Advances in Modeling of Multiple Myeloma in Mice / 中国医学科学院学报

Multiple myeloma(MM)is a systemic malignancy of plasma cells.Nowadays,the basic research on MM is flourishing with the continuous optimization and innovation of mouse models of MM.Heterologous mouse models of MM established with human-derived cells and immunodeficient mice have been applied in assessing drug efficacy,exploring drug resistance mechanisms,and observing tumor-bone marrow microenvironment interactions.In the last decades,the homologous mouse models of MM established with murine-derived cells or gene-editing technologies have been widely used in the research on the pathogenesis and drug development.Additionally,the stable modeling of targeted organ injury will be a key problem to be tackled in this field.This review summarizes the characteristics and application progress of mouse models of MM.

Efficacy and safefy of Polymyxin B treatment for neutropenic patients suffering from refractory Gram-negative bacterial bloodstream infection / 中华血液学杂志

Objective: To assess the efficacy and safety of polymyxin B in neutropenic patients with hematologic disorders who had refractory gram-negative bacterial bloodstream infection. Methods: From August 2021 to July 2022, we retrospectively analyzed neutropenic patients with refractory gram-negative bacterial bloodstream infection who were treated with polymyxin B in the Department of Hematology of the First Affiliated Hospital of the Soochow University between August 2021 to July 2022. The cumulative response rate was then computed. Results: The study included 27 neutropenic patients with refractory gram-negative bacterial bloodstream infections. Polymyxin B therapy was effective in 22 of 27 patients. The median time between the onset of fever and the delivery of polymyxin B was 3 days [interquartile range (IQR) : 2-5]. The median duration of polymyxin B treatment was 7 days (IQR: 5-11). Polymyxin B therapy had a median antipyretic time of 37 h (IQR: 32-70). The incidence of acute renal dysfunction was 14.8% (four out of 27 cases), all classified as "injury" according to RIFLE criteria. The incidence of hyperpigmentation was 59.3%. Conclusion: Polymyxin B is a viable treatment option for granulocytopenia patients with refractory gram-negative bacterial bloodstream infections.

A combined regimen based on bortezomib and glucocorticoids for 6 patients with recurrent/refractory immune thrombotic thrombocytopenic purpura / 中华血液学杂志

Objective: To observe the efficacy and adverse reactions of a combination therapy regimen based on bortezomib and glucocorticoids in recurrent/refractory immune thrombocytopenic purpura (iTTP) . Methods: Six patients with recurrent/refractory TTP were included and treated with a glucocorticoid and two courses of bortezomib-based regimen. The clinical remission status of patients, changes in ADAMTS13 activity/ADAMTS13 inhibitor, and the occurrence of treatment-related adverse reactions were observed. Results: Of the 6 patients, 2 were males and 4 were females, with a median age of 21.5 (18-68) years. Refractory TTP was found in 1 case and recurrent TTP in 5 cases. Glucocorticoids were administered with reference to prednisone at 1 mg·kg(-1)·d(-1), and gradually reduced in dosage after achieving clinical remission. Bortezomib is subcutaneously administered at 1.3 mg/m(2) on days 1, 4, 8, and 11 with a 28-day treatment course consisting of 2 courses. Six patients achieved clinical remission after receiving bortezomib as the main treatment. ADMATS13 activity returned to normal in all patients with TTP after treatment, and the ADAMTS13 inhibitor turned negative. Thrombocytopenia is the most common adverse reaction after treatment, with other adverse reactions, including peripheral neuritis and abdominal pain, but ultimately all patients returned to normal. In a median follow-up of 26 (9-41) months, 5 patients maintained sustained remission, and 1 patient relapsed after 16 months of bortezomib treatment. Conclusion: Combination therapy of bortezomib and glucocorticoids has a satisfactory therapeutic effect and controllable adverse reactions for recurrent/refractory iTTP.

Absolute bioavailability,dose proportionality,and tissue distribution of rotundic acid in rats based on validated LC-QqQ-MS/MS method / 药物分析学报

Rotundic acid(RA),an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb.(Aquifoliaceae),possesses diverse bioactivities.To further study its pharmacokinetics,a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry(LC-QqQ-MS/MS)method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard(IS).Plasma and tissue samples were subjected to one-step protein precipitation.Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 col-umn(4.6 mm×50 mm,5 μm)under gradient conditions with eluents of methanol:acetonitrile(1∶1,V/V)and 5mM ammonium formate:methanol(9∶1,V/V)at 0.5mL/min.Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode.The developed LC-QqQ-MS/MS method exhibited good linearity(2-500 ng/mL)and was fully validated in accordance with U.S.Food and Drug Administration bioanalytical guidelines.Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral(10,20,and 40 mg/kg)and intravenous(10 mg/kg)administration of RA.Tissue distribution was studied following oral administration at 20 mg/kg.The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1％to 19.4％.RA showed good dose proportionality over a dose range of 10-40 mg/kg.RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver.In conclusion,this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats,which can provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

Study on the mechanism of miR-6751-3p affecting the proliferation and migration of gastric cancer cells / 国际外科学杂志

Objective:To explore the influence of microRNA (miRNA)-6751-3p expression on the proliferation and migration of gastric cancer cells and its molecular mechanism.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-6751-3p in gastric cancer cell lines (MGC803, BGC823, SGC7901, HS-746T) and normal gastric mucosal epithelial cells (GES-1). The gastric cancer cell lines with the lowest expression level of miR-6751-3p were divided into control group and experimental group, and were transfected with miR-NC and miR-6751-3p mimics respectively. qRT-PCR detected the expression level of miR-6751-3p in the two groups of cells. CCK-8 method and scratch healing test were used to detect the proliferation and migration of miR-6751-3p overexpressing cells. The potential target genes of miR-6751-3p were predicted through Deepbase v2.0 and microRNA.org online websites, and the dual luciferase reporter gene experiment was used to verify. qRT-PCR and Western blot were used to detect the expression of target genes in miR-6751-3p overexpression cells.Results:Compared with normal gastric mucosal epithelial cells, the expression of miR-6751-3p was significantly down-regulated in gastric cancer cell lines ( P<0.05), and the cell line with the lowest expression level was MGC803 cells ( P<0.01). Compared with the control group, overexpression of miR-6751-3p can inhibit the proliferation ability ( P<0.05). The scratch healing rate of MGC803 cells in the control group and the experimental group were (65.14±5.65)% and (23.40±6.78)%, respectively. Compared with the control group, the scratch healing rate of MGC803 cells in the experimental group was significantly lower ( t=4.73, P<0.01). The online website predicts that the target gene of miR-6751-3p may be fatty acid binding protein 5 ( FABP5), and miR-6751-3p can complementally bind FABP5 messenger RNA (mRNA) ( t=4.01, P<0.01). Compared with the control group, overexpression of miR-6751-3p can inhibit the expression of FABP5 gene in MGC803 cells ( P<0.01). Conclusion:The expression of miR-6751-3p in gastric cancer cell lines is low, and the overexpression of miR-6751-3p can inhibit the proliferation and migration of gastric cancer MGC803 cells by down-regulating the FABP5 gene.

miRNA-5089-5p inhibits proliferation and migration of esophageal cancer cells by regulating the expression of cathepsin B in vitro / 肿瘤研究与临床

Objective:To investigate the effect of miRNA-5089-5p (miR-5089-5p) on proliferation and migration ability of esophageal cancer in vitro and its relationship with the expression of cathepsin B (CTSB) gene.Methods:Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-5089-5p in 31 tissue samples from patients who underwent esophageal cancer resection and the corresponding pericarcinomatous tissues between in March 2017 and in December 2019 at Huangshi Central Hospital of Edong Healthcare Group, and TE-13, EC9706, Eca109, KYSE30 cell lines and normal esophageal mucosal epithelial HET-1A cells. The esophageal cancer cells with the lowest expression level of miR-5089-5p were divided into 2 groups: miR-5089-5p group transfected with miR-5089-5p mimics and the negative control group with negative control sequence. qRT-PCR was used to detect the expression level of miR-5089-5p after transfection for 48 h. CCK-8 method and scratch healing test were used to detect the proliferation and migration ability of cells in the two groups. The online tools microRNA.org and Deepbase v2.0 were used to predict the target genes of miR-5089-5p. The dual luciferase reporter gene assay was used to verify the target gene of miR-5089-5p. qRT-PCR and Western blot were used to detect the expression level of target genes in the two groups. The expressions of cell proliferation-related protein (PCNA and Ki-67) and migration-related protein (N-Cadherin and Twist) were detected by using Western blot.Results:The relative expression level of miR-5089-5p in esophageal cancer and pericarcinomatous tissues was 1.54±0.53 and 7.07±1.25, respectively ( t = 24.06, P < 0.01). The relative expression level of miR-5089-5p in the esophageal cancer cell lines was lower than that of normal esophageal mucosal epithelial HET-1A cells (all P < 0.05), and the cell line with the lowest relative expression was Eca109 cells (0.12±0.03). Compared with the negative control group, the proliferation ability of Eca109 cells in miR-5089-5p group was gradually reduced with the transfection time extension, and the difference was statistically significant between the two groups since 48 h (all P < 0.05), and the migration ability was also reduced [scratch healing rate: (29±5)% vs.(64±8)%, t=3.91, P < 0.01]. The online tool predicted that the target gene of miR-5089-5p might be CTSB, and the dual luciferase reporter gene assay confirmed that miR-5089-5p complemented CTSB 3'UTR. qRT-PCR results showed that compared with the negative control group, the relative expression level of CTSB mRNA in Eca109 cells of miR-5089-5p group was reduced (0.23±0.04 vs.1.01±0.09, t = 8.27, P < 0.01). Western blot results showed that the expression level of CTSB protein was reduced, and the expression levels of cell proliferation-related protein PCNA, Ki-67 and cell migration-related protein N-Cadherin, Twist were also reduced. Conclusions:The expression level of miR-5089-5p in esophageal cancer tissues and cell lines is low. miR-5089-5p can inhibit proliferation and migration of esophageal cancer Eca109 cells. The mechanism may be achieved by down-regulating CTSB gene expression.

Prevention and control of novel coronavirus pneumonia in non-government primary medical institutions / 中华全科医师杂志

Objective:To understand the coronavirus disease 2019(COVID-19)prevention and control work and the problems and difficulties faced by non-government primary medical institutions in China during the epidemic period.Methods:A survey on the coronavirus disease 2019(COVID-19)prevention and control work of non-government primary medical institutions was conducted on April 14 to 21, 2020 with the self-designed questionnaire. The questionnaire contained three parts: the first part was basic information of medical staff in non-government primary medical institutions, including position and institutional information; the second part was the status quo of non-government primary medical institutions participating in the prevention and control of COVID-19, including the specific work and difficulties faced by the responders during the epidemic period; and the third part was the prevention and control effect of COVID-19 in the responders′ institutions, including whether there were confirmed cases and infected medical staff. An online invitation was issued among the members of General Practice Branch of Chinese Non-government Medical Institution Association. The invited participants included the heads, general practitioners and other medical personnel of the non-government primary medical institutions the invited participants voluntarily scanned the online two-dimensional code to fill in.Results:A total of 761 individuals in primary health institutions from 20 provinces, municipalities and autonomous regions in China participated in the survey. There were 290 (38.1%) men and 471 (61.9%) women with age of 40(32, 48) years; 83.0% (632/761) had worked for more than 5 years; 33.8% (257/761) owned primary professional titles and 33.0% (251/761) owned intermediate titles. Among all participants 28.5% (217/761) were general practitioners, 26.9% (205/761) were institutions/department managers, 14.6% (111/761) were specialists and 40.3% (307/761) were other related personnel. A total of 549 institutions continued to operate during the epidemic period and 96.5% (530/549) participated in the work related to the prevention and control of the epidemic, including prescreening and triage, health consultation, follow-up of suspected patients, donation, quarantine of suspected cases, follow-up of close contacts/discharged patients, diagnosis and treatment of patients with new coronavirus pneumonia. 44.7% (340/761) of respondents participated in the epidemic prevention as front-line staff and directly contacted patients/suspected patients, and 63.1% (480/761) participated in the epidemic prevention work of primary medical institutions, including clinical outpatient service, prescreening triage and screening. The working sites were not limited to the institutions, but also other sites including high-speed railway station. The 97.8% (744/761) responders expressed their willingness to participate in epidemic prevention work under the unified leadership and command of the state. The 63.9% (486/761) of the responders were worried about the lack of protective equipments and measures, and 90.4% (688/761) respondents showed that they needed medical supplies (protective equipment: masks, goggles, protective gowns, etc.).Conclusion:The participation of non-government primary medical institutions and their staff in COVID-19 infection prevention and control is a key component of the epidemic prevention process.

Advantages and disadvantages of common methods of hip-conserving surgery for osteonecrosis of femoral head / 中国组织工程研究

BACKGROUND: There are many methods for the treatment of non-traumatic necrosis of the femoral head. In the clinical treatment, various stages of necrosis are the decisive factors for the choice of operation. In the past, there are many studies about the effectiveness of various techniques, but few studies concern the causes of failure or risk assessment. OBJECTIVE: To explore whether other internal and external factors affect the success rate of hip preservation besides necrosis stage. METHODS: The first author retrieved PubMed, Web of Science, Medline, CNKI and Wanfang database for related studies published from 2010 to 2019. The key words were “osteonecrosis of femoral head, core decompression, non-vascularized bone grafting, vascularized free bone grafting, porous tantalum rod implantation, osteotomy, biological agents, cytotherapy” in English, and “hip-conserving surgery, hip preservation, core decompression, bone graft without blood vessel, bone graft with blood vessel, porous tantalum rod placement, osteotomy, biologics, cell therapy” in Chinese. A total of 214 articles were retrieved, and the full text was consulted. Based on the inclusion and exclusion criteria, 70 articles were finally included for analysis and summary. RESULTS AND CONCLUSION: (1) The advantages and disadvantages of each hip-conserving surgery should be evaluated. (2) Although core decompression is simple, it should not be used alone; the lateral column should be reserved properly when the non-vascularized bone transplantation is used to clean up the dead bone, so as to deal with the mechanical weakening; the blood loss and fracture risk are higher when the vascularized bone transplantation is converted to total hip replacement; the stress concentration is easily caused by tantalum rod implantation, and the clinical application is less. (3) Osteotomy, as a kind of hip-conserving surgery with great trauma, should be planned in detail for patients’ age, body mass index, and necrotic range, besides considering the necrotic stage. (4) The risk of total hip replacement should be considered in the long run no matter which operation. (5) The combination of various surgical methods and biological agents may achieve better results.

Research Progress of Polysaccharide Utilization Loci and Its Application in Pharmacological Research of Chinese Medicinal Herb / 中国实验方剂学杂志

Polysaccharide utilization loci (PULs) are a group of gene clusters related to polysaccharides catabolism and located in specific areas, including starch utilization system(SusC), SusD, encoding outer membrane glycoprotein binding proteins, and carbohydrate active enzymes. The wide existence of PULs in Bacteroides, and the extraordinary ability of utilizing polysaccharide is its survival strategy to adapt to the intestinal tract. On one hand, Bacteroides feature a high abundance and variety, making it the most important bacterial target group regulated by traditional Chinese medicine(TCM). On the other hand, polysaccharide is an important effective constituent among TCM chemicals, with a high content, and can also be used as a competitive carbon source for intestinal bacteria. Therefore, what is the mechanism of regulating the intestinal flora based on the carbon source of polysaccharide is an important part of the pharmacological research of TCM. According to the latest literatures, this paper introduces the gene and protein composition of PULs, reviews the latest developments in PULs research, and analyzes the structure of PULs in the genomes of Bacteroides fragilis and Bacteroides thetaiotaomicron. Furthermore, we also put forward a prospect for the pharmacological micro-ecological mechanism of TCM based on PULs based on our carbon source experiments, which focuses the effects of Bacteroides by TCM polysaccharides in vitro. This research is not only the new content of bacterial PULs researches, but also the important part of researches of "human-drug-bacteria" holistic view and TCM spleen-tonifying concept.

Effect of Yiqi Huoxue Recipe on Oxidative Stress Response in Rats with Cerebral Ischemia Injury Based on Nrf2/HO-1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the protective effect of Yiqi Huoxue recipe on rats with cerebral ischemia injury by using oxidative stress injury as an entry point. Method:SD rats were randomly divided into model group, sham operation group, nimodipine group (20 mg·kg-1), Yiqi Huoxue recipe high, medium and low dose group (2.916,1.458,0.729 g·kg-1). After 14 days of stomach, acute cerebral ischemic injury model was established by ligation of bilateral common carotid arteries. Ultrasound of synapse was observed by transmission electron microscopy. Total superoxide dismutase (T-SOD) and dialdehyde (MDA), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), horizontal adenine nucleoside triphosphate (ATP) levels were detected by biochemical method. Western blot and Real-time PCR was used to determine the expression of heme oxygenase-1(HO-1) and nuclear factor E2-related factor 2 (Nrf2) protein and mRNA in the ischemic cortex of rats. Result:Transmission electron microscopy showed that Yiqi Huoxue recipe had a significant improvement on the degree of cerebral ischemic injury. Compared with sham operation group, MDA levels in the brain homogenate of model group increased significantly, T-SOD and GSH-Px levels were significantly decreased (P+-K+-ATP ase, Ca2+-Mg2+-ATP ase and ATP was significantly decreased (PPPPP+-K+-ATP ase, Ca2+-Mg2+-ATP ase and total ATP activity(PPPPPPConclusion:Yiqi Huoxue recipe may protect against cerebral ischemic injury by inhibiting oxidative stress through Nrf2/HO-1 signaling pathway.

The determination of diflucortolone in rabbit plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS) / 药学学报

A sensitive and efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for quantitative determination of diflucortolone in rabbit plasma after dermal administration of diflucortolone valerate cream to rabbits. After extraction with ethyl acetate, the chromatographic separation was performed on Zorbax Eclipse XDB-C18 (50 mm×4.6 mm, 5 μm) with a gradient mobile phase consisting of 50% acetonitrile-50% methanol and 0.1% formic acid-5% methanol-5 mmol·L-1 ammonium formate at a flow rate of 0.35 mL·min-1. The quantitative analysis was carried out using multiple reaction monitoring (MRM) at specific ion transitions of m/z [M+H]+ 395.2→m/z 355.2 for diflucortolone and m/z [M+H]+ 258.1→m/z 120.9 for ethoxyphenylethylamine (internal standard) in positive ion mode with electrospray ionization (ESI) source. This validated LC-MS/MS method had a linearity over the concentration range of 0.01-10 ng·mL-1 with the lower limit of quantification (LLOQ) at 0.01 ng·mL-1. At level of LLOQ, the inter and intra-assay precision (RSD) were no greater than 9.82% and 11.0%, respectively. The main pharmacokinetic parameters of the diflucortolone including tmax, Cmax, AUC0-72 h, and t1/2 were as follows: (6.33±1.21) h, (0.168±0.080 0) ng·mL-1, (3.15±0.834) h·ng·mL-1, (32.0±17.4) h. The method was validated in the pharmacokinetic study of diflucortolone in rabbit following dermal administration of diflucortolone valerate cream at dose of 0.01 g·cm-2. In this study, the program of animal testing had been approved by Committee on the management and usage of experimental animal in the Evaluation Company of Innovative Drug, Tianjin Institute of Pharmaceutical Research.

Summary and core technique on the origin and evolution of reinforcing-reducing manipulation achieved by lifting and thrusting the needle / 中国针灸

By analyzing the evolution of reinforcing-reducing manipulation achieved by lifting and thrusting the needle recorded in ancient literature of traditional Chinese medicine, it is found that the main contents of reinforcing-reducing manipulation by lifting and thrusting the needle include manipulating speed change, manual amplitude, insertion layer, gender, the direction to the acupuncture receiver, forenoon and afternoon and relevant quantity. Among them, gender, the direction to the acupuncture receiver, forenoon and afternoon and relevant quantity are the unnecessary parameters, while the manipulating speed change, manual amplitude and insertion layer are the indispensable parameters. The manipulating speed change is the core of the necessary parameters for the reinforcing-reducing manipulation achieved by lifting and thrusting the needle. Combined with the manual amplitude, the manipulating speed of needle determines the volume of needling stimulation. The insertion layer is decided on the base of the clinical demand. In the core technique of reinforcing-reducing manipulation by lifting and thrusting the needle, the reinforcing is achieved by thrusting the needle forcefully and quickly and then lifting the needle body slowly and evenly back to the original layer. The reducing is achieved by lifting the needle forcefully and quickly and then thrusting the needle body slowly and evenly back to the original layer. The manipulating speed and manual amplitude of needling are the parameters to quantize acupuncture manipulation. In association with the acupuncture effects in human body, these parameters contribute to the interpretation of the dose-effect relationship of acupuncture and the improvement of clinical effects.

Analysis of diagnosis and treatment of solid pseudopapillary neoplasm of pancreas in 12 children / 中华实用儿科临床杂志

Objective To analyze the experience in diagnosis and surgical treatment for solid pseudopapillary neoplasm(SPN) of pancreas in children.Methods A retrospective study was performed in 12 pediatric patients with SPN who had been admitted to Affiliated Cancer Hospital of Zhengzhou University during January 2004 to December 2016,and their general data,demographic data,types of operations,postoperative complications and follow-up were analyzed.Results Among the 12 patients,3 cases were male and 8 cases were female,with average age 14.3 years old (11-17 years old).The main clinical manifestations included abdominal pain(4/12 cases,33.3％),abdominal mass (2/12 cases,16.7％) and trauma(2/12 cases,16.7％).In those 12 patients,33.3％ (4/12 cases) SPN was located at the head of the pancreas,and 66.7％ (8/12 cases) at the body and tail of it.The tumors were usually large,the largest diameter ranged from 4.0 to 15.3 cm(average largest diameter,8.2 cm).The color uhrasonography indicated heterogeneous echogenic mass and clear boundary.CT scanning indicated that the tumor was a low-density cystic mass with a clear boundary,with enhanced tumor real component and irregular reinforcement.No calcification was found in the patients.Dynamic enhanced magnetic resonance imaging scan revealed gradual strengthening solid components in tumor.All the patients received surgical resection,with distal pancreatectomy in 4 patients,pancreaticoduodenectomy in 4 patients,spleen-preserving distal pancreatectomy in 2 patients,Enucleation in 1 patient,and distal pancreatectomy and self-splenic slices transplantation in 1 patient.Lymphadenectomy was performed in 4 patients,and all the 21 removed lymph nodes were all negative.Pathological diagnosis confirmed the SPN in all the patients,among them 3 cases were malignant SPN,and one of them with tumor rupture and hemorrhage.The mean follow-up duration was 57.7 months(19-156 months) and no recurrence was found.Conclusion SPN is a rare neoplasm in children who go to see doctors because of clinical symptoms.Surgical resection,especially organs-preserving resection,may improve the long-term results.

Effects of Long-term Sleep-deprivation on Rats′Learning and Memory Behavior and Neurotransmitters in Hippocampus and the Effects of Prevention of Electro-acupuncture on the Sishencong Acupoints / 中山大学学报(医学科学版)

[Objective]To investigate the effects of continuous long-term sleep deprivation on rat's behavior,memory and neurotransmitters in hippocampus,as well as determine the effects of prevention of electro-acupuncture.[Method]We used multi-platform water environment to make the rat model of insomnia.The experiment was divided into two sections.Sec-tion one:thirty rats were divided into 5 groups(six rats each group):tank control group(TC),sleep-deprived for continu-ous 7 days group(SD group),three days of recovery after SD group(3d after SD group),six days of recovery after SD group (6d after SD group),eleven days of recovery after SD group(11d after SD group).The term of recovery refers to normal feed-ing and sleeping when rats return to cages after sleep-deprivation.The open-field test was performed following the end time of each group experiment and then the rats were perfused and fixed.Section two:Eighteen rats were divided into three groups (six rats each group):tank control group(TC),sleep-deprived for continuous 7 days group(SD group),sleep-deprived rats treated with electro-acupuncture group(SD+EA group).Rats of SD group and SD+EA group were deprived sleep for con-tinuous 7 days. During the period of sleep deprivation,the rats of SD+EA group were treated with electro-acupuncture 20 minutes every morning.At the end time of experiment,rats were tested by the Morris water maze and then perfused and get specimens.[Results]Section one:the statistical analysis of outcome of open-field test showed:as compared to the TC group,the scores of rat's horizontal movement and vertical movement were significantly fewer in the SD group(P<0.05).Af-ter 3,6,or 11 days recovery of sleep,the scores increased significantly as compared with SD group(P<0.05). Although, the outcome of open-field test in the 3d after SD and 6d after SD groups had the significant difference with TC group(P<0.05),there was no different between the 11d after SD group and TC group(P>0.05).The quantitative analysis of gamma aminobutyric acid(GABA)expression in the hippocampus showed:the SD group shows lower level of GABA when com-pared to the TC group(P<0.05).After 3 days or 6 days of recovery,the expression of GABA is higher than the SD group(P<0.05),and similar to the TC group(P>0.05).In the 11days after SD group,the expression of GABA declined as compared to the TC group and the 6d after SD group(P<0.05).Section two:In the Morris water maze experiment,when compared to the TC group and the SD+EA group,the escape latency period was significantly prolonged and the times of across the plat-form were significantly reduced in the SD group(P<0.05).On the contrary,there were no statistical differences in the escape latency period or the across platform times between the SD+EA group and the TC group(P>0.05).The results of serotonin(5-HT)and GABA in the hippocampus showed:the 5-HT level in the SD group was no significant difference as compared to the TC group and the SD group on 5-HT level,while the 5-HT level in the SD+EA group was reduced as compared to the TC group(P<0.05). In the opposite,the GABA level in the SD group was decreased as compared to the TC group(P<0.05),while the SD+EA group and the TC group appear similarly level(P>0.05).[Conclusions]The effect of 7 days'contin-uous long-term sleep-deprivation may lead to the reduction in the ability of learning and memory behavior of rats.Our results is different with 4 days'continuously sleep-deprivation which was reported on the rats'expression of GABA and 5-HT. Electro-acupuncture stimulating the Sishengcong Acupoints seems preventing the harming of long-term sleep-deprivation on the ability of learning and memory,which probably is through adjusting the disorder of central neurotransmitter expression caused by sleep deprivation.

Effects of FEA from Polygonum hydropiper L. on Biochemical Indexes and Inflammatory Cytokines in Mice with Endotoxemia Induced by Lipopolysaccharide / 中国中医药信息杂志

Objective To observe the effect of flavonoids ethyl acetate(FEA)from Polygonum hydropiper L.on biochemical indexes and inflammatory cytokines in mice with endotoxemia; To expore the mechanism. Methods Total flavonoids in the whole plant of Polygonum hydropiperum L. were extracted by enzymolysis-ultrasonic coupling method. The FEA part were obtained by extracting and separating, followed with macroporous resin purification and enrichment. The animal model of endotoxemia was established by stimulation of lipopolysaccharide (LPS) in mice. Experimental mice were divided into blank group, model group, FEA high-, medium-, and low-dose groups. Each administration group was given the corresponding concentration of herb liquor. The concentration of malondialdehyde (MDA) and myeloperoxidase (MPO) in intestinal tissue, total antioxidant capacity (T-AOC), superoxide dismutase (T-SOD) activity and glutathione peroxidase (GSH-Px) activity in liver tissue, glutathione (GSH), lysozyme (LZM) and acid phosphatase (ACP) in serum were determined. The contents of tumor necrosis factor-α (TNF-α), interferon (IFN)-α, IFN-γ and interleukin-2 (IL-2) in lung tissue were detected by RT-PCR. Results Compared with blank group,the levels of MDA, MPO in intestinal tissue and serum ACP of model mice were increased, while T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM levels were decreased; TNF-α in serum, intestinal and liver tissues were increased, the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissue were increased. Compared with the model group, the levels of MDA, MPO in intestinal tissue and serum ACP were decreased in all dose of FEA groups;The levels of T-AOC, T-SOD, GSH-Px in liver tissue, serum GSH and LZM of FEA medium and high-dose groups were increased. The content of TNF-α in mice serum, intestinal and liver tissues of all dose of FEA groups were significantly reduced, and the expressions of TNF-α, IFN-α, IFN-γ and IL-2 mRNA in lung tissues were significantly decreased. The pathological morphology of mice lung, ileum and colon tissues of FEA high-dose group were significantly ameliorated than model group. Conclusion FEA can attenuate inflammation injury of endotoxemia mice induced by LPS, which has protective effects for organism.

Metabonomics delineates allergic reactions induced by Shuang-huang-lian injection in rats using ultra performance liquid chromatography-mass spectrometry / 中国天然药物

Shuang-huang-lian Injection (SHLI) is the first successfully developed drug from traditional Chinese medicine (TCM) powder for injection, since its use for the treatment of acute respiratory tract infection, pneumonia, influenza, etc. At the same time, its allergic reactions have also emerged, which limits clinical applications. However, few scholars pay attention to the mechanism of allergic reactions. In this present study, metabonomics technology was used to explore the changes in endogenous metabolites in urine of the rat model of SHLI induced allergic reaction; we and analyzed the metabolites, metabolic pathway, and the mechanism which were closely related to the allergic reactions. The levels of serum histamine and tryptase were examined and changes in histomorphology were also observed. Based on the UPLC-Q-TOF/MS metabonomics, we carried out the pattern recognition analysis, selected potential biomarkers associated with allergic reactions, and explored the pathological mechanism for SHLI induced allergic reaction, which laid the foundation for the safety research of SHLI. Our results showed that SHLI increased the levels of serum histamine and tryptase in rats with allergic reaction; we determined 15 biomarkers in rat allergic reaction model induced by SHLI and found multiple metabolic pathways involved, such as metabolism of linolenic acid, phenylalanine, amino acid, 2-oxo acid, and purine and other metabolic pathways.

Determination of obeticholic acid in rat plasma by liquid chromatography-tandem mass spectrometry (LC-MS/MS) / 药学学报

A simple and sensitive method was developed for quantitation of obeticholic acid in rat plasma with liquid chromatography-tandem mass spectrometry (LC-MS/MS). After liquid-liquid extraction by methyl tert-butyl ether, the chromatographic separation was carried out on an ACE Excel 2 Super C18 column (50 mm×2.1 mm ID, 1.7 μm) with a gradient mobile phase consisting of acetonitrile and 2 mmol·L-1 ammonium formate at a flow rate of 0.2 mL·min-1. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 418.9[M-H]-→401.2 for obeticholic acid and m/z 469.0[M-H]-→ 425.2 for glycyrrhetinic acid (internal standard) in the negative ion mode with electrospray ionization (ESI) source. This validated LC-MS/MS method yielded a good linearity over the range of 5 -5 000 ng·mL-1 with the lower limit of quantitation (LLOQ) of 5 ng·mL-1. The intra and inter-assay precisions (RSD) were all less than 9.82% and the accuracy (RE) was within ±6.90%. The extraction recovery of obeticholic acid was from 85.4% to 88.5%, and the matrix effect of obeticholic acid ranged from 78.9% to 82.5%. Stability test suggest that obeticholic acid in rat plasma was stable for 24 h on workbench, up to 1 month at -70℃, and after three cycles of freeze-thaw. Extracted samples were stable for more than 24 h in an auto-sampler at 6℃. The precision was less than 7.25%, and the accuracy was within ±11.2%, after being diluted 10 times by blank rat plasma. The method has been successfully applied to a pharmacokinetic study of obeticholic acid in rats following oral administration at the dose of 2.5 mg·kg-1.

Metabonomics delineates allergic reactions induced by Shuang-huang-lian injection in rats using ultra performance liquid chromatography-mass spectrometry / 中国天然药物

Shuang-huang-lian Injection (SHLI) is the first successfully developed drug from traditional Chinese medicine (TCM) powder for injection, since its use for the treatment of acute respiratory tract infection, pneumonia, influenza, etc. At the same time, its allergic reactions have also emerged, which limits clinical applications. However, few scholars pay attention to the mechanism of allergic reactions. In this present study, metabonomics technology was used to explore the changes in endogenous metabolites in urine of the rat model of SHLI induced allergic reaction; we and analyzed the metabolites, metabolic pathway, and the mechanism which were closely related to the allergic reactions. The levels of serum histamine and tryptase were examined and changes in histomorphology were also observed. Based on the UPLC-Q-TOF/MS metabonomics, we carried out the pattern recognition analysis, selected potential biomarkers associated with allergic reactions, and explored the pathological mechanism for SHLI induced allergic reaction, which laid the foundation for the safety research of SHLI. Our results showed that SHLI increased the levels of serum histamine and tryptase in rats with allergic reaction; we determined 15 biomarkers in rat allergic reaction model induced by SHLI and found multiple metabolic pathways involved, such as metabolism of linolenic acid, phenylalanine, amino acid, 2-oxo acid, and purine and other metabolic pathways.

Survival analysis of patients with neutropenia and advanced gastric cancer treated with chemotherapy / 中国综合临床

Objective To investigate the effect of neutropenia on the quality of life in patients with advanced gastric cancer after chemotherapy.Methods All patients were treated with FOLFOX6 as initial chemotherapy after the diagnosis,14 days as a cycle,the treatment time had 4 cycles.After 4 cycles of chemotherapy,neutrophil levels were measured in all patients.According to CTCAE 4.0 standard.All patients were divided into the non neutropenia reduction group(the non reduction group,17 cases),the mild neutropenia reduction group(the mild reduction group,44 cases)and the severe neutropenia reduction group(the severe reduction group,35 cases).According to RECIST1.1 criteria,the clinical efficacy of the three groups were evaluated.The objective response rate(ORR)and disease control rate(DCR)of the three groups were calculated,and the progression free survival(PFS)and overall survival(OS)were compared among the three groups.Results The difference in clinical efficacy among the non reduction group,the mild reduction group and the severe reduction group was statistically significant(U=15.2314,P=0.0000).The objective response rates in the non reduction group,the mild reduction group and the severe reduction group were 17.65%,56.82%,and 25.71%(χ2=11.7694,P=0.0028),and the disease control rates of the patients in the three group were 47.06%,84.09% and 54.29%,respectively(χ2 = 11.3668,P= 0.0034).The objective response rate and disease control rate in the mild reduction group were significantly higher than those in the non reduction group and the severe reduction group,and the differences among the three groups were statistically significant(P<0.05).At the same time,there were significant differences among the three groups in PFS and OS((F=6.56, 11.11,P<0.05).The PFS and OS in the mild reduction group were significantly higher than those in the non reduction group and the severe reduction group(P<0.05).Conclusion Patients with advanced gastric cancer may have better short-term clinical efficacy and survival time due to mild neutropenia caused by chemotherapy.

Determination of halometasone and its pharmacokinetics study in rabbit plasma by LC-MS/MS / 中国药理学通报

Aim TodevelopandvalidateaLC-MS/MS assay to quantify halometasone in rabbit plasma and study pharmacokinetics of halometasone after dermal topical administration of Halometasone Cream.Meth-ods Theplasmasamplewassubmittedtoliquid-liquid extraction using methyl tertiary butyl ether,with dexa-methasone as the internal standard (IS ).Chromato-graphic separations were performed on a Diamonsil C18 column(100 mm ×4. 6 mm,5 μm)with a linear gra-dient of methanol and 2 mmol · L-1 ammonium ace-tate.Halometasone and dexamethasone(IS)were ion-ized with an ESI source operated in negative ion mode, and the detected ions were m/z 503. 1→413. 0 (halo-metasone),m/z 391. 0→361. 0 (dexamethasone ). The test article could be monitored in rabbit plasma when following single dermal topical administration of Halometasone Cream at 1 g/100 cm2 to rabbits by u-singavalidatedLC-MS/MSassay.Results Calibra-tion curve was linear over the concentration range of0. 02~20 μg·L-1 in rabbit plasma.For low,medi-um,high concentration of QC solutions,the intra-and inter-day precision was in the range of 3. 72% ~7. 87%, and the accuracy was within 99. 1% to 103%. The pharmacokinetic parameters in rabbits were as follows:Tmax,Cmax,AUC0-t,T1/2 was (7. 38 ± 1. 06)h,(1. 16 ±0. 527)μg·L-1,(18. 8 ±7. 23)h·μg·L-1 ,(13. 8 ±3. 70)h,respectively.Conclusions ThisLC-MS/MSanalysismethodhashighsensitivi-ty,and sample processing method is simple,which has been rigorously validated.The method could be suc-cessfully applied to the pharmacokinetic study of halo-metasone after skin administration of Halometasone Cream to rabbits.