and provides research basis to the clarification of the pharmacodynamics material basis and mechanism of JHQG, which has certain reference significance for the basics and applications research of the traditional Chinese medicine prescriptions in fighting the SARS-CoV-2.

Study on alleviating effect of Glycyrrhizae Radix et Rhizoma on Psoraleae Fructus-induced liver injury based on network pharmacology and cell experiments / 中国中药杂志

This study was designed to explore the alleviating effect and mechanism of Glycyrrhizae Radix et Rhizoma against Psora-leae Fructus-induced liver injury based on network pharmacology and cell experiments. The active components of Glycyrrhizae Radix et Rhizoma and Psoraleae Fructus were first retrieved from the Encyclopedia of Traditional Chinese Medicine(ETCM), Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), Comparative Toxicogenomics Database(CTD), and literature and further screened by SwissADME. The obtained 25 potential toxic components of Psoraleae Fructus and 29 flavonoids in Glycyrrhizae Radix et Rhizoma were input into the SwissTargetPrediction for target predication. A total of 818 targets related to liver injury were screened out based on GeneCards and MalaCards, and 91 common targets of Psoraleae Fructus, Glycyrrhizae Radix et Rhizoma, and liver injury were obtained from Venny. STRING was applied for constructing the PPI network, and Metascape for analyzing the biological processes and signaling pathways that common targets participated in. Cytoscape was used to construct the component-target-disease network and component-target-pathway network for Glycyrrhizae Radix et Rhizoma against Psoraleae Fructus-induced liver injury. The predicted core targets were proto-oncogene tyrosine-protein kinase(SRC), phosphatidylinositol 4,5-bisphosphate 3-kinase subunit alpha(PIK3 CA), RAC-alpha serine/threonine-protein kinase(AKT1), etc, with PI3 K-AKT signaling pathway, MAPK signaling pathway, apoptosis, Toll-like receptor signaling pathway, and NF-κB signaling pathway mainly involved. Following the scree-ning of the main toxic and pharmacodynamic components, the pharmacodynamic effects were investigated by cell experiments. The results showed that licochalcone A was mainly responsible for alleviating coryfolin-induced liver injury, licochalcone B for coryfolin-and psoralidin-induced liver injury, and echinatin for corylifolinin-and bakuchiol-induced liver injury. The preliminary revealing of the alleviating effect of Glycyrrhizae Radix et Rhizoma on Psoraleae Fructus-induced liver injury and the prediction of related mechanisms will provide reference for further mechanism research and reasonable clinical compatibility.

Translocator protein mediated anti-PTSD effects of YL-IPA08 / 军事医学

Objective To explore the role of 18 ku translocator protein (TSPO) in the anti-post-traumatic-stress-disorder(PTSD) effects of YL-IPA08 and the value of TSPO as a potential pharmacological target using gene knock out mice.Methods The PCR method was used to genotype TSPO wild type (WT) mice and knock out (KO) mice.Foot shock was used to establish a well-accepted mouse model of PTSD,the open field test (OFT) was used to evaluate the locomotor activity in mice,and freezing measurement was used to evaluate the PTSD-like fear behavior in mice.Results Compared with TSPO WT mice,KO mice had no expressible TSPO gene,but showed similar locomotor activity to WT mice after PTSD modeling.On day 1,day 5 and day 16 after PTSD modeling (day-1-day 0),both WT and KO mice showed significant PTSD-like behavior with enhanced freezing time.However,8 d treatment (day 0-day 7) of YL-IPA08 (0.3 mg/kg,once daily) or positive drug sertraline (15 mg/kg,once daily) after PTSD modeling significantly reduced freezing time selectively in WT mice,but not in KO mice.Conclusion It has been found for the first time that TSPO WT and KO mice can show the same sensitivity to PTSD modeling (namely the same PTSD-like behavior performance).Interestingly,TSPO can mediate the anti-PTSD effects of YL-IPA08.Therefore,the present study provides direct evidence for the value of TSPO as an potential pharmacological target for PTSD.

Correlation of in vivo and in vitro methods in measuring choroidal vascularization volumes using a subretinal injection induced choroidal neovascularization model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In vivo quantification of choroidal neovascularization (CNV) based on noninvasive optical coherence tomography (OCT) examination and in vitro choroidal flatmount immunohistochemistry stained of CNV currently were used to evaluate the process and severity of age-related macular degeneration (AMD) both in human and animal studies. This study aimed to investigate the correlation between these two methods in murine CNV models induced by subretinal injection.</p><p><b>METHODS</b>CNV was developed in 20 C57BL6/j mice by subretinal injection of adeno-associated viral delivery of a short hairpin RNA targeting sFLT-1 (AAV.shRNA.sFLT-1), as reported previously. After 4 weeks, CNV was imaged by OCT and fluorescence angiography. The scaling factors for each dimension, x, y, and z (μm/pixel) were recorded, and the corneal curvature standard was adjusted from human (7.7) to mice (1.4). The volume of each OCT image stack was calculated and then normalized by multiplying the number of voxels by the scaling factors for each dimension in Seg3D software (University of Utah Scientific Computing and Imaging Institute, available at http://www.sci.utah.edu/cibc-software/seg3d.html). Eighteen mice were prepared for choroidal flatmounts and stained by CD31. The CNV volumes were calculated using scanning laser confocal microscopy after immunohistochemistry staining. Two mice were stained by Hematoxylin and Eosin for observing the CNV morphology.</p><p><b>RESULTS</b>The CNV volume calculated using OCT was, on average, 2.6 times larger than the volume calculated using the laser confocal microscopy. The correlation statistical analysis showed OCT measuring of CNV correlated significantly with the in vitro method (R 2 =0.448, P = 0.001, n = 18). The correlation coefficient for CNV quantification using OCT and confocal microscopy was 0.693 (n = 18, P = 0.001).</p><p><b>CONCLUSIONS</b>There is a fair linear correlation on CNV volumes between in vivo and in vitro methods in CNV models induced by subretinal injection. The result might provide a useful evaluation of CNV both for the studies using CNV models induced by subretinal injection and human AMD studies.</p>

125I brachytherapy combined with chemotherapy for patients with inoperable stage Ⅲ non-small cell lung cancer / 中华核医学杂志

Objective To assess the therapeutic effect of radioactive seed 125I brachytherapy combined with GP chemotherapy regimen (gemcitabine 1000 mg/m2, cisplatin 75 mg/m2) for inoperabale stage Ⅲ non-small cell lung cancer (NSCLC). Methods Thirty-nine documented inoperable stage Ⅲ NSCLC patients, enrolled between January 2005 and June 2008 for the study group, were treated with the combination of 125I brachytherapy and GP regimen. The brachytherapy methods were conducted according to TPS and each patient was treated under those patients were treated with standard GP regimen. Chest CT scans were performed every three months post-procedurally, until disease progression or recurrence. The follow-up time was up to twenty four months after treatment. In the control group, equal amount of Ⅲ stage NSCLC patients were treated with standard GP regimen alone. Chi-square test and survival analysis with Kaplan-Meier and Log-rank were used to compare the differences of recent (3 months after therapy)efficiency, survival rate, survival time between two groups. Results The re-cent effective rates of the study group (71.8%, 28/39) and control group (61.5%, 24/39) were not statistical different (χ2=0.93, P>0.05), yet the tumor CR rates in two groups showed significant disparity (χ2=4.48, P0.05). However, singificant differences (χ2=4.07, 4.63,both P<0.05) were found in 2-year survival rate and median survival time, with 41.0% (16/39) vs 23.1% (9/39) and 18.9±2.7 months vs 14.2±0.7 months. Conclusions 125I brachytherapy combined with GP regimen chemotherapy could be an effective treatment method and could improve the tumor CR rate and survival rate for patients with inoperable stage Ⅲ NSCLC.

The relationship between the expression of NF-kB, TGFbeta1, FN and hepatic fibrosis in diabetic rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression of nuclear factor kappa B (NF-kB), transforming growth factor beta1 (TGFbeta1), fibronectin (FN) in liver from diabetic rats.</p><p><b>METHODS</b>Twenty male Sprague-Dawley rats were divided randomly into two groups: normal control group (n = 10) and type 2 diabetic group (n = 10). After 4 weeks of high-fat feeding, diabetic group rats were injected with low dosage streptozotocin (30 mg/kg) intraperitoneally to induce type 2 diabetic rat models. The diabetic rats received high-fat feeding for another 12 weeks. At the end of the experiment, the fibrosis lesion was observed under light microscopy after Masson staining. The mRNA levels of NF-kB, TGFbeta1, FN from rats liver were assayed by semi-quantity RT-PCR, the protein levels of NF-kB, TGFbeta1, FN was detected by IHC.</p><p><b>RESULTS</b>Fibrosis was found in diabetic rats. The levels of TGFbeta1, FN mRNA in liver tissues increased in diabetic rats compared with normal control rats (0.91+/-0.19 vs 0.47+/-0.20, t = 5.233, P less than 0.05; 1.85+/-0.70 vs 1.22+/-0.39, t = 2.463, P less than 0.05). And the protein levels of NF-kB P65, TGFbeta1, FN in liver tissues from diabetic rats were significantly higher than those in normal control rats (10978.77+/-8782.59 vs 4206.86+/-1430.56, Z = 1.979, P less than 0.05; 8551.00+/-4768.68 vs 4036.85+/-1051.12, Z = 2.303, P less than 0.05; 16980.30+/-11529.29 vs 5701.95+/-9461.75, t = -2.391, P less than 0.05).</p><p><b>CONCLUSION</b>Upregulation of NF-kB, TGFbeta1, FN in liver tissues may play a role in the hepatic fibrogenesis in diabetic rats.</p>

Distribution characteristics of ApoB3'VNTR and hyperlipemia correlation factors in civil aircrew / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the frequency distribution in polymorphism of the apoprotein B 3' variable number tandem repeat (ApoB3'VNTR) and influence factors on hyperlipemia in civil aircrew.</p><p><b>METHODS</b>ApoB genotypes were determined by PCR technology and agarose gel electrophoresis. The blood lipids were measured by routine kits. Personal information of flight personnel was collected by questionnaire.</p><p><b>RESULTS</b>Prevalence of the total dyslipidemia (49.5%) and overweight (55.6%) of flight personnel were much higher than that of domestic general population (29.2% and 49.1%) respectively (P < 0.05). There were 16 alleles and 54 kinds of genotypes of ApoB3'VNTR in the 682 flight personnel. The frequency distribution of alleles and genotypes of aircrew in the two air companies had same trend, which was different from the general population. The frequency of the homozygote was 76.54%, which was much higher than that of the other peoples home and abroad (21.50%). The frequency of the big allele (VNTR > or =39) in hyperlipemia groups were higher than that of normal groups. By analysis of co-variance, the body mass index (BMI), low density lipoprotein (LDL) and the total cholesterol(TC) increased with the cumulate flight hours (P < 0.05). In multivariate logistic regression analysis, the BMI was the only factor influencing blood lipids, and the cumulate flight hours was only factors affecting the BMI. Taking the cumulate flight hours logarithm as the independent variable(X), and the BMI as dependent variable(Y), the linearity equations was: Y = 2.730X + 13.584 (R2 = 0.159, P < 0.01).</p><p><b>CONCLUSION</b>There are perhaps special genetic characteristics in the polymorphism of the ApoB3'VNTR in the aircrew. The big allele is correlated with the hyperlipemia. The flight burden not only directly affects the BMI and blood lipids levels, but also it can indirectly affect the lipids levels by BMI.</p>

Serum-free Aggregate Perfusion Culture of CHO Cells:an Ultrasonic and Sedimentation Column Combined Perfusion System / 中国生物工程杂志

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.

Quantitation of albumin in urine by HPLC and initial clinical application / 中华检验医学杂志

Objective To measure albumin in urine by HPLC and conduct primary clinical application Methods Solvent gradient and appropriate wave length was optimized and performance of the HPLC method was evaluated.Urine albumin of 46 patients with diabetes was measured.Results In standard and urine,retention time of Alb was 13.1 min.The linear measuring range extends to 1 820 mg/L.The lower limit of measurment for Alb was 4.2 mg/L.The intra-assay CV and the inter-assay CV were 3.36%,4.12% and 1.93%,1.97% at 24.5 mg/L and 546.9 mg/L of Alb respectively.Analytical recovery rate were 96.3%,98.2% and 97.5%.Microalbuminuria rate was 54.3% by HPLC,26.1% by immunoassay in 46 patients with diabetes.Conclusions Measurement of Alb in urine by HPLC is feasible as routine method until quantifying urinary total Alb conveniently.HPLC is the same to suit research for diabetic nephropathy and so on.