Reappraisals of biological behaviors of PDGFRA mutant gastrointestinal stromal tumor / 中华病理学杂志

Objective: To investigate the biological behavior spectrum of platelet-derived growth factor alpha receptor (PDGFRA)-mutant gastrointestinal stromal tumor (GIST), and to compare the clinical values of the Zhongshan method of benign and malignant evaluation with the modified National Institutes of Health (NIH) risk stratification. Methods: A total of 119 cases of GIST with PDGFRA mutation who underwent surgical resection at Zhongshan Hospital, Fudan University from 2009 to 2020 were collected. The clinicopathological data, follow-up records, and subsequent treatment were reviewed and analyzed statistically. Results: There were 79 males and 40 females. The patients ranged in age from 25 to 80 years, with a median age of 60 years. Among them, 115 patients were followed up for 1-154 months, and 13 patients progressed to disease. The 5-year disease-free survival (DFS) and overall survival (OS) were 90.1% and 94.1%, respectively. According to the modified NIH risk stratification, 8 cases, 32 cases, 38 cases, and 35 cases were very-low risk, low risk, intermediate risk, and high risk, and 5-year DFS were 100.0%, 95.6%, 94.3%, and 80.5%, respectively. There was no significant difference in prognosis among the non-high risk groups, only the difference between high risk and non-high risk groups was significant (P=0.029). However, the 5-year OS was 100.0%, 100.0%, 95.0% and 89.0%, and there was no difference (P=0.221). According to the benign and malignant evaluation Zhongshan method, 43 cases were non-malignant (37.4%), 56 cases were low-grade malignant (48.7%), 9 cases were moderately malignant (7.8%), and 7 cases were highly malignant (6.1%). The 5-year DFS were 100.0%, 91.7%, 77.8%, 38.1%, and the difference was significant (P<0.001). The 5-year OS were 100.0%, 97.5%, 77.8%, 66.7%, the difference was significant (P<0.001). Conclusions: GIST with PDGFRA gene mutation shows a broad range of biological behavior, ranging from benign to highly malignant. According to the Zhongshan method, non-malignant and low-grade malignant tumors are common, the prognosis after surgery is good, while the fewer medium-high malignant tumors showed poor prognosis after surgical resection. The overall biological behavior of this type of GIST is relatively inert, which is due to the low proportion of medium-high malignant GIST. The modified NIH risk stratification may not be effective in risk stratification for PDGFRA mutant GIST.

Association between ELL2 polymorphism and susceptibility to pleomorphic adenoma of salivary gland / 解放军医学杂志

Objective To analyze the correlation between the ELL2 gene 1119 T>C polymorphism and the susceptibility to pleomorphic adenoma of the salivary gland.Methods The pedigree of the pleomorphic adenoma family of salivary gland was drawn.The exons of ELL2 gene in 5 members of salivary pleomorphic adenoma family were sequenced.A case-control study was conducted.One hundred and twelve patients with pleomorphic adenoma of the salivary gland who visited the Department of Oral and Maxillofacial Surgery of Shanxi Bethune Hospital from January 2016 to July 2020 were taken as case group,and 176 healthy examinees from January 2019 to January 2020 were taken as control group with age and sex as matching conditions.The 1119 T>C polymorphism of ELL2 genes in the two groups were detected with high resolution melting(HRM)curve.Chi-square test was adopted to analyze the correlation between gene polymorphism and the occurrence of pleomorphic adenoma of the salivary gland,stratified analysis was performed to evaluate the synergistic effect of smoking and genotype,and real time quantitative reverse transcription polymerase chain reaction(RT-PCR)was used to detect the expression level of ELL2 in individuals with different genotypes.Results The 1119 T>C polymorphism site existed in the exon 8 of ELL2 gene in a family with pleomorphic adenoma of salivary gland.The results of case-control study showed that the genotype frequency of homozygous CC was significantly higher in patients with pleomorphic adenoma of salivary gland than that in the controls(24.1%vs.11.9%,P=0.002).Homozygous CC was associated with increased risk for developing pleomorphic adenoma of salivary gland(OR=3.059,95%CI 1.494-6.263)in this cohort.Stratification analysis showed that smoking and 1119C allele cooperated to increase the risk of pleomorphic adenoma of salivary gland(OR=3.200,95%CI 1.460-7.014).The expression level of ELL2 mRNA in CC genotype was significantly higher than that in individuals with CT or TT genotype(P<0.05).Conclusion The genetic variation of ELL2 may play an important role in the occurrence of pleomorphic adenoma of salivary gland,and smoking combined with the 1119C allele increased the risk of this disease.

Effect of fumigation therapy of Flos Farfarae in cigarette smoke-induced lung injury mice based on metabolomics / 药学学报

This study aimed to investigate the effect of Flos Farfarae (FF) fumigation on cigarette smoke-induced lung injury mice, and analyze the metabolic profile of lung tissue by metabolomics. All animal experiments were conducted under the guidance and approval of the Animal Ethics Review Committee of Shanxi University (Approval number: SXULL2019014). By using HS-GC-MS to analyze volatile components of Flos Farfarae, 23 compounds were identified. The results showed that FF fumigation improved the lung tissue morphology of cigarette smoke-induced lung injury mice, lowered the levels of interleukin 6 (IL-6) and interleukin-1β (IL-1β). The lung tissue samples were applied for metabolomic analysis based on UHPLC-QTOF MS, the results showed that 70 metabolites were changed in the lung tissue of mice after cigarette exposure, and 35 of them could be regulated, including lysophosphatidylcholine (LPC), 12-HETE, adenosine, and xanthine. These metabolites, such as LPC, 12-HETE, adenosine, and xanthine were mainly associated with the body's inflammatory response. It was observed that these metabolites are primarily involved in purine metabolism, arachidonic acid metabolism, phospholipid metabolism, and pyruvate metabolism pathways. These findings suggest that the volatile terpenoids in the FF may regulate the metabolites associated with the inflammatory response in the lung tissue, such as lysophosphatidylcholine, 12-HETE, and adenosine, which could further alleviate lung inflammation induced by cigarette smoke through the metabolic pathways of purine metabolism and others. This study proved the scientific basis of the traditional application of FF fumigation, and provided a theoretical basis for the further product development.

Effect of Shugan Tiaoshen acupuncture combined with western medication on depression-insomnia comorbidity due to COVID-19 quarantine: a multi-central randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the effect of Shugan Tiaoshen acupuncture (acupuncture for soothing the liver and regulating the mentality) combined with western medication on depression and sleep quality in the patients with depression-insomnia comorbidity due to COVID-19 quarantine, and investigate the potential mechanism from the perspective of cortical excitability.@*METHODS@#Sixty patients with depression-insomnia comorbidity due to COVID-19 quarantine were randomly divided into an acupuncture group and a sham-acupuncture group, 30 cases in each one. The patients of both groups were treated with oral administration of sertraline hydrochloride tablets. In the acupuncture group, Shugan Tiaoshen acupuncture was supplemented. Body acupuncture was applied to Yintang (GV 24+), Baihui (GV 20), Hegu (LI 4), Zhaohai (KI 6), Qihai (CV 6), etc. The intradermal needling was used at Xin (CO15), Gan (CO12) and Shen (CO10). In the sham-acupuncture group, the sham-acupuncture was given at the same points as the acupuncture group. The compensatory treatment was provided at the end of follow-up for the patients in the sham-acupuncture group. In both groups, the treatment was given once every two days, 3 times a week, for consecutive 8 weeks. The self-rating depression scale (SDS) and insomnia severity index (ISI) scores were compared between the two groups before and after treatment and 1 month after the end of treatment (follow-up) separately. The cortical excitability indexes (resting motor threshold [rMT], motor evoked potential amplitude [MEP-A], cortical resting period [CSP]) and the level of serum 5-hydroxytryptamine (5-HT) were measured before and after treatment in the two groups.@*RESULTS@#After treatment and in follow-up, SDS and ISI scores were decreased in both groups compared with those before treatment (P<0.05), and the scores in the acupuncture group were lower than those in the sham-acupuncture group (P<0.05), and the decrease range in the acupuncture group after treatment was larger than that in the sham-acupuncture group (P<0.05). After treatment, rMT was reduced (P<0.05), while MEP-A and CSP were increased (P<0.05) in the acupuncture group compared with that before treatment. The levels of serum 5-HT in both groups were increased compared with those before treatment (P<0.05). The rMT in the acupuncture group was lower than that in the sham-acupuncture group, while MEP-A and CSP, as well as the level of serum 5-HT were higher in the acupuncture group in comparison with the sham-acupuncture group (P<0.05).@*CONCLUSION@#Shugan Tiaoshen acupuncture combined with western medication can relieve depression and improve sleep quality in the patients with depression-insomnia comorbidity due to COVID-19 quarantine, which is probably related to rectifying the imbalanced excitatory and inhibitory neuronal functions.

Domestic and international research status and trend of adolescent myopia: A bibliometrics based analysis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the current research progress, hot spots and future development trends of myopia in children and adolescents at home and abroad, thus providing references for the further research on the field.METHODS: Using China National Knowledge Infrastructure(CNKI)and Web of Science(WOS)as data sources, the literature on myopia in children and adolescents from January 1, 2003 to December 31, 2022 was collected. Visual analyses were conducted based on the quantity of the published articles, authors, publishing institutions, journals as well as keyword co-occurrence, clustering, timeline graph and emergence by using the VOSviewer and CiteSpace.RESULT: The number of publications on myopia research has increased steadily both articles in Chinese and English. Xian-Gui He and Saw, Seang-Mei have published the most papers, and the Chinese Journal of School Health and Investigative Ophthalmology &#x0026; Visual Science published the most related articles. Shanghai Eye Disease Prevention and Treatment Center, and Sun Yat-sen University are the institutions with the most publications in the area. Clinical observation is the main focus of research both domestically and internationally, with orthokeratology, outdoor activities, axial length, prevalence, and influencing factors attracting much attention.CONCLUSION: Current research on myopia in children and adolescents focuses on the epidemiology, intervention measures, and biological parameters of the condition. Future research on myopia epidemiology is likely to become a major focus of research in this area.

Component identification and analysis in vivo of Sanhan Huashi formula / 中国中药杂志

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

Research progress on multi-target regulation strategies of tumor microenvironment based on nano-drug delivery system / 药学学报

Tumor microenvironment (TME) is composed of endothelial cells, pericytes, immune cells, cancer-associated fibroblasts (CAFs), cancer stem cells (CSCs), extracellular matrix (ECM) and other components of the complex biological environment. TME interacts with the tumor cells through a large amount of signaling pathways, participates in the process of tumor progression, invasion, and metastasis. Hence, TME has become a potential therapeutic target for cancer treatment, exhibiting excellent therapeutic potential and research value in the field of cancer treatment. Currently, the novel nanotechnology has been widely applied in anticancer therapy, and nanotechnology-mediated drug delivery system is being explored to apply in TME modulation to inhibit tumor progression. Nanotechnology-mediated drug delivery has many advantages over traditional therapeutic modalities, including longer circulation times, improved bioavailability, and reduced toxicity. This review summarized the research of targeted nano-drug delivery based on TME regulation, including regulation strategies based on CSCs, CAFs, immune cells, ECM, tumor vascularization, exosomes, and microbiota. In addition, we summarized the advantages, opportunities, and challenges of TME regulation strategy compared with traditional treatment strategy, which provides a reference for the application of nano-drug delivery system based on TME regulation strategy in tumor precision therapy.

Component profile analysis of Sanhan Huashi Formula based on UHPLC-LTQ-Orbitrap-MS, GC-MS, and UHPLC-QQQ-MS/MS / 中国中药杂志

This study comprehensively analyzed the active components of Sanhan Huashi Formula using qualitative and quantitative mass spectrometry techniques, laying the foundation for understanding its pharmacological substance basis. UHPLC-LTQ-Orbitrap-MS and GC-MS technologies were used to analyze and identify the volatile and non-volatile components in Sanhan Huashi Formula. UHPLC-QQQ-MS/MS technology was used to simultaneously determine the content of 27 major active components in the formula. The results showed that 308 major chemical components were identified in Sanhan Huashi Formula, among which 60 compounds were identified by comparing with reference standards, mainly including alkaloids, flavonoids, coumarins, triterpenoid saponins, amino acids, and nucleosides. GC-MS technology preliminarily identified 52 volatile compounds, with γ-eudesmol and β-eudesmol as the main components. The quantitative results demonstrated good linearity(r>0.99) for the 27 active components, indicating the stability, simplicity, and reliability of the established method. Among them, amygdalin, nodakenin, arecoline, ephedrine, and pseudoephedrine had relatively high content and were presumably the main pharmacologically active substances. In conclusion, this study systematically and comprehensively characterized the major chemical components and patterns in Sanhan Huashi Formula, providing a basis for understanding its pharmacological mechanisms and clinical applications.

Establishment and clinical validation of an artificial intelligence YOLOv51 model for the detection of precancerous lesions and superficial esophageal cancer in endoscopic procedure / 中华肿瘤杂志

Objective: To construct the diagnostic model of superficial esophageal squamous cell carcinoma (ESCC) and precancerous lesions in endoscopic images based on the YOLOv5l model by using deep learning method of artificial intelligence to improve the diagnosis of early ESCC and precancerous lesions under endoscopy. Methods: 13, 009 endoscopic esophageal images of white light imaging (WLI), narrow band imaging (NBI) and lugol chromoendoscopy (LCE) were collected from June 2019 to July 2021 from 1, 126 patients at the Cancer Hospital, Chinese Academy of Medical Sciences, including low-grade intraepithelial neoplasia, high-grade intraepithelial neoplasia, ESCC limited to the mucosal layer, benign esophageal lesions and normal esophagus. By computerized random function method, the images were divided into a training set (11, 547 images from 1, 025 patients) and a validation set (1, 462 images from 101 patients). The YOLOv5l model was trained and constructed with the training set, and the model was validated with the validation set, while the validation set was diagnosed by two senior and two junior endoscopists, respectively, to compare the diagnostic results of YOLOv5l model and those of the endoscopists. Results: In the validation set, the accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the YOLOv5l model in diagnosing early ESCC and precancerous lesions in the WLI, NBI and LCE modes were 96.9%, 87.9%, 98.3%, 88.8%, 98.1%, and 98.6%, 89.3%, 99.5%, 94.4%, 98.2%, and 93.0%, 77.5%, 98.0%, 92.6%, 93.1%, respectively. The accuracy in the NBI model was higher than that in the WLI model (P<0.05) and lower than that in the LCE model (P<0.05). The diagnostic accuracies of YOLOv5l model in the WLI, NBI and LCE modes for the early ESCC and precancerous lesions were similar to those of the 2 senior endoscopists (96.9%, 98.8%, 94.3%, and 97.5%, 99.6%, 91.9%, respectively; P>0.05), but significantly higher than those of the 2 junior endoscopists (84.7%, 92.9%, 81.6% and 88.3%, 91.9%, 81.2%, respectively; P<0.05). Conclusion: The constructed YOLOv5l model has high accuracy in diagnosing early ESCC and precancerous lesions in endoscopic WLI, NBI and LCE modes, which can assist junior endoscopists to improve diagnosis and reduce missed diagnoses.

CO2 laser moxibustion for endometriosis related pelvic pain of cold coagulation and blood stasis: a randomized controlled trial / 中国针灸

OBJECTIVE@#To observe the clinical therapeutic effect of CO2 laser moxibustion on endometriosis related pelvic pain of cold coagulation and blood stasis.@*METHODS@#A total of 76 patients with endometriosis related pelvic pain of cold coagulation and blood stasis were randomized into a laser moxibustion group and a sham laser moxibustion group, 38 cases in each group. In the laser moxibustion group, moxibustion was applied at bilateral Zigong (EX-CA 1) using CO2 laser moxibustion instrument. In the sham laser moxibustion group, the manipulation of moxibustion was same as the laser moxibustion group, without laser output. The treatment was given once every other day, 30 min each time, 3 times a week for 4 weeks in both groups. Before and after treatment and follow-up of 3 months after treatment, the scores of Gracely box scale (GBS) and visual analogue scale (VAS) were observed, the usage of non-steroidal anti-inflammatory drug for the duration of the treatment and the average days of taking drugs were recorded in both groups.@*RESULTS@#Compared before treatment, the GBS and VAS scores were decreased after treatment and during follow-up in the laser moxibustion group (P<0.05), while those in the sham moxibustion group had no significant differences (P>0.05). Compared with the sham moxibustion group, the GBS and VAS scores were decreased after treatment and during follow-up (P<0.05), the cases and average days of taking drugs were less in the laser moxibustion group (P<0.05).@*CONCLUSION@#CO2 laser moxibustion can improve the pain symptom in patients with endometriosis related pelvic pain of cold coagulation and blood stasis, and reduce the use of analgesic drugs.

A preliminary study on the origin of neonatal intestinal microbiota during cesarean section / 中国实用妇科与产科杂志

OBJECTIVE: To explore the composition of microbial communities in vagina and intestine of the mother,the placenta and the neonatal meconium after cesarean section and to analyze the origin of neonatal intestinal microbiota.METHODS: Samples of intestine,vagina and placenta and neonatal meconium from 4 women who underwent cesarean section in Yan'an Hospital of Kunming City in October 2016 were collected.The high-throughput sequencing technology was used to sequence the 16 S rRNA gene,and the composition of the microbial communities and the relationship among the samples were analyzed.RESULTS: Firstly,comparing the number of OTUs in different samples,it was found that the number in placenta was the highest,followed by the intestine and vagina,and the least was in the neonatal meconium.Secondly,each sample was analyzed by PCA clustering,and it was found that the neonatal meconium was affected least by individual differences but the vagina was affected most.The intestinal and placental microbial communities had certain similarities.Finally,comparing the microbial community composition of each sample,it was found that the highest abundance in the neonatal meconium and placenta was Proteobacteria,and in the intestine and vagina it was the Firmicutes.CONCLUSION: During the caesarean section,maternal microbiota transfer may be from the mother's intestine to the placenta and then to the infant's intestine.

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics / 中国实验方剂学杂志

Objective: To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS). Method: CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software. Result: Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione. Conclusion: Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.

Increased Endogenous Sulfur Dioxide Involved in the Pathogenesis of Postural Tachycardia Syndrome in Children: A Case-Control Study / 中华医学杂志(英文版)

<p><b>Background</b>The pathogenesis of postural tachycardia syndrome (POTS) remains unclear. This study aimed to explore the changes and significance of sulfur dioxide (SO) in patients with POTS.</p><p><b>Methods</b>The study included 31 children with POTS and 27 healthy children from Peking University First Hospital between December 2013 and October 2015. A detailed medical history, physical examination results, and demographic characteristics were collected. Hemodynamics was recorded and the plasma SOwas determined.</p><p><b>Results</b>The plasma SOwas significantly higher in POTS children compared to healthy children (64.0 ± 20.8 μmol/L vs. 27.2 ± 9.6 μmol/L, respectively, P < 0.05). The symptom scores in POTS were positively correlated with plasma SOlevels (r = 0.398, P < 0.05). In all the study participants, the maximum heart rate (HR) was positively correlated with plasma levels of SO(r = 0.679, P < 0.01). The change in systolic blood pressure from the supine to upright (ΔSBP) in POTS group was smaller than that in the control group (P < 0.05). The ΔSBP was negatively correlated with baseline plasma SOlevels in all participants (r = -0.28, P < 0.05). In the control group, ΔSBP was positively correlated with the plasma levels of SO(r = 0.487, P < 0.01). The change in HR from the supine to upright in POTS was obvious compared to that of the control group. The area under curve was 0.967 (95% confidence interval: 0.928-1.000), and the cutoff value of plasma SOlevel >38.17 μmol/L yielded a sensitivity of 90.3% and a specificity of 92.6% for predicting the diagnosis of POTS.</p><p><b>Conclusions</b>Increased endogenous SOlevels might be involved in the pathogenesis of POTS.</p>

Fabrication of Walking Function Orthosis for Patients with Leg Length Discrepancy / 中国康复理论与实践

Objective To study the design and fabricating method of orthosis for patients with leg length discrepancy. Methods Ischial weight bearing orthosis with prosthetic feet was made through efforts from weight,beauty and durabili-ty.The fabrication process included taking a negative plaster cast,modifying the positive model,forming,align-ment,and fitting the device to the patients. Results The orthosis was compensated for the patients'height with good appearance and convenience wearing. Conclusion Ischial weight bearing orthosis with prosthetic feet can help patients reconstruct walking function,therefore can be recommended.

Effects of HDAC Inhibitor Scriptaid on IM9 Cell Line and Its Mechanism / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effect of HDAC inhibitor Scriptaid on multiple myeloma IM9 cells and preliminarily clarify the mechanism of Scriptaid-induced cell apoptosis.</p><p><b>METHODS</b>The cell viability, cell cycle and cell apoptosis were measured by CCK8 assay and flow cytometry respectively, the relative target gene expression levels were detected by RT-PCR, the effect of Scriptaid on p21 promoter activity was detected by using luciferase reporter assay.</p><p><b>RESULTS</b>Scriptaid inhibited IM9 cell viability in a dose-dependent manner. Scriptaid induced IM9 cell cycle arrest at G/M phase in a dose-dependent manner. Scriptaid triggered IM9 cell apoptosis was obviously, the mRNA levels of apoptosis-related proteins Caspase 9, Caspase 3 and PARP1 were also activated. The apoptosis-associated factors BAD, PTEN and p21 increased following treatment with different dose of Scriptaid, meanwhile, p21 promoter activity was also activated significantly.</p><p><b>CONCLUSION</b>HDAC inhibitor Scriptaid can promote IM9 cell apoptosis by transcriptional activation of p21 promoter in concentration-dependent manner.</p>

Effect of Interfering the ADAM10 by Lentivirul Vector-Mediated shRNA on Multiple Myeloma MM.1S Cell Proliferation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of interfering ADAM10 on proliferation and apoptosis of multiple myeloma MM.1S cells, and its possible mechanism.</p><p><b>METHODS</b>Four pairs of shRNA-coding sequences directed against different sites of ADAM10 mRNA were designed and inserted into lentiviral vector plasimd pLVshRNA-EGFP(2A) Puro for constructing the sh/ADAM10-1, sh/ADAM10-2, sh/ADAM10-3, sh/ADAM10-4 and sh/Con. These plasmids and lentiviral packaging plasmids were co-transfected into the packaging cells 293FT, then the virus particles were collected and the viral titer was assayed after concentration, and these viral particles were transfected to MM.1S cells. The flow cytometry was used to sort GFPcells. Real-time quantitative PCR, and Western blot were used to detect the effect of interfering the ADAM10 gene by lentiviral vector mediated shRNA. The proliferation-inhibition curve was plotted by CCK-8 method, the cell viability and apoptosis were detected by flow cytometry with Annexin V and 7-AAD staining, the transcripts of pro-apoptosis gene BAD, BAK, BIK, anti-apoptotic genes BCL-2, c-Myc and Notch1 target gene Hes-1 were detected by real-time PCR.</p><p><b>RESULTS</b>Lentivirus vector was successfully constructed, that could specifically interfere ADAM10 expression. Interfering ADAM10 gene could inhibit the MM.1S cell proliferation and induce apoptosis. After the interferencing ADAM10 gene the mRNA levels of pro-apoptosis gene BAD, BAK and BIK were increased, and the mRNA levels of anti-apoptotic genes BCL-2 and c-Myc were reduced. Q-PCR results showed that the mRNA level of Notch1 were increased, but that of Hes-1 were reduced.</p><p><b>CONCLUSION</b>Down-regulated ADAM10 expression can significantly inhibit multiple myeloma MM.1S cell proliferation and promote the apotosis. Its mechanism may be related to Notch1 signaling pathways.</p>

Effects of Different Sample Preparation Methods on Total Flavonoids and Puerarin Content from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province / 中国中医药信息杂志

Objective To establish UV spectrophotometry and HPLC methods for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix and Puerariae Thomsonii Radix; To compare the ultrasonic method at room temperature, conventional refluxing method and ultrasonic method at heating conditions at the aspect of content determinations. Methods The content determinations of total flavonoids was determined by UV spectrophotometry at 250 nm; the content of puerarin was determined by HPLC with octadecylsilane-bonded silica gel as the stationary phase, a mixture of methanol and water (25:75) as the mobile phase, 256 nm as the detection wavelength, 1.0 mL/min as the flow rate. Results Contents of total flavonoids in Puerariae Lobatae Radix by ultrasonic method at room temperature, conventional refluxing method and ultrasonic method were15.09%, 14.48%, and 12.71% (n=3), respectively. The contents of puerarin were 4.37%, 4.09%, and 3.80% (n=3), respectively. Contents of total flavonoids in Puerariae Thomsonii Radix were 2.09%, 2.23%, and 2.17% (n=3), respectively. The contents of puerarin were 0.50%, 0.53%, and 0.52% (n=3), respectively. Conclusion Ultrasonic method at room temperature can replace conventional refluxing method for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix, and ultrasonic method at heating conditions also can replace conventional refluxing method for content determinations of total flavonoids puerarin from Puerariae Thomsonii Radix. Puerarin contents from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province are all in line with the Pharmacopoeia standards.

Effects of Different Sample Preparation Methods on Total Flavonoids and Puerarin Content from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province / 中国中医药信息杂志

Objective To establish UV spectrophotometry and HPLC methods for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix and Puerariae Thomsonii Radix; To compare the ultrasonic method at room temperature, conventional refluxing method and ultrasonic method at heating conditions at the aspect of content determinations. Methods The content determinations of total flavonoids was determined by UV spectrophotometry at 250 nm; the content of puerarin was determined by HPLC with octadecylsilane-bonded silica gel as the stationary phase, a mixture of methanol and water (25:75) as the mobile phase, 256 nm as the detection wavelength, 1.0 mL/min as the flow rate. Results Contents of total flavonoids in Puerariae Lobatae Radix by ultrasonic method at room temperature, conventional refluxing method and ultrasonic method were15.09%, 14.48%, and 12.71% (n=3), respectively. The contents of puerarin were 4.37%, 4.09%, and 3.80% (n=3), respectively. Contents of total flavonoids in Puerariae Thomsonii Radix were 2.09%, 2.23%, and 2.17% (n=3), respectively. The contents of puerarin were 0.50%, 0.53%, and 0.52% (n=3), respectively. Conclusion Ultrasonic method at room temperature can replace conventional refluxing method for content determinations of total flavonoids and puerarin from Puerariae Lobatae Radix, and ultrasonic method at heating conditions also can replace conventional refluxing method for content determinations of total flavonoids puerarin from Puerariae Thomsonii Radix. Puerarin contents from Puerariae Lobatae Radix and Puerariae Thomsonii Radix in South Anhui Province are all in line with the Pharmacopoeia standards.

Localization of Anterosuperior Point of Transverse-sigmoid Sinus Junction Using a Reference Coordinate System on Lateral Skull Surface / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>During craniotomies using the transpetrosal-presigmoid approach, exposure of the sigmoid sinus remains an essential but hazardous step. In such procedures, accurate localization of the anterosuperior point of the transverse-sigmoid sinus junction (ASTS) is very important for reducing surgical morbidity. This study aimed to create an accurate and practical method for identifying the ASTS.</p><p><b>METHODS</b>On the lateral surfaces of 40 adult skulls (19 male skulls and 21 female skulls), a rectangular coordinate system was defined to measure the x and y coordinates of two points: the ASTS and the squamosal-parietomastoid suture junction (SP). With the coordinate system, the distribution characteristics of the ASTS were statistically analyzed and the differences between the ASTS and SP were investigated.</p><p><b>RESULTS</b>For ASTS-x, significant differences were found in different sides (P = 0.020); the ASTS-x in male skulls was significantly higher on the right side (P = 0.017); there was no significant difference between the sides in female skulls. There were no significant differences in gender or interaction of gender and side for ASTS-x, and for ASTS-y, there were no significant differences in side, gender, or interaction of gender and side. For both sides combined, the mean ASTS-x was significantly higher than the mean SP-x (P = 0.003) and the mean ASTS-y was significantly higher than the mean SP-y (P = 0.011).</p><p><b>CONCLUSIONS</b>This reference coordinate system may be an accurate and practical method for identifying the ASTS during presigmoid craniotomy. The SP might be difficult to find during presigmoid craniotomy and, therefore, it is not always a reliable landmark for defining the ASTS.</p>

Agglutinin isolated from Arisema heterophyllum Blume induces apoptosis and autophagy in A549 cells through inhibiting PI3K/Akt pathway and inducing ER stress / 中国天然药物

Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis (RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume (AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α (p-eIF2α), C/EBP-homologous protein (CHOP), inositol-requiring enzyme 1α (IRE1α), and phosphorylated c-Jun NH-terminal kinase (p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles (AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.