Ag85B with c-di-AMP as mucosal adjuvant showed immunotherapeutic effects on persistent Mycobacterium tuberculosis infection in mice

Tuberculosis (TB), caused by Mycobacterium tuberculosis, remains the leading cause of mortality by a single infectious agent in the world. M. tuberculosis infection could also result in clinical chronic infection, known as latent TB infection (LTBI). Compared to the current limited treatment, several subunit vaccines showed immunotherapeutic effects and were included in clinical trials. In this study, a subunit vaccine of Ag85B with a novel mucosal adjuvant c-di-AMP (Ag85B:c-di-AMP) was delivered intranasally to a persistent M. tuberculosis H37Ra infection mouse model, which also presented the asymptomatic characteristics of LTBI. Compared with Ag85B immunization, Ag85B:c-di-AMP vaccination induced stronger humoral immune responses, significantly higher CD4+ T cells recruitment, enhanced Th1/Th2/Th17 profile response in the lung, decreased pathological lesions of the lung, and reduced M. tuberculosis load in mice. Taken together, Ag85B:c-di-AMP mucosal route immunization provided an immunotherapeutic effect on persistent M. tuberculosis H37Ra infection, and c-di-AMP, as a promising potential mucosal adjuvant, could be further used in therapeutic or prophylactic vaccine strategies for persistent M. tuberculosis infection as well as LTBI.

Hemolytic diseases of the fetus and newborn caused by anti-Dib: a case report and related research / 中国输血杂志

【Objective】 To identify the specificity of alloantibody against high-frequency antigens in one case suffering with severe hemolytic diseases of the fetus and newborn (HDFN) and to screen for matching blood for transfusion. 【Methods】 The HDFN test and the antibody serological identification tests in the mother were performed. Several common high frequency antigens of maternal red blood cells (RBCs) were determined. IgG subtype coated on the RBCs of the newborn was determined. The phagocytic efficiency of the antibody was tested using the monocyte phagocytosis of sensitized erythrocyte by flow cytometry in vitro. Sanger sequencing of DI gene was performed in the mother, father and mother’s brother. The diluted maternal plasma was used for large scale screening of matching blood using IAT in Coomb’s gel card. 【Results】 Di(b-) phenotype was identified in the mother of the newborn and anti-Dib (titer: 512) related HDN was detected in the newborn. IgG1 and IgG2 subtypes of anti-Dib were detected and the rate of monocyte phagocytosis was 88.83%(74.7/84.09). The compatible blood was not detected in the maternal relatives. Subsequently, the newborn received the matching RBCs of two Di(b-) donors identified from 5 520 blood donors and discharged from the hospital. We screened out 17 Di(b-) donors out of 51 334 blood donors, indicating that the distribution frequency of Di(b-) among blood donors in Guangzhou was about 0.033% (17/51 334). 【Conclusion】 By serology and molecular biology methods, the newborn was identified with HDFN caused by anti-Dib, and an effective large-scale screening method for Di (b -) rare blood types was established to find matching blood, which supported the establishment of rare Di(b-) blood database.

Clinical characteristics and related factors of patients with schizophrenia in different ages of onset / 中国心理卫生杂志

Objective:To explore the clinical characteristics and related socio-demographic factors of schizo-phrenia patients with different ages of onset.Methods:Totally 2 016 patients with schizophrenia aged 15 to 70 were selected according to the diagnostic criteria for schizophrenia in the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition.All of the patients were interviewed by psychiatrists using the Mini International Neuropsy-chiatric Interview to diagnose schizophrenia,Clinical-Rated Dimensions of Psychosis Symptom Severity(CRDPSS)and the Positive and Negative Syndrome Scale(PANSS)to assess symptoms.The cut-off points were 18 and 25 years old for three age groups,i.e.early onset(EOS),youth onset(YOS)and adult onset(AOS).Statistical analy-ses were performed by analysis of variance Pearson correlation analysis,and multivariate linear regression.Results:The early-onset patients had the highest total PANSS score(73.8±28.0)and CRDPSS score(11.7±5.4).Fe-male gender,high education level,Han ethnicity,early onset age,and slower onset of illness were negatively corre-lated with the total and dimension score of PANSS scale and CRDPSS scale(standardized regression coefficient:0.04-0.47),and income level and smoking were negatively correlated with those score(standardized regression coefficient:-0.04--0.14).Conclusion:Early-onset schizophrenia patients have more severe symptoms,and fe-male,high education level,early-onset disease,and chronic onset are the risk factors of symptom severity in patients with schizophrenia.

Discussion on the Treatment of Skin Diseases with Warming Yang Method Based on the Theory of"Balance between Yin and Yang"in Huang Di Nei Jing / 中国中医药信息杂志

"Balance between yin and yang,the spirit is treated"means the best health state of the human body.The theory of yin and yang is the foundation of TCM theory,and the key to the treatment of chronic skin diseases is to regulate yin and yang.The key to the balance of yin and yang lies in the sufficiency of yang-qi.Sufficient yang-qi can guarantee the defense outside is solid and dense.The healthy-qi inside can avoid the invasion of pathogenesis.The invisible pathogenesis is difficult to transform into tangible pathogenesis.In the clinical syndromes,it has been found that the lack of yang-qi is one of the important reasons for the occurrence and development of skin diseases.By warming yang and dispersing pathogenic factors,dissolving muscles and dispelling wind,reconciling nutrient and defense,warming and dissipating cold,clearing heat and detoxifying,nourishing blood and moistening dryness,etc.,the deficiency yang can be warmed,the yang can be concealed,the balance between yin and yang can be achieved,and the diseases can be eliminated.

Toxicity and Mechanism of Di-(2-ethylhexyl) Phthalate on Testis / 生物化学与生物物理进展

Di-(2-ethylhexyl) phthalate (DEHP) is currently one of the most widely used plasticizers, widely found in all kinds of items, such as children’s toys and food packaging materials, but also added to wallpaper, cable protective agents and other building decoration materials. DEHP is toxic and absorbed by the human body through respiratory tract, digestive tract and skin contact, which can cause damage to multiple systems, especially the male reproductive system, and testis is an important target organ. Oxidative stress injury is the core mechanism of spermatogenesis disorder caused by DEHP. DEHP exposure can cause oxidative stress or reactive oxygen species (ROS) increase in germ cells, and on this basis, promote cell apoptosis or cause excessive autophagy. The toxicity of DEHP to Leydig cells is mainly to interfere with the synthesis of steroid hormones. For Sertoli cells, ferroptosis and destruction of the blood-testis barrier are common injury mechanisms. In addition, gene methylation caused by DEHP not only affects the spermatogenic process, but also has epigenetic effects on offspring. In this paper, we reviewed the pathological damage, germ cell toxicity and epigenetic effects of DEHP on testis, and focused on the damage and molecular mechanism on testicular spermatogenic cells, Leydig cells and Sertoli cells. Future research is required to elucidate the body’s clearance mechanism and treatment plan after exposure to DEHP and whether DEHP will damage the function of myoid cells. It is hoped that this can provide new ideas for prevention and treatment of male reproductive disorders resulting from long-term exposure to plastic products.

Mechanism of bis（2-ethylhexyl） phthalate in inducing cholestasis and liver injury in mice / 临床肝胆病杂志

ObjectiveTo investigate the mechanism of bis（2-ethylhexyl） phthalate （DEHP） in inducing cholestasis and liver injury in mice. MethodsIn the in vivo experiment， adult female ICR mice were randomly divided into control group （corn oil） and DEHP group （200 mg/kg/d）， and a model of cholestasis was established by intragastric administration for 4 weeks. After blood and liver tissue samples were collected from all mice， a biochemical analyzer was used to measure the level of total bile acid （TBA） in serum and the liver， and a microplate reader was used to measure alkaline phosphatase （ALP） and gamma-glutamyl transpeptidase （GGT）； HE staining was used to observe the pathological changes of the liver； RT-PCR was used to measure the mRNA expression levels of the inflammatory factors interleukin-1β （IL-1β）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α） in the liver； liquid chromatography/triple quadrupole mass spectrometry was used to measure the bile acid profile in the liver of mice. In the in vitro experiment， AML-12 mouse hepatocytes were cultured and treated with DEHP （250 µmol/L）， DCA （125 µmol/L）， and CDCA （125 µmol/L） for 24 hours， and RT-PCR was used to measure the mRNA expression levels of the inflammatory cytokines IL-1β， IL-6， and TNF-α. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the LSD-t test was used for further comparison between two groups. ResultsThe in vivo experiment showed that compared with the control group， the DEHP group had significant increases in the serum levels of TBA， ALP， and GGT and the level of TBA in the liver （the t values are respectively -4.396， -5.109， -8.504， -3.792 and -7.974， all P<0.05，）. Compared with the control group， the DEHP group had significant increases in cholic acid， chenodeoxycholic acid， taurocholic acid， deoxycholic acid， and ursodeoxycholic acid （the t values are respectively -2.802， -3.177， -2.633， -2.874 and -2.311， all P<0.05）. HE staining of the liver showed that the mice in the DEHP group had enlargement of the portal area， bile duct deformation， inflammatory cell infiltration around the bile duct， and significant increases in the mRNA expression levels of the inflammatory factors IL-1β， IL-6， and TNF-α in the liver （the t values are respectively -2.539， -2.823 and -4.636， all P<0.05）. The in vitro experiment showed that the actual difference in hepatocyte viability after 0-1 000 µmol/L DEHP treatment does not exceed 15%， but there were significant increases in the mRNA expression levels of the inflammatory cytokines IL-1β， IL-6， and TNF-α after treatment with DEHP at different concentrations of 125 µmol/L， 250 µmol/L， and 500 µmol/L （all P<0.05）. Compared with DEHP stimulation alone， the combined stimulation of CDCA and DEHP upregulates the cytokine in hepatocyte IL-1β mRNA levels （P<0.01）； the combined stimulation of DCA and DEHP can significantly increase the cytokine in hepatocyte IL-1β and IL-6 mRNA levels （all P<0.01）. ConclusionDEHP exposure can cause cholestasis and induce liver inflammation in mice， possibly by promoting the production of toxic bile acids and the secretion of inflammatory factors.

Diagnostic efficacy of AI in rib fracture under CT images with different reconstruction slice thickness / 重庆医学

Objective To investigate the diagnostic efficiency of artificial intelligence(AI)in rib frac-ture under the computed tomography(CT)images with different reconstruction slice thickness.Methods The first CT images of 100 patients with rib fractures were selected,and the interval-free recon-struction was carried out with the thickness of 0.625 mm,1.250 mm,2.500 mm and 5.000 mm,respectively.The rib fracture screening function of AI was used to automatically detect the CT images of four groups,and the diagnostic efficiency of AI for rib fracture under different reconstruction thickness conditions was com-pared.Results The sensitivity of AI in the diagnosis of rib fracture at 0.625 mm,1.250 mm,2.500 mm and 5.000 mm thickness was 99.32%(436/439),98.41%(432/439),89.52%(393/439)and 83.60%(367/439),respectively.The false positive rate was 4.80%(22/458),0.92%(4/436),0.76%(3/396)and 0.27%(1/368).The diagnostic sensitivity of AI in 0.625 mm and 1.250 mm thickness was higher than that in 2.500 mm and 5.000 mm,and the difference was statistically significant(P<0.05),while there was no significant difference in the thickness of 0.625 mm and 1.250 mm.The false positive rate of AI in the diagnosis of 0.625 mm slice thickness was higher than that of 1.250 mm,2.500 mm and 5.000 mm,and the difference was sta-tistically significant(P<0.05),while there was no significant difference in the thickness of 1.250 mm,2.500 mm and 5.000 mm(P>0.05).Conclusion The diagnostic efficiency of AI in 1.250 mm CT images is better than that in 0.625 mm,2.500 mm and 5.000 mm CT images.

Value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of Mycoplasma pneumoniae pneumonia in children / 国际检验医学杂志

Objective To analyze the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of Mycoplasma pneumoniae pneumonia(MPP)in children.Methods A total of 120 children with MPP who were admitted to the hospital from January 2020 to January 2023 were selected as the observation group,80 children with pulmonary tuberculosis(TB)were selected as the control group.During the same pe-riod,120 healthy children who were examined at the hospital check-up center were selected as the health group.The clinical data from each group were retrospective analyzed,and fasting venous blood from subjects was collected.The levels of T-lymphocyte subsets CD3+,CD4+and CD8+were detected by flow cytometry in each group,and the CD4+/CD8+was calculated.The levels of interferon(IFN)-y,interleukin-8(IL-8),inter-leukin-10(IL-10)and interleukin-13(IL-13)were measured by enzyme-linked immunosorbent assay,and the levels of peripheral blood T-lymphocyte subsets and cytokines in each group were compared.The receiver op-erating characteristic(ROC)curve was used to assess the value of peripheral blood T-lymphocyte subsets and cytokines in the diagnosis of MPP in children.Results The levels of CD3+,CD4+,CD4+/CD8+,IL-13,and IL-10 in the observation group and control group were significantly lower than those in the health group(P＜0.05),while CD8+,IL-8,and IFN-γ levels were significantly higher than those in the health group(P＜0.05).Compared with the control group,the observation group had higher levels of CD3+,CD8+,IL-8,IFN-γand lower levels of CD4+,CD4+/CD8+(P＜0.05),and there were no statistically significant differences in IL-8 and IL-13 levels between the two groups(P＞0.05).The ROC curve results showed that the area under the curve(AUC)or CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13,and IL-10 for the diagnosis of MPP in children were 0.751,0.687,0.784,0.864,0.798,0.672,0.650,and 0.811,and AUC of the combined detection was 0.924.Conclusion Children with TB and MPP have significantly decreased immune function in the early stages of the disease,and abnormal expression of peripheral blood T-lymphocyte subsets and cytokines.Com-pared with TB children,MPP children have lower levels of CD4+,CD4+/CD8+,and higher levels of CD3+,CD8+,IL-10 and IFN-γ,and the T-lymphocyte subsets and cytokine levels are closely related to the changes in the patients'condition.The combined detection of CD3+,CD4+,CD8+,CD4+/CD8+,IL-8,IFN-γ,IL-13 and IL-10 provides a theoretical basis for identifying and diagnosing early MPP in children.

Efficacy of NT-proBNP,hs-CRP,D-D,and PCT in predicting heart failure after acute myocardial infarction based on ROC and DCA curve analysis / 国际检验医学杂志

Objective To investigate the predictive efficacy of serum aminoterminal brain natriuretic pep-tide precursor(NT-proBNP),hypersensitive C-reactive protein(hs-CRP),D-dimer(D-D)and procalcitonin(PCT)in heart failure after acute myocardial infarction(AMI),Methods A total of 100 AMI patients admit-ted to the hospital from July 2021 to July 2023 were enrolled in the study as the observation group,In addi-tion,100 healthy people who underwent physical examination in the hospital during the same period were en-rolled as the control group,The serum levels of NT-proBNP,hs-CRP,D-D and PCT were detected and com-pared between the observation group and the control group,The AMI patients enrolled in the study were fur-ther divided into the heart failure group(31 cases)and the non-heart failure group(69 cases)according to the presence or absence of heart failure.The serum levels of NT-proBNP,hs-CRP,D-D,and PCT were compared between the two groups,Univariate analysis and multivariate Logistic regression analysis were used to analyze the risk factors of heart failure after AMI,Receiver operating characteristic(ROC)curve and decision curve a-nalysis(DCA)were used to analyze the predictive efficacy of serum NT-proBNP,hs-CRP,D-D and PCT for heart failure after AMI.Results The levels of serum NT-proBNP,hs-CRP,D-D and PCT in the observation group were higher than those in the control group(P＜0.05).The serum levels of NT-proBNP,hs-CRP,D-D and PCT in the complicated heart failure group were higher than those in the non-heart failure group(P＜0.05),Body mass index(BMI),smoking history,hypertension,number of diseased vessels,serum uric acid(SUA),low-density lipoprotein cholesterol(LDL-C),NT-proBNP,hs-CRP,D-D and PCT were risk factors for heart failure after AMI(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of combined detection of serum NT-proBNP,hs-CRP,D-D and PCT for predicting heart failure after AMI was 0.857(95％CI:0.811-0.948),the sensitivity was 96.12％,and the specificity was 91.28％,which were higher than the corresponding efficacy indexes of single detection(P＜0.05).DCA analysis showed that when the high-risk threshold was 0-0.99,the net benefit rate was greater than 0,which had clinical significance,When the threshold was 0-0.76,the net benefit rate of combined detection of serum NT-proBNP,hs-CRP,D-D and PCT was better than that of serum NT-proBNP,hs-CRP,D-D and PCT alone.Conclusion Combined detection of serum NT-proBNP,hs-CRP,D-D and PCT can improve the predictive efficiency of AMI compli-cated with heart failure,BMI,smoking history,hypertension,number of diseased vessels,SUA,LDL-C,NT-proBNP,hs-CRP,D-D and PCT are risk factors for AMI complicated with heart failure.

Determination of the Contents of Three Lignans in Dendrobium fimbriatum Hook / 广州中医药大学学报

Objective To establish the method for content determination of three lignans of Dendrobium Fimbriatum Hook..Methods The lignans in Dendrobium tasselii were identified by high-performance liquid chromatography/multi-stage mass spectrometry(HPLC-ESI/MSn)coupled with ultraviolet absorption spectrometry(UV)coupled with retention time localization of high-performance liquid chromatography(HPLC).The separation was carried out on a Kromasil 100-5 C18 column(4.6 mm×250 mm,5 μm)using a gradient elution of acetonitrile-0.1%formic acid solution as the mobile phase,the volume flow rate was 0.8 mL·min-1 and the column temperature was 35℃,and the mass spectrometry was performed using an ESI ion source with the data collected in the negative ion mode.The HPLC content was determined on the same column as that of MS analysis,with the mobile phase methanol + acetonitrile(V/V=1∶1)-0.01 mol/L ammonium acetate solution,gradient elution,flow rate of 0.8 mL·min-1,column temperature of 40℃,and detection wavelength of 215 nm.Results Syringaresinol di-O-glucoside and(-)-Syringaresinol 4-O-β-D-glucopyranoside and DL-Syringaresinol were identified from Dendrobium fimbriatum Hook.,and the results of content determination showed that the linear ranges of above three components were respectively 0.1701-3.4020,0.1020-2.0400,0.0403-0.8060 μg(r≥0.9995),the average recoveries were in the range of 97.71%-101.67%,and the relative standard deviations(RSDs)were all less than 3.0%.The contents of Syringaresinol di-O-glucoside and(-)-Syringaresinol 4-O-β-D-glucopyranoside and DL-Syringaresinol in the 10 batches of samples were 0.7779-1.3852,0.0734-0.1966,0.0295-0.1882 mg·g-1.Conclusion This research method can provide a reference basis for the quality evaluation method of Dendrobium fimbriatum Hook..

Di'ao Xinxuekang activates IRS-1/PI3K/Akt signal pathway to improve insulin resistance in nonalcoholic fatty hepatitis mice / 中国临床药理学与治疗学

AIM: To study the effect and mechanism of Di'ao Xinxuekang (DXXK) on insulin resistance in nonalcoholic steatohepatitis (NASH) mice. METHODS: C57BL/6J mice were randomly divided into normal group and model group. After 16 weeks of high-fat diet, the model group was randomly divided into model group and Pioglitazone group (6.0 mg · kg

Rutin forestalls dysregulated cardiac bioenergetics in bisphenol A and dibutyl phthalate-exposed rats through PPAR? and AMPK modulation

Objectives: Proper cardiac function is greatly dependent on adequate supply and metabolism of energy substrates. Environmental pollutants exposure including plasticizers can trigger adverse cardiac metabolic events. This study was designed to investigate the ameliorative effect of rutin (Rt) on dysregulated cardiac energy metabolism in plasticizer-exposed rats. Materials and Methods: Forty-two rats were randomised into seven groups (n = 6): Control (0.1% dimethyl sulfoxide), bisphenol A (BPA, 25 mg/kg, p.o), dibutyl phthalate (DBP, 25 mg/kg, p.o), BPA + Rt 25 mg/kg, Rt 50 mg/kg, DBP + Rt (25 mg/kg, Rt 50 mg/kg), BPA + DBP and BPA + DBP + Rt, daily for 21 days. Results: BPA and DBP exposure increased plasma glucose, reduced insulin, and increased plasma and cardiac free fatty-acid. Cardiac glucose-6-phosphate level, hexokinase and pyruvate dehydrogenase activities increased in DBP while BPA reduced these variables. Cardiac glucose transporter-4 expression was reduced in BPA group, while cardiac peroxisome proliferator-activated receptor-alpha (PPAR?) and AMP-activated protein kinase (AMPK) expression increased in BPA and DBP-treated rats. However, Rt administration prevents impaired cardiac bioenergetics and glucometabolic regulation. Conclusion: Summarily, Rt improves BPA and DBP-impaired cardiac bioenergetics through PPAR? and AMPK modulation.

Invitro Comparative Analysis of the Flexural Strength of 4 Different Commercially Available Provisional Materials Used in Fixed Partial Dentures – An Original Research

Background: Provisional Prosthesis in fixed partial dentures are subjected to Flexure under stress. Selection of appropriate material for fabrication of Provisional is of utmost importance as the Provisional prosthesis has to remain in function till definitive prosthesis is delivered. Material & Methods: Bar type specimens of four different commercially available brands for provisional restorations fabricated according to ADA specification No. 27 and immersed in artificial saliva. The specimens were fractured under 3-point loading test. Results: The flexural strength ranged between 60 to 110 Mpa. BisGMA Auto polymerizing composite resin from Dentsply Caulk shows the highest flexural strength. Conclusion: Within the limitations of this study, the flexural strengths were material specific rather than category one. The BisGMA composite based resin shows significantly higher flexural strength over other materials

Overview of Chinese medicinal materials industry in 2021 and development suggestions / 中国中药杂志

This paper introduced the overview of the "eight trends" of Chinese medicinal materials(CMM) industry in 2021, analyzed the problems of CMM production, and put forward development suggestions. Specifically, "eight trends" could be summarized as follows.(1) The growing area of CMM tended to be stable, and some provinces began to release the local catalog of Dao-di herbs.(2) The protection process of new varieties accelerated, and a number of excellent varieties were bred.(3) The theory of ecological cultivation was further enriched, and the demonstration effect of ecological cultivation technology was prominent.(4) Some CMM realized complete mechanization and formed typical model cases.(5) The number of cultivation bases using the traceability platform increased, and provincial internet trading platforms were set up.(6) The construction of CMM industrial clusters accelerated, and the number of provincial-level regional brands increased rapidly.(7) Many new agricultural business entities were founded nationwide, and a variety of methods were used to drive the intensified development of CMM.(8) A number of local TCM laws were promulgated, and the management regulation of food and medicine homology substances catalogs was issued. On this basis, four suggestions for CMM production were proposed.(1) It is suggested to speed up the formulation of the national catalog of Dao-di herbs and carry out the certification of Dao-di herbs production bases.(2) Ecological planting of forest and grassland medicine should be further strengthened in terms of technical research and promotion based on the principle of ecological priority.(3) The basic work of disaster prevention should be paid more attention and technical measures for disaster mitigation should be developed.(4) The planted area of commonly used CMM should be incorporated into the national regular statistical system.

Specific DNA barcodes, germplasm resources, and genetic diversity of Eleutherococcus senticosus / 中国中药杂志

Eleutherococcus senticosus is one of the Dao-di herbs in northeast China. In this study, the chloroplast genomes of three E. senticosus samples from different genuine producing areas were sequenced and then used for the screening of specific DNA barcodes. The germplasm resources and genetic diversity of E. senticosus were analyzed basing on the specific DNA barcodes. The chloroplast genomes of E. senticosus from different genuine producing areas showed the total length of 156 779-156 781 bp and a typical tetrad structure. Each of the chloroplast genomes carried 132 genes, including 87 protein-coding genes, 37 tRNAs, and 8 rRNAs. The chloroplast genomes were relatively conserved. Sequence analysis of the three chloroplast genomes indicated that atpI, ndhA, ycf1, atpB-rbcL, ndhF-rpl32, petA-psbJ, psbM-psbD, and rps16-psbK can be used as specific DNA barcodes of E. senticosus. In this study, we selected atpI and atpB-rbcL which were 700-800 bp and easy to be amplified for the identification of 184 E. senticosus samples from 13 genuine producing areas. The results demonstrated that 9 and 10 genotypes were identified based on atpI and atpB-rbcL sequences, respectively. Furthermore, the two barcodes identified 23 genotypes which were named H1-H23. The haplotype with the highest proportion and widest distribution was H10, followed by H2. The haplotype diversity and nucleotide diversity were 0.94 and 1.82×10~(-3), respectively, suggesting the high genetic diversity of E. senticosus. The results of the median-joining network analysis showed that the 23 genotypes could be classified into 4 categories. H2 was the oldest haplotype, and it served as the center of the network characterized by starlike radiation, which suggested that population expansion of E. senticosus occurred in the genuine producing areas. This study lays a foundation for the research on the genetic quality and chloroplast genetic engineering of E. senticosus and further research on the genetic mechanism of its population, providing new ideas for studying the genetic evolution of E. senticosus.

Characteristics, origin, and processing of Poria in Qing Dynasty Palace:evidence of both historical relics and documents / 中国中药杂志

Poria(Fu Ling) is a bulk traditional Chinese medicine(TCM)with a long history and complex varieties. The royal medical records of the Qing Dynasty include multiple medicinal materials of Fu Ling, such as Bai Fu Ling(white Poria), Chi Fu Ling(rubra Poria), and Zhu Fu Ling(Poria processed with cinnabaris). The Palace Museum preserves 6 kinds of specimens including Fu Ling Ge(dried Poria), Bai Fu Ling, Chi Fu Ling, Zhu Fu Ling, Bai Fu Shen(white Poria cum Radix Pini), and Fu Shen Mu(Poria cum Radix Pini). After trait identification and textual research, we found that Fu Ling Ge was an intact sclerotium, which was processed into Fu Ling Pi(Poriae Cutis), Bai Fu Ling and other medicinal materials in the Palace. The Fu Ling in the Qing Dynasty Pa-lace was mainly from the tribute paid of the officials in Yunnan-Guizhou region. The tribute situation was stable in the whole Qing Dynasty, and changed in the late Qing Dynasty. The cultural relics of Fu Ling in the Qing Dynasty Palace confirm with the archival documents such as the royal medical records and herbal medicine books, providing precious historical materials for understanding Fu Ling in the Qing Dynasty and a basis for the restoration of the processing of the Fu Ling in the Qing Dynasty Palace.

Content determination of seven active components of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats by UPLC-MS/MS / 中国中药杂志

In the present study, the contents of seven active components [genipinic acid(GA), protocatechuic acid(PCA), neochlorogenic acid(NCA), chlorogenic acid(CA), cryptochlorogenic acid(CCA),(+)-pinoresinol di-O-β-D-glucopyranosid(PDG), and(+)-pinoresinol 4'-O-β-D-glucopyranoside(PG)] of Eucommiae Cortex in aortic vascular endothelial cells of spontaneously hypertensive rats(SHR) were simultaneously determined by ultra-high liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS). The qualified SHR models were selected. The primary aortic endothelial cells(VECs) of rats were separated and cultured by ligation and adherence, followed by subculture. After successful identification, an UPLC-MS/MS method for simultaneously determining the contents of GA, PCA, NCA, CA, CCA, PDG, PG in seven components of Eucommiae Cortex in VECs was established, including specificity, linearity, matrix effect, recovery, accuracy, precision and stability. The established method had the lo-west limit of quantification of 0.97-4.95 μg·L~(-1), accuracy of 87.26%-109.6%, extraction recovery of 89.23%-105.3%, matrix effect of 85.86%-106.2%, and stability of 86.00%-112.5%. Therefore, the established accurate UPLC-MS/MS method could rapidly and simultaneously determine the contents of the seven active components of Eucommiae Cortex in VECs of SHRs, which provided a refe-rence for the study of cellular pharmacokinetics of active components of Eucommiae Cortex extract.

Leptospiral c-di-AMP-binding protein regulates potassium ion transport / 中华微生物学和免疫学杂志

Objective:To investigate whether KtrA was a binding protein of c-di-AMP, the second messenger in Leptospira, and to explore the function and regulatory mechanism of the c-di-AMP-KtrA/B system. Methods:KtrA gene was amplified by PCR and cloned into pET42a plasmid to construct the pET42a ktrA prokaryotic expression vector. Then the vector was transferred into E. coli BL21DE3 to construct an engineering bacterium E. coli BL21DE3 pET42a-ktrA for the expression of recombinant KtrA (rKtrA). The expressed rKtrA was purified by affinity chromatography. BIAcore technology was used to detect the binding ability of rKtrA to c-di-AMP. Bacterial two-hybrid analysis was used to analyze the interaction between KtrA and KtrB in the leptospiral Ktr system with or without exogenetic c-di-AMP. The above genes were then complemented into the potassium transport-deficient E. coli mutants to analyze the function of the c-di-AMP-KtrA/B pathway. Results:An prokaryotic engineering bacterium for the expression of ktrA gene of Leptospira was constructed successfully. The purified rKtrA could specifically bind to c-di-AMP. There was interaction between KtrA and KtrB, but the interaction could be dissociated by c-di-AMP. The KtrA/B system was involved in potassium ion uptake and it was negatively regulated by c-di-AMP. Conclusions:Leptospiral KtrA was a c-di-AMP-binding protein and the c-di-AMP-KtrA/B system was involved in potassium ion transport.

Study on the mechanism of Dihuang Decoction in treating Alzheimer disease and diabetes mellitus with same method based on network pharmacology / 国际中医中药杂志

Objective:To study the common pharmacodynamic substance basis and potential mechanism of Dihuang Decoction in treating Alzheimer disease (AD) and diabetes mellitus (DM) with same method based on network pharmacology; To provide bioinformatics basis.Methods:The effective components of Dihuang Decoction were retrieved through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and CNKI. The drug targets were obtained by combining and UniProt database. The targets of AD and DM related diseases were obtained by GeneCards, OMIM and TTD databases respectively. Cytoscape 3.7.1 software was used to construct the Disease - drug - component - target network. R studio software was used to construct a Circos diagram of the effective compounds and disease targets.Protein-protein interaction (PPI) network was constructed using STRING platform. GO enrichment and KEGG enrichment analysis was conducted through DAVID database, Metascape, R Studio software.Results:A total of 206 active components were obtained; PPI network construction screened 51 key targets; GO enrichment analysis revealed the functions of GABA, cholinergic synapse, estrogen response, BCL-2 family protein complex and so on; KEGG enrichment analysis revealed FoxO signaling pathway, HIF-1 signaling pathway, insulin resistance pathway and other pathways.Conclusion:Dihuang Decoction has the synergistic characteristics of multiple components, multiple targets and multiple pathways in treating AD and DM with same method, mainly through proanthocyanidin B7, proanthocyanidin B5, proanthocyanidin B1 and other active ingredients, acting on TNF, IL-6, ESR1, PPARG, AKT1 and other targets, regulating FoxO signaling pathway, HIF-1 signaling pathway, etc.

Value of serum IgG4 and IgG4/IgG ratio in differential diagnosis of autoimmune diseases / 中国基层医药

Objective:To investigate the value of immunoglobulin G4 (IgG4) and IgG4/ immunoglobulin G (IgG) ratio in the differential diagnosis of IgG4-related diseases (IgG4-RD) and other autoimmune diseases.Methods:A total of 35 patients with IgG4-RD and 937 patients with autoimmune diseases who received treatment in Beijing Hospital from January 2021 to July 2022, and 200 subjects who concurrently underwent health checkups in the same hospital were included in this study. The IMMAGE 800 and BN II automatic special protein analyzers were used to detect IgG and IgG4. The receiver operating characteristic (ROC) curve of IgG4 and IgG4/IgG ratio was plotted.Results:Serum IgG4 level and IgG4/IgG ratio in the IgG4-RD group were 2.83 (2.01, 5.07) g/L and 25% (18%, 43%) respectively, which were higher than 0.35 (0.16, 0.72) g/L, 3% (1%, 6%) in the autoimmune disease group and 0.27 (0.14, 0.49) g/L, 2% (1%, 4%) in the healthy control group ( U = 795.50, 82.50, 1 744.50, 205.50, all P < 0.001). Taking IgG4 ≥ 1.35 g/L as the standard, patients with IgG4 ≥ 1.35 g/L in the three groups were screened out. There was a statistically significant difference in IgG4/IgG ratio between the IgG4-RD group and the non-IgG4-RD group ( U = 453.50, P < 0.001). The ROC curve of IgG4 and IgG4/IgG ratio showed that when IgG4 was 1.47 g/L, the sensitivity was 91.7%, the specificity was 83.5%, and the area under the ROC curve was 0.96. When IgG4/IgG was 12.5%, the sensitivity was 91.4%, the specificity was 85%, and the area under the ROC curve was 0.96. Taking IgG4 ≥ 1.47 g/L and IgG4/IgG ≥ 12.5% as the diagnostic criteria of IgG4-RD, the sensitivity was 94.3%, the specificity was 85.9%, and the area under the ROC curve was 0.96, which were higher than the sensitivity (87.2%) and diagnostic specificity (82.6%) provided by IgG4 alone. Conclusion:Because non-IgG4-RD diseases can also have the phenomenon of increased IgG4, when IgG4 ≥ 1.47 g/L is taken as the diagnostic criteria, its diagnostic sensitivity and specificity are the highest. Combined detection of IgG4 and IgG4/IgG ratio can increase the diagnostic efficacy of IgG4-RD.