Effect of Morindae Officinalis Radix Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Improvement of Renal Function and HPG Axis of Kidney Yang Deficiency Model Rats Induced by Adenine / 中国实验方剂学杂志

ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.

Effect of Morindae Officinalis Radix Processed with Different Proportions of Glycyrrhizae Radix et Rhizoma on Improvement of Renal Function and HPG Axis of Kidney Yang Deficiency Model Rats Induced by Adenine / 中国实验方剂学杂志

ObjectiveTo explain the scientific connotation of Morindae Officinalis Radix （MOR） processed by Glycyrrhizae Radix et Rhizoma （Gly） by comparing the effect of raw products of MOR and processed products of MOR with different proportions of Gly （GMOs） on the improvement of renal function and hypothalamic-pituitary-gonadal （HPG） axis， the protein expression of Wnt/β-catenin and transforming growth factor-β1 （TGF-β1）/Smad signal pathways in kidney Yang deficiency model rats induced by adenine. MethodGMOs were prepared according to method under MOR in 2020 edition of Chinese Pharmacopoeia. Rat model of kidney Yang deficiency was established by intragastrical administration of adenine， levels of follicle stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2） and testosterone （T） were measured by enzyme-linked immunosorbent assay （ELISA）. Levels of urea nitrogen （BUN） and serum creatinine （SCr） were measured by spectrophotometry， hematoxylin-eosin （HE） staining was used to evaluate the pathological changes of kidney， testis and epididymis. Immunohistochemistry （IHC） was used to analyze the protein expression of E-cadherin， α-smooth muscle actin （α-SMA）， Wnt2b， β-catenin， Smad1 and Smad4. ResultMOR processed with 100∶6 and 100∶12 proportions of Gly （short for GMO/100∶6 and GMO/100∶12） had the most obvious improvement on the body posture of kidney Yang deficiency model rats. GMO/100∶12 had the best effect on reducing the levels of BUN， SCr， FSH， LH and the ratio of E2/T. GMO/100∶6 and GMO/100∶12 had the best effect on regulating the protein expression of E-cadherin， α-SMA， Wnt2b， β-catenin， Smad1 and Smad4. ConclusionGMO/100∶6 and GMO/100∶12 have the a good effect on the improvement of renal function and HPG axis in kidney Yang deficiency model rats induced by adenine， which is related with the fact that they can regulate Wnt/β-catenin pathway in renal and testicular tissue and TGF-β1/Smads pathway in testicular tissue.

Flexion-lateral curvature-supination reduction for treatment of lower cervical dislocation with unilateral facet interlocking / 中华创伤骨科杂志

Objective:To investigate the efficacy of flexion-lateral curvature-supination reduction combined with primary anterior surgery for the treatment of lower cervical dislocation with unilateral facet inter-locking.Methods:A retrospective analysis was performed in the 32 patients who had been admitted to Department of Spine Surgery, Honghui Hospital for lower cervical dislocation with unilateral facet interlocking from November 2015 to October 2018. According to their treatments, they were divided into 2 groups. In the emergency group treated by flexion-lateral curvature-supination reduction combined with primary anterior surgery, there were 13 males and 3 females, aged from 24 to 63 years. In the traction group treated by cranial traction reduction combined with secondary anterior surgery, there were 12 males and 4 females, aged from 20 to 64 years. The operation time, intraoperative blood loss, hospital stay, bone graft fusion, American Spinal Injury Association (ASIA) grade and Japanese Orthopaedic Association (JOA) score were compared between the 2 groups.Results:There was no significant difference in the preoperative general data between the 2 groups, showing they were comparable ( P<0.05). All patients were followed up from 26 to 40 months. The hospital stay for the emergency group [(7.2±1.2) d] was significantly shorter than that for the traction group[(10.9±1.2) d] ( P<0.05). There was no significant difference in the operation time, blood loss, ASIA grade or JOA score between the 2 groups ( P>0.05). All patients achieved osseous fusion of intervertebral space. Conclusion:Compared with traditional methods, flexion-lateral curvature-supination reduction combined with primay anterior surgery shows no significant difference in the recovery of neurological function but leads to a shorter hospital stay.

Comparison of curative effect of forceful reduction percutaneous pedicle screw and ordinary percutaneous pedicle screw in the treatment of osteoporotic thoracolumbar burst fracture with kyphosis / 中华创伤杂志

Objective:To compare the efficacy of forceful reduction percutaneous pedicle screw and ordinary percutaneous pedicle screw in the treatment of osteoporotic thoracolumbar burst fracture with kyphosis.Methods:A retrospective cohort study was conducted to analyze the clinical data of 566 patients with osteoporotic thoracolumbar burst fracture with kyphosis admitted to Honghui Hospital, Xi ′an Jiaotong University from January 2015 to December 2018, including 191 males and 375 females, with age range of 48-79 years [(61.7±10.7)years]. Fracture segments were located at T 11 in 134 patients, T 12 in 154, L 1 in 160, and L 2 in 118. All fractures were type IIIA according to the acute symptomatic osteoporotic thoracolumbar fracture classification (ASOTLF). The thoracolumbar osteoporotic fracture severity score assessment system (TLOFSAS) score was ≥5 points. A total of 275 patients underwent forceful reduction and percutaneous pedicle screw internal fixation (forceful reduction screw group), and 291 patients underwent common percutaneous pedicle screw internal fixation (common screw group). The operation time, intraoperative blood loss, times of X-ray exposure on patients and measures documented before operation, at 3 days after operation and at 2 years after operation including anterior height ratio of the injured vertebrae, sagittal Cobb angle of the injured vertebrae, Japanese Orthopaedic Association (JOA) score and visual analog scale (VAS) were compared between the two groups. Moreover, degree of correction of Cobb angle at 3 days after operation, loss of correction of Cobb angle at 2 years after operation and postoperative complications were observed. Results:All patients were followed up for 25-34 months [(29.9±3.4)months]. The operation time, intraoperative blood loss and times of X-ray exposure on patients in forceful reduction screw group were (69.4±10.2)minutes, (60.3±13.1)ml and (26.8±3.7)times, less than (80.6±11.9)minutes, (80.7±15.4)ml and (30.4±3.4)times in common screw group (all P<0.01). There was no significant difference in anterior height ratio of the injured vertebrae between the two groups before operation and at 3 days after operation (all P>0.05). The anterior height ratio of the injured vertebrae in forceful reduction screw group was (95.5±2.3)% at 2 years after operation, significantly higher than (85.4±1.7)% in common screw group ( P<0.01). There was no significant difference in sagittal Cobb angle of the injured vertebrae between the two groups before operation ( P>0.05). The sagittal Cobb of the injured vertebrae in forceful reduction screw group at 3 days and 2 years after operation were (7.9±1.6)° and (8.8±1.5)°, lower than (10.6±1.1)° and (12.3±1.2)° in common screw group ( P<0.05 or 0.01). There were no significant difference in JOA score and VAS between the two groups before operation, at 3 days and at 2 years after operation (all P>0.05). The degree of correction of Cobb angle in forceful reduction screw group was (19.4±2.5)°, higher than (17.3±2.6)° in common screw group ( P<0.05). The loss of correction of Cobb angle in forceful reduction group was less than that in common screw group at 2 years after operation, but the difference was not statistically significant ( P>0.05). The incidence of postoperative complications in forceful reduction screw group was 12.4% (34/275), compared to 14.1% (41/291) in common screw group ( P>0.05). There were no complications such as iatrogenic nerve injury, fracture or loosening of internal fixator or leakage of bone cement in the spinal canal in both groups. Conclusions:For osteoporotic thoracolumbar burst fracture with kyphosis, forceful reduction and percutaneous pedicle screw internal fixation can significantly shorten operation time, reduce intraoperative blood loss and times of X-ray exposure on patients, restore height of the injured vertebrae, correct kyphosis and maintain reduction height of the injured vertebrae in contrast with conventional percutaneous pedicle screw internal fixation.

Evaluation of ß-agonists in blood meal: Validation of determination method and potential pathway for reentry into the environment.

ß-agonists, which have been illegally used in animal production in some countries, can induce bioaccumulation when blood is converted by rendering into blood meal. Unfortunately, available data on this topic are scarce, which result in lack of risk assessment. Therefore, in this research, a method for simultaneous determination of 22 ß-agonists in blood meal by liquid chromatography coupled with tandem mass spectrometry using isotope dilution was developed. The recoveries of the developed method ranged from 68.6% to 118.8% with RSD at below 20%. the limit of detection (LOD) is blew 1 µg/kg. The change in agonist form added and incurred blood into blood meal and long stability of ß-agonist in blood meal were studied. Then, we analyzed blood meal for 22 agonists using this method. The results suggest blood meal is a possible pathway for agonist reentry into animals. Potential risks of agonist residues in blood meal were examined. This study is the first to explore source of ß-agonist residues in blood meal, change in processing produce and stability in stored stage.

Simultaneous determination of antibiotics and amantadines in animal-derived feedstuffs by ultraperformance liquid chromatographic-tandem mass spectrometry.

In this study, a method for simultaneous determination of 40 antibiotics from 6 different drug families (7 quinolones, 5 penicillins, 8 macrolides, 2 lincosamides, 4 tetracyclines, and 13 sulfonamides) and 3 amantadines in animal-derived feedstuffs (ADF) through ultraperformance liquid chromatography-tandem mass spectrometry using a new lipid-removing solid-phase extraction column PRiME HLB was developed. The sample was extracted with 20 mL extraction solution and then purified and concentrated with PRiME HLB. The eluent was evaporated to dryness under nitrogen and analyzed with LC-MS/MS using acetonitrile and 0.1% formic acid as the mobile phase via gradient elution. Under the optimum conditions, recoveries were 65.8% to 104.4% with a relative standard deviation of <15% at spiked levels of 5.0 µg/kg to 100 µg/kg. The limits of detection ranged from 0.5 µg/kg to 5 µg/kg. 15 of the 43 drugs were found in ADF. Sulfonamides, fluoroquinolones, macrolides, tetracyclines, and amantadines were detected, with the highest levels reaching 325 µg/kg.

Accumulation and Determination of Phenylethanolamine A Residue in Hair of Swine and Sheep.

The present study proposed the use of liquid chromatography-tandem mass spectrometry to detect the novel ß-agonist phenylethanolamine A (PEA) in incurred hair samples of swine and sheep and to assess its accumulation for residue monitoring. The method showed good percent recoveries ranging from 93.2% to 102% and good coefficient of variation at <15%. The experiment was conducted in swine (24 treated and 4 controls) and sheep (3 treated and 1 control). PEA concentration was determined in hair during the treatment. High residue concentrations were present in hair as early as Day 24 (14.8 ± 3.6 and 25.8 ± 7.6 ng/g) and Day 21 (23.4 ± 6.6 ng/g) for swine and sheep; these residues persisted until withdrawal on Days 14 and 21. Results showed high PEA accumulation in hair, thereby indicating the use of hair as a matrix in the control of PEA abuse in farm animals.

Simultaneous Detection of 21 Kinds of Mycotoxins and Their Metabolites in Animal Plasma with Impurity Adsorption Purification Followed by Liquid Chromatography-Tandem Mass Spectrometry / 分析化学

A novel method for simultaneous detection of mycotoxins (e.g.,aflatoxin B1) or their metabolic residues in animal plasma with impurity adsorption purification followed ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed.Extraction of mycotoxins and their metabolites from animal plasma sample was performed with 0.1％ formic acid-acetonitrile solution after addition of sodium chloride and hydrous magnesium sulfate.The extract was then dehydrated and purified with hydrous magnesium sulfate,C18,primary secondary amine,and alumina-A.3 mL of the supernatant was evaporated and re-dissolved with 0.5 mL of 0.1％ formic acid aqueous solution/acetonitrile (70∶ 30,V/V) for UPLC-MS/MS detection.The analytes were separated by a C18 column utilizing gradient elution with 0.1％ formic acid aqueous solution containing 0.5 mmol of ammonium acetate and 0.1％ formic acid-methanol solution,and finally detected by tandem mass spectrometry in positive/negative ESI mode.Identification and quantification were achieved by LC-MS/MS with multi-reaction monitoring (MRM).Good linearity in response was obtained in the analytes concentration range of 0.05-100 ng/mL with correlation coefficients larger than 0.99.The limits of quantification (S/N=10) were around 0.05-0.5 ng/mL.The recoveries of mycotoxins and their metabolites spiked in blank plasma samples were in the range of 62.0％-116.4％,with relative standard deviations (RSDs) less than 19.0％.

mTOR: An attractive therapeutic target for osteosarcoma?

Osteosarcoma (OS) is a common primary malignant bone tumor with high morbidity and mortality in children and young adults. How to improve poor prognosis of OS due to resistance to chemotherapy remains a challenge. Recently, growing findings show activation of mammalian target of rapamycin (mTOR), is associated with OS cell growth, proliferation, metastasis. Targeting mTOR may be a promising therapeutic approach for treating OS. This review summarizes the roles of mTOR pathway in OS and present research status of mTOR inhibitors in the context of OS. In addition, we have attempted to discuss how to design a better treatment project for OS by combining mTOR inhibitor with other drugs.

Determination of 26 Mycotoxins in Feedstuffs by Multifunctional Cleanup Column and Liquid Chromatography-Tandem Mass Spectrometry / 分析化学

A rapid high-throughput method for the determination of 26 mycotoxins involving multifunctional cleanup column coupled with liquid chromatography-tandem mass spectrometry ( LC-MS/MS) was developed and validated for the determination in feedstuffs. The feedstuff samples were extracted by ultrasonic treatment for 1 hour and the extraction solvent was acetonitrile/water/formic acid (84:15. 9:0. 1, V/V). 1 mL of the supernatant layer was purified by a commercial Mycospin 400 multifunctional cleanup column, then dried and re-dissolved by 0. 25 mL water/methanol/formic acid (95:4. 9:0. 1, V/V) in a vial for injection into the LC-MS/MS system. Chromatographic analyses were carried out on a reversed phase C18 column and using a gradient elution with 0. 1% formic acid aqueous solution and 0. 1% formic acid methanol solution. The mass spectrometer was operated in a multiple reaction monitoring ( MRM) mode that selected one precursor ion and two product ions for each target compound. Validation studies were carried out in maize and soybean meal as representative matrixes. The most target compounds had different level of matrix effects. So, matrix-matched calibration was adopted for quantification. Mean recoveries from spiked samples at three levels ranged from 61 . 9% to 119 . 5% with relative standard deviations of 0 . 8%-18 . 6%. Limits of quantification ranged from 0. 5 μg/kg to 25 μg/kg.

Study on Influences of Enzymolysis and Organic Solvent Extraction on Determination of 2 Kinds of β2-Agonist Residues in Sheep Plasma and Urine / 分析化学

Two kinds of β2-agonistresidues in sheep plasma and urine were disposed by enzymolysis and organic solvent extraction pretreatment methods, and UPLC-MS/MS was used for the qualitative and quantitative analysis. Detection results were compared to study the influences of two pretreatment methods. The experimental results showed that more than 95% of Ractopamine and 40% of Salbutamol exist in the conjugated form in sheep plasma. The detection results of 2 kinds of β2-agonist residues were significantly enhanced when adding β-glucuronidase/aryl sulfatase. The experimental repeatability is very poor ( RSD>40%) when the enzymolysis was not carried out. There were 57% of Ractopamine and less than 1% of Salbutamol exists in the conjugated form in sheep urine. Enzymolysis pretreatment method was useful for the Ractopamine residues determination in urine, and Enzymolysis pretreatment method was useless for Salbutamol determination in urine. Matrix effect of plasma was less than the effects of urine. The influence of organic solvent extraction pretreatment method on the detection results was unremarkable, and there was the possibility that organic solvent extraction could lead partial loss of target compound in extraction process. However, it did not influence the detection results by using internal standard calibration.