OBJECTIVE To establish a method for simultaneous determination of 7 anti-tumor drugs （irinotecan， capecitabine， paclitaxel， docetaxel， tamoxifen， letrozole and methotrexate） in human plasma and apply it to the clinic. METHODS After precipitating with a methanol-acetonitrile mixture （1∶ 1， V/V） containing 0.1% formic acid， liquid chromatography-tandem mass spectrometry （LC-MS/MS） was used to determine the plasma concentration， using deuterium isotopes of each analyte as internal standards. The chromatography was performed on the Agilent Eclipse Plus C18 column with a gradient elution of water （containing 0.1% formic acid+0.04% 5 mmol/L ammonium formate） as mobile phase A and acetonitrile （containing 0.1% formic acid） as mobile phase B. The flow rate was 0.6 mL/min， and the column temperature was set at 40 ℃ . The sample size was 10 μL， and the analysis lasted for 5.5 min. Electrospray ionization was used in positive and negative ion mode， and multiple reaction monitoring mode was used. The ion pairs used for quantitative analysis were m/z 587.1→167.1 （irinotecan）， m/z 360.1→244.1 （capecitabine）， m/z 876.4→308.0 （paclitaxel）， m/z 830.3→304.2 （docetaxel）， m/z 372.1→129.1 （tamoxifen）， m/z 284.1→242.1 （letrozole）， and m/z 455.0→ 308.0 （methotrexate）. A total of 97 patients with malignant tumors in our hospital were selected to measure the plasma concentrations of 7 anti-tumor drugs using the above method. RESULTS The linear ranges of irinotecan， capecitabine， paclitaxel， docetaxel， tamoxifen， letrozole and methotrexate were 2-1 000 ng/mL （r＝0.994 3）， 20-10 000 ng/mL （r＝0.997 5）， 2-1 000 ng/mL （r＝0.997 9）， 1-500 ng/mL （r＝0.995 8）， 1-500 ng/mL （r＝0.995 2）， 1-500 ng/mL （r＝0.996 4）， 10-5 000 （r＝0.997 7）， respectively. The quantitative lower limits were 2， 20， 2， 1， 1， 1 and 10 ng/mL； RSDs of intra-assay precision were 0.08%-14.86% （n＝6）. RSDs of inter-batch precision were 1.51%-11.55% （n＝3）， and the accuracies were 89.17%-114.93% （n＝6）. The matrix effects ranged from 89.89%-119.74% （n＝6）. RSDs of the stability tests were 1.98%-14.88% （n＝6）. The results of E-mail：hangpengzhou@163.com clinical application showed， the average plasma concentrations of irinotecan， capecitabine， paclitaxel and docetaxel were 704.09， 909.40， 36.45， 150.43 ng/mL， respectively. The values of the coefficient of variation were 25.24%， 62.65%， 122.69%， and 92.27%. CONCLUSIONS The established LC-MS/MS method is simple and rapid， and can be used for the simultaneous determination of 7 commonly used anti-tumor drugs in the plasma of patients with malignancy.

A 20-year-old male patient presented to the Department of Dermatology of Peking Union Medical College Hospital with complaints of an 8-year history of facial scarring, swelling of the lower limbs, and a 4-year history of scalp thickening. Physical examination showed thickening furrowing wrinkling of the skin on the face and behind the ears, ciliary body hirsutism, blepharoptosis, and cutis verticis gyrate. Both lower limbs were swollen, especially the knees and ankles. The skin of the palms and soles of the feet was keratinized and thickened. Laboratory examination using bone and joint X-ray showed periostosis of the proximal middle phalanges and metacarpals of both hands, distal ulna and radius, tibia and fibula, distal femurs, and metatarsals.Genetic testing revealed two variants in SLCO2A1, c.1658T > A and c.96+4A > C.Through multidisciplinary consultation, we confirmed the diagnosis of pachydermoperiostosis.

ObjectiveTo observe clinical effectiveness and safety of lumbar oscillatory manipulation in the treatment of patients with lumbar disc herniation. MethodsThis study recruited 90 lumbar disc herniation patients who were randomly divided into 45 cases each in the treatment group and the control group. The treatment group was given lumbar oscillatory manipulation， while the control group was given traditional lumbar obliquely pulling manipulation， and both groups were treated twice a week for 4 weeks. The Oswestry dysfunction index （ODI） score， lumbar spine mobility， and the absolute values of difference in hardness between the left and right sides of the lumbar erector spinae muscle were compared between groups of patients before treatment and after 2 and 4 weeks of treatment； the shear wave elastic modulus of multifidus muscle and abdominal muscle thickness were compared between groups before treatment and after 4 weeks of treatment； the clinical effectiveness of the treatment was compared， and the safety evaluation was carried out. ResultsCompared with the group before treatment， the ODI scores and the the absolute values of difference in hardness between the left and right sides of the lumbar erector spinae muscle decreased after 2 and 4 weeks treatment in both groups， and the mobility of the lumbar vertebrae in all directions of anterior flexion， posterior extension， left lateral flexion， and right lateral flexion increased； the shear wave elastic modulus of bilateral multifidus muscles of the fourth lumbar vertebral segments decreased， and the thickness of abdominal muscle increased after 4 weeks of treatment in both groups （P＜0.05）. Compared with the control group， the ODI score of the treatment group decreased after 2 weeks of treatment， and the shear wave elastic modulus value of bilateral multifidus muscle of the fourth lumbar vertebral segment decreased and the thickness of abdominal muscle increased after 4 weeks of treatment （P＜0.05）. The total effective rate was 90.48% （38/42） in the treatment group and 71.43% （30/42） in the control group， and the clinical effectiveness of the treatment group was better than that of the control group （P＜0.05）. No adverse events occurred in both groups during the study. ConclusionLumbar oscillatory manipulation is superior to traditional lumbar obliquely pulling manipulation in relieving symptoms of lumbar disc herniation and improving lumbar spine function， which may be related to its improvement in the elasticity of the multifidus muscle.

OBJECTIVE: To explore the molecular mechanism of Shenmai Injection (SMI) against doxorubicin (DOX) induced cardiomyocyte apoptosis. METHODS: A total of 40 specific pathogen-free (SPF) male Sprague Dawley (SD) male rats were divided into 5 groups based on the random number table, including the control group, the model group, miR-30a agomir group, SMI low-dose (SMI-L) group, and SMI high-dose (SMI-H) group, with 8 rats in each group. Except for the control group, the rats were injected weekly with DOX (2 mg/kg) in the tail vein for 4 weeks to induce myocardial injury, and were given different regimens of continuous intervention for 2 weeks. Cardiac function was detected by echocardiography and myocardial pathological changes were observed by Van Gieson (VG) staining. Myocardial injury serum markers, including creatine kinase (CK), lactate dehydrogenase (LDH), troponin T (cTnT), N-terminal pro-brain natriuretic peptide (NT-proBNP), soluble ST2 (sST2), and growth differentiation factor-15 (GDF-15) were detected by enzyme linked immunosorbent assay (ELISA). Cardiomyocyte apoptosis was observed by terminal deoxynucleotidyl transferase-mediated biotinylated dUTP triphosphate nick end labeling (TUNEL) and transmission electron microscopy, and the expressions of target proteins and mRNA were detected by Western blot and quantitative real time polymerase chain reaction (qRT-RCR), respectively. RESULTS: The treatment with different doses of SMI reduced rat heart mass index and left ventricular mass index (P<0.05), significantly improved the left ventricular ejection fraction (P<0.05), decreased the levels of serum CK, LDH, cTnT, and NT-proBNP (P<0.05 or P<0.01), reduced the levels of serum sST2 and GDF-15 (P<0.05 or P<0.01), decreased the collagen volume fraction, reduced the expressions of rat myocardial type I and type III collagen (P<0.05 or P<0.01), and effectively alleviated myocardial fibrosis. And the study found that SMI promoted the expression levels of miR-30a and Bcl-2 in myocardium, and down-regulated the expression of Bax, which inhibited the activation of Caspase-3 and Caspase-9 (P<0.05 or P<0.01), and improved myocardial cell apoptosis. CONCLUSIONS SMI can alleviate myocardial injury and apoptosis caused by DOX, and its mechanism possibly by promoting the targeted expression of myocardial Bcl-2 protein through miR-30a.
Animals ; Myocytes, Cardiac/metabolism* ; Apoptosis/drug effects* ; MicroRNAs/genetics* ; Rats, Sprague-Dawley ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Doxorubicin/pharmacology* ; Proto-Oncogene Proteins c-bcl-2/genetics* ; Drug Combinations ; Injections ; Rats

OBJECTIVE: To compare the therapeutic efficacy of portal and tail vein transplantation of bone marrow-derived mesenchymal stem cells (BMSCs) against cholestatic liver fibrosis in mice. METHODS: BMSCs were isolated and co-cultured with starvation-activated hepatic stellate cells (HSCs). HSC activation markers were identified using immunofluorescence and qRT-PCR. BMSCs were injected into the liver tissues of bile duct ligation (BDL) mice via the tail and portal veins. Histomorphology, liver function, inflammatory cytokines, and the expression of key proteins were all determined in the liver tissues. RESULTS: BMSCs inhibited HSC activation by reducing α-SMA and collagen I expression. Compared to tail vein injection, DIL-labeled BMSCs injected through the portal vein maintained a high homing rate in the liver. Moreover, BMSCs transplanted through the portal vein resulted in greater improvement in liver color, hardness, and gallbladder size than did those transplanted through the tail vein. Furthermore, BMSCs injected by portal vein, but not tail vein, markedly ameliorated liver function, reduced the secretion of inflammatory cytokines, including TNF-α, IL-6, and IL-1β, and decreased α-SMA + hepatic stellate cell (HSC) activation and collagen fiber formation. CONCLUSION The therapeutic effect of BMSCs on cholestatic liver fibrosis in mice via portal vein transplantation was superior to that of tail vein transplantation. This comparative study provides reference information for further BMSC studies focused on clinical cholestatic liver diseases.
Animals ; Mice ; Mesenchymal Stem Cell Transplantation ; Liver Cirrhosis/etiology* ; Male ; Cholestasis/therapy* ; Mice, Inbred C57BL ; Hepatic Stellate Cells ; Mesenchymal Stem Cells

OBJECTIVE To investigate whether the microRNA-222(miRNA-222) carried by plasma exosomes can serve as an early warning marker for cognitive impairment induced by shRNA-PCSK9. METHODS The high-fat diet(HFD) was used to prepare a hypercholesterolemic mouse model group. The model group mice were divided into HFD-shRNA control group and HFD-shRNA-PCSK9 group. The shRNA-PCSK9 was constructed, injected intravenously into the body, and the expression of PCSK9 mRNA was detected by real-time PCR(RT-PCR). Tau protein and phosphorylation in brain tissue were observed by immunohistochemistry (IHC). Western blotting was used to detect Tau protein and P-Tau protein. Serum amyloid Aβ1-42Ab levels were determined by ELISA. The kits extracted plasma exosomes step by step, identify the exosome morphology by negative staining electron microscopy, and determined the size of exosomes by NTA technology. RT-PCR technique was used to detect the expression level of miRNA-222 carried in plasma exosomes. RESULTS The model mouse were prepared by feeding HFD for 13 weeks, whose total cholesterol(TC) and low-density lipoprotein(LDL-C) contents in serum were significantly increased. At the same time, the expression of PCSK9 mRNA in the brain tissue of model group was significantly increased. After shRNA-PCSK9 lentivirus interference, PCSK9 mRNA expression was inhibited, and IHC observed that shRNA-PCSK9 induced abnormal expression and hyperphosphorylation of Tau protein in brain tissue, indicating that the pathological changes of neurofibrillary tangles had occurred. However, at this time, serum Aβ1-42Ab had not been significantly increased, and it had not yet been of significance for the diagnosis of cognitive impairment. The miRNA in plasma exosomes was extracted, and RT-PCR results showed that the expression of miRNA-222 carried in the exosomes of the HFD-shRNA-PCSK9 group was significantly lower than that of the HFD-shRNA control group. CONCLUSION Plasma exosomes carried miRNA-222 provides an early warning marker for shRNA-PCSK9- induced cognitive impairment.

OBJECTIVE To optimize the preparation process of sinomenine microemulsion and evaluate its pharmacodynamics.METHODS HPLC method for sinomenine content determination was established,and sinomenine microemulsion prescription was in-itially screened by solubility test and pseudo-ternary phase diagram.D-optimal mixing experimental design method was used to optimize sinomenine microemulsion prescription with particle size and drug load as investigation indexes,and its particle size,drug load and sta-bility were evaluated.Transdermal absorption was investigated by transdermal test in vitro,and the anti-inflammatory effect was evalua-ted by ear swelling test.RESULTS With methanol:0.1%phosphoric acid(40 ∶ 60)as the mobile phase,the detection wavelength was 262 nm,and the method was suitable for the determination of sinomenine.The optimal formula of microemulsion was obtained as castor oil(7.0%),PEG40 hydrogenated castor oil/anhydrous ethanol(69.0%),the optimal Km value was 3∶1,and distilled water(24.0%).The average particle size of the microemulsion was 18.76 nm,the PDI was 0.072 and the drug loading was 5.225%.The cumulative permeability of 1.0%sinomenine microemulsion at 12 h was 1.223 4 μg·cm-2,and the steady permeability rate was 0.0649 μg·cm-2·h-1,which was better than sinomenine solution.The inhibitory rate of sinomenine microemulsion was 65.07%,which was similar to dexamethasone.CONCLUSION The preparation of sinomenine microemulsion has the advantages of stable process,high drug loading,good transdermal absorption and anti-inflammatory effect,which can provide reference for the development of sinomenine transdermal drug delivery preparation.

【Objective】 To analyze the application value of penile vibrating perception threshold measurement in the diagnosis of erectile dysfunction (ED) and provide reference for the seversity of ED patients. 【Methods】 The clinical data, Erectile Hardness Scale (EHS) score, International Index of Erectile Function Questionnaire-5 (IIEF-5) score, and the vibrating perception threshold (VPT) of the penis of 257 patients with decreased erectile function as the main complaint or accompanying symptoms treated during Jan. and Dec.2021 were retrospectively collected and analyzed.Patients with EHS=4 and IIEF-5≥22 were classified into the normal group, and the rest into the ED group.The differences in VPT in patients with different EHS scores were compared, and the correlation between IIEF-5 and VPT was analyzed.The diagnostic value of VPT for ED was evaluated with receiver operating characteristic (ROC) curve. 【Results】 The difference in penile VPT among patients with different EHS scores was statistically significant (P<0.05).With the decrease of EHS score, VPT showed an increasing trend.Glans VPT was negatively correlated with IIEF-5 score (ρ=-0.22, P<0.001), and penile shaft VPT was also negatively correlated with IIEF-5 score (ρ=-0.26, P<0.001).The VPT of glans penis [(4.17±1.37) V vs.(3.47±1.24) V, P=0.009] and the VPT of penis body [(3.73±1.41) V vs.(2.99±1.14)V, P=0.003] in the ED group were both higher than those in the normal group.The area under the ROC curve (AUC) of the glans VPT was 0.642.When the cut-off value was 3.537 V, the sensitivity was 63.4%, and the specificity was 63.6%.The AUC of the penile shaft VPT was 0.659.When the cut-off value was 2.775 V, the sensitivity was 72.3%, with a specificity of 54.5%. 【Conclusion】 The penile VPT of ED patients is higher than that of normal ones, and there is a correlation between VPT and the severity of ED.Severe ED is associated with higher VPT.The measurement of penile VPT is helpful for the clinical diagnosis of ED patients.

Objective:To analyze the treatment efficacy, safety and dose parameters of optimized hippocampus-avoidance prophylactic cranial irradiation (HA-PCI) in limited-stage small cell lung cancer (LS-SCLC) and explore the corresponding dosimetric parameters under the condition of narrowing the hippocampus avoidance region as hippocampus region plus 2 mm in three dimensions.Methods:Clinical data of patients with LS-SCLC receiving HA-PCI (hippocampus avoidance region defined as hippocampus region plus 2 mm in three dimensions) in Cancer Hospital Chinese Academy of Medical Sciences from August 2014 to June 2020 were retrospectively analyzed. Dose parameters of HA-PCI and adverse events were analyzed using descriptive statistics analysis. Changes of neurocognitive function, such as mini-mental state examination (MMSE) and Hopkins verbal learning test-revised (HVLT-R) scores, were evaluated by analysis of variance and Kruskal-Wallis H test. Overall survival (OS), progression-free survival (PFS) and intracranial PFS (iPFS) were calculated using Kaplan-Meier method. The cumulative incidence of local-regional recurrence (LRR), extracranial distant metastases (EDM), and locoregional recurrence (LR) were investigated under competing risk analysis. Results:A total of 112 patients were included, the median follow-up time was 50 months (95% CI: 45.61-54.38). The median volume of hippocampus was 4.85 ml (range: 2.65-8.34 ml), with the average dose ≤9 Gy in 106 patients (94.6%), ≤8 Gy in 92 patients (82.1%). The median volume of hippocampus avoidance area was 15.00 ml (range: 8.61-28.06 ml), with the average dose ≤12 Gy in 109 patients (97.3%), ≤10 Gy in 101 patients (90.2%). The 2-year cumulative LRR, EDM, LR rates were 16.9%, 23.2% and 28.5%, respectively. The 5-year cumulative LRR, EDM, LR rates were 23.2%, 26.9% and 33.3%, respectively. The 2-year iPFS, PFS and OS rates were 66.1% (95% CI: 57.9%-75.4%), 53.6% (95% CI: 45.1%-63.7%) and 80.4% (95% CI: 73.3%-88.1%), respectively. The most common grade I-Ⅱ adverse events were nausea (33.9%) and dizziness (31.3%), and only 1 patient developed grade Ⅲ nausea and dizziness. MMSE ( n=57) and HVLT-R tests ( n=56) showed no significant decline. Conclusions:Optimized HA-PCI can achieve similar dose limitation with favorable efficacy and light toxicity. No significant decline is observed in short-term neurocognitive function in evaluable patients.

BACKGROUND:Ligaments are important structures in maintaining the stability of the lumbar spine,and these structures are prone to degradation due to the generated mechanical stress.However,there are few studies on ligament injuries. OBJECTIVE:To determine the range of motion and stress of each ligament in the state of motion based on a three-dimensional finite element model,and to systematically explore the influence of ligament injury on the stability of the lumbar spine and its biomechanical significance. METHODS:The L4-L5 lumbar finite element model was established.All free forces on the lower surface of L5 were constrained,and a torque of 5 N·m was applied to simulate the motion states of the lumbar spine.Progressive ligament damage was simulated by changing Young's modulus of the ligament. RESULTS AND CONCLUSION:(1)The extension range of motion of most ligament injuries increased significantly.In contrast,there was no significant change in the lateral bending range of motion.(2)The range of motion of capsular ligament injury increased significantly in flexion,extension and lateral bending.The extension range of motion increased significantly after the anterior longitudinal ligament injury.Intertransverse ligament injury resulted in a significant increase in the lateral bending range of motion.(3)After a single ligament injury,the most significant change in range of motion was observed during extension.After a single ligament injury,the stress of the remaining ligaments increased,especially the capsular ligament.The stress changes of the interspinous ligament and ligamentum flavum were the least obvious.Ligament stress changes least in lateral bending and most in torsion.(4)Ligament damage did not result in major changes in disc pressure,indicating that ligament injury leads to an increased range of motion of the lumbar spine and affects the stability of the lumbar spine.(5)Capsular ligament was stable in flexion,extension and lateral bending.The anterior longitudinal ligament showed a stable extension;the intertransverse ligament stabilizes the lateral curve.(6)Extension exercise is sensitive to a ligament injury,and the pathological changes of the ligament can be examined by extension exercise.(7)Stress compensation was given to the remaining ligaments to maintain the stability of the lumbar spine after a single ligament injury.(8)Interspinous ligament and ligamentum flavum injuries have the least impact on the peripheral ligaments,while capsular ligament injuries have the greatest impact on the peripheral ligaments.(9)The ligament injury has the least effect on the residual ligament stress during lateral bending exercise,while it has the greatest effect on the ligament stress during the twisting exercise.Patients with ligament injury should avoid twisting exercises.Ligament injuries do not affect disc pressure.