A strategy combining a tailored database and high-throughput activity screening that discover bioactive metabolites derived from Magnoliae Officinalis Cortex (MOC) was developed and implemented to rapidly profile and discover bioactive metabolites in vivo derived from traditional Chinese medicine (TCM). The strategy possessed four characteristics: 1) The tailored database consisted of metabolites derived from big data-originated reference compound, metabolites predicted in silico, and MOC chemical profile-based pseudomolecular ions. 2) When profiling MOC-derived metabolites in vivo, attentions were paid not only to prototypes of MOC compounds and metabolites directly derived from MOC compounds, as reported by most papers, but also to isomerized metabolites and the degradation products of MOC compounds as well as their derived metabolites. 3) Metabolite traceability was performed, especially to distinguish isomeric prototypes-derived metabolites, prototypes of MOC compounds as well as phase I metabolites derived from other MOC compounds. 4) Molecular docking was utilized for high-throughput activity screening and molecular dynamic simulation as well as zebrafish model were used for verification. Using this strategy, 134 metabolites were swiftly characterized after the oral administration of MOC to rats, and several metabolites were reported for the first time. Furthermore, 17 potential active metabolites were discovered by targeting the motilin, dopamine D2, and the serotonin type 4 (5-HT₄) receptors, and part bioactivities were verified using molecular dynamic simulation and a zebrafish constipation model. This study extends the application of mass spectrometry (MS) to rapidly profile TCM-derived metabolites in vivo, which will help pharmacologists rapidly discover potent metabolites from a complex matrix.

ObjectiveTo systematically and objectively characterize the pharmacological effects of Fufang Biejia Ruangan pills （FBRP） in the intervention of alcoholic liver disease （ALD） using acute and chronic ALD mouse models and to elucidate its molecular mechanisms. MethodFifty SPF-grade male BALB/c mice were randomly divided into the normal group， model group， and FBRP low-， medium-， and high-dose groups （9.6， 19.2， 38.4 mg·kg^-1）. Except for the normal group， the remaining groups were given 56° white wine by gavage to establish the acute ALD model， with samples collected after 4 weeks. Thirty SPF-grade male C57BL/6N mice were randomly divided into the normal group， model group， and FBRP medium-dose group （19.2 mg·kg^-1）. The chronic ALD mouse model was established using the Lieber-DeCarli method over a 10-week period. Inflammatory markers in liver tissues were assessed using hematoxylin-eosin （HE）， Sirius Red， oil red O staining， and enzyme-linked immunosorbent assay （ELISA）. Intoxication behaviors of each group were objectively evaluated through sobering-up time， net-catching， and pole-climbing tests. Further bioinformatics analyses based on clinical transcriptomic data were conducted to identify key targets and molecular mechanisms of FBRP in alleviating ALD through liver-brain dialogue， with experimental validation by ELISA， Western blot， and immunohistochemical staining. ResultCompared with the normal group, the levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in liver tissues of mice in the acute and chronic ALD model groups were significantly increased （P<0.05）. Compared with the model group， the levels of AST and ALT in liver tissue of mice in FBRP groups were significantly decreased （P<0.05）. Compared with the normal group， the time of grasping the net and climbing the pole in the acute ALD model group was significantly decreased within 4 weeks （P<0.01）. Compared with the model group， the grasping and climbing time of FBRP high dose groups increased significantly within 4 weeks （P<0.05）. Compared with the normal group， the expression of high mobility group protein B1 （HMGB1） protein in liver tissue and prefrontal lobe tissue of mice in the chronic ALD model group was significantly increased （P<0.01）. Compared with the model group， the expression of HMGB1 protein in FBRP medium dose group was significantly decreased （P<0.05，P<0.01）. Compared with the normal group， the expression of brain-derived neurotrophic factor （BDNF） protein and the release of γ-aminobutyric acid （GABA） in the prefrontal cortex of the model group were significantly decreased （P<0.01）. Compared with the model group, the expression of BDNF protein and the release of GABA in the FBRP medium dose group were significantly increased （P<0.05）. ConclusionThis study revealed that FBRP improved key pathological changes in ALD by modulating liver-brain dialogue mediated by the HMGB1-BDNF axis. These findings provide experimental evidence for the clinical use of FBRP in treating ALD and offer new insights for the development of ALD therapeutic agents.

ObjectiveTo identify the pharmacodynamic material basis of Ruyi Zhenbaowan in relieving neuropathic pain by integrating the calculation of biological network proximity and pharmacokinetic characterization. MethodThe interaction network of "drug candidate target-related gene of disease" was constructed by Cytoscape 3.8.2， and the average shortest path value of each drug putative target acting on neuropathic pain-related genes in this network was calculated by Pesca 3.8.0 tool so as to evaluate the network proximity between them， and screen prescription candidate targets with strong intervention efficiency and their corresponding potential effect components. After that， plasma and cerebrospinal fluid samples were collected from rats after administration of Ruyi Zhenbaowan at set time points， and the contents of potential effect components in samples was quantified by ultra performance liquid chromatography-quadrupole-ion trap mass spectrometry（UPLC-Q-TRAP/MS）， and drug concentration-time curves were plotted， then the pharmacokinetic parameters were calculated by DAS 2.1.1. ResultBy evaluating the network proximity between candidate targets and neuropathic pain-related genes in the interaction network， a total of 40 putative targets of Ruyi Zhenbaowan with strong intervention effects on neuropathic pain-related genes， such as estrogen receptor 1（ESR1）， cyclic adenosine monophosphate（cAMP）-dependent protein kinase catalytic subunit alpha（PRKACA） and protein kinase B1 （Akt1）， and 10 corresponding potential effect components， such as glycyrrhizic acid and betulinic acid， were obtained. Pharmacokinetic characterization showed that among the 10 potential effect components， gallic acid， apigenin-7-O-glucuronide， glycyrrhizic acid and apigenin were well absorbed and metabolized in plasma and cerebrospinal fluid， with long onset time and good bioavailability. ConclusionFrom the perspective of efficacy-target-constituent-pharmacokinetic， this study analyzes the main effective materials of Ruyi Zhenbaowan， such as glycyrrhizic acid， gallic acid， apigenin-7-O-glucuronide and apigenin， which have a high exposure in plasma or cerebrospinal fluid and have a strong intervention effect on neuropathic pain. The related results provide reliable experimental evidences for clarifying the material basis and developing quality standards of Ruyi Zhenbaowan.

ObjectiveTo identify the chemical constituents of Ruyi Zhenbaowan in vitro and in vivo. MethodThe chemical constituents of Ruyi Zhenbaowan were identified based on UHPLC-Q Exactive Orbitrap HRMS. A total of 12 male SD rats were randomized into two groups： control （pure water） and Ruyi Zhenbaowan （1.8 g·kg^-1）. The rats were administrated with the suspension of Ruyi Zhenbaowan or pure water by gavage. After 1.5 h， the plasma and cerebrospinal fluid were collected. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C₁₈ column （2.1 mm × 150 mm， 1.7 μm） with a mixture of 0.1% formic acid aqueous solution （A） and acetonitrile （B） as the mobile phase. Gradient elution was carried out according to the procedure of 0~15 min，97%~80%A；15~30 min ，80%~60%A；30~40 min，60%~30%A；40~45 min，30%~5%A. The ion source was electrospray ionization， and scan range was m/z 100-1 500. The prototype components and the components in the plasma and cerebrospinal fluid were analyzed qualitatively by scanning in positive and negative ion modes and identified by comparison with the data in published literature and the information of standard substances. ResultA total of 126 chemical constituents were identified from the 80% methanol solution of Ruyi Zhenbaowan， and 14 and 7 prototype constituents were detected in the plasma and the cerebrospinal fluid， respectively. In addition， the fragmentation rules of apigenin， apigenin-7-O-glucuronide， galangin， liquiritin， piperine， glycyrrhizic acid， eugenol， gallic acid， and cholic acid were deduced. ConclusionThis study achieved rapid multicomponent characterization and identification of Ruyi Zhenbaowan in vitro and in vivo， providing theoretical support for exploring active substances and performing quality control.l.

Objective:To analyze the clinical characteristics of gastrointestinal symptoms and liver function injury in patients with coronavirus disease 2019 (COVID-19).Methods:From January 23, 2020 to February 29, 2020, the medical records of 251 patients with COVID-19 admitted to the West Campus of the Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, were collected. The proportion of the patients with gastrointestinal symptoms including anorexia, nausea and vomiting, diarrhea and abdominal pain were analyzed respectively. The patients were divided into common type (76 cases), severe type (65 cases) and critical type (110 cases). The incidence of liver function injury and the changes of liver function parameters such as total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH), albumin and globulin of the patients with different clinical types and with or without gastrointestinal symptoms were analyzed. Mann-Whitney U test, Chi square test and Fisher′s exact test were used for statistical analysis. Results:The main gastrointestinal symptoms of patients with COVID-19 were anorexia (33.9%, 85/251), diarrhea (12.0%, 30/251), nausea and vomiting (7.6%, 19/251) and abdominal pain (1.2%, 3/251). 143 patients (57.0%) had liver function injury, the rate of liver function injury in critical type patients was 75.5% (83/110), which was higher than that of common type patients (40.8%, 31/76) and severe type patients (44.6%, 29/65), and the differences were statistically significant ( χ2=22.765 and 16.865, both P<0.01). There was no significant difference in the proportion of patients with liver function injury between common type and severe type patients ( P>0.05). There was no statistically significant difference in the proportion of liver function injury between patients with gastrointestinal symptoms and those without gastrointestinal symptoms (57.8%(67/116) vs. 56.3%(76/135), P>0.05). The median values of TBil, DBil, ALT, AST, ALP, GGT, LDH and globulin level of critical type patients were 13.5 μmol/L, 4.9 μmol/L, 44.5 U/L, 50.0 U/L, 64.0 U/L, 41.0 U/L, 527.0 U/L and 33.6 g/L respectively. The proportions of critical type patients with TBil level >34.2 μmol/L, DBil level>13.6 μmol/L, ALT level>80 U/L and AST level>80 U/L were 7.3% (8/110), 7.3% (8/110), 17.3% (19/110) and 17.3% (19/110), respectively. These results were all higher than those of common type patients (9.5 μmol/L, 2.9 μmol/L, 28.5 U/L, 28.5 U/L, 54.0 U/L, 25.5 U/L, 225.5 U/L, 30.1 g/L, 0, 0, 6.6% (5/76) and 2.6% (2/76) ) and severe type patients (10.4 μmol/L, 3.4 μmol/L, 30.0 U/L, 31.0 U/L, 49.0 U/L, 25.0 U/L, 284.0 U/L, 30.7 g/L, 0, 0, 6.2% (4/65) and 1.5% (1/65)), and the differences were statistically significant ( Z=-4.264, -5.507, -4.000, -6.558, -3.112, -4.333, -4.858, -3.873, Fisher′s exact test, Fisher′s exact test, χ2=4.574, 9.620; Z=-3.060, -3.850, -3.923, -5.005, -9.495, -7.651, -3.853, -2.725, Fisher′s exact test, Fisher′s exact test, χ2=4.425, 10.169; all P<0.01). The median values of pre-albumin level, albumin level and the albumin to globulin ratio of critical type patients were 85.3 g/L, 28.2 g/L and 0.8, which were all lower than those of common type patients (157.3 g/L, 32.3 g/L and 1.1, respectively) and severe type patients (133.6 g/L, 31.6 g/L and 1.1, respectively), and the differences were statistically significant ( Z=-6.631, -3.647, -4.924, -4.503, -5.283 and -3.903, all P<0.01). The median albumin level of patients with diarrhea was lower than that of patients without diarrhea (28.2 g/L vs. 30.5 g/L), the proportion of diarrhea patients whose TBil level >20.0 to 34.2 μmol/L was higher than that of patients without diarrhea (70.0%, 21/30 vs. 10.9%, 24/221), and the differences were statistically significant ( Z=-2.182, χ2 =62.788; both P<0.05). Conclusions:Anorexia is the most common digestive symptom in COVID-19 patients, and the incidences of abdominal pain is low. The incidence of liver function injury of critical type patients is high. There is no significant correlation between gastrointestinal symptoms and liver function injury, and patients with diarrhea have lower albumin levels.

Objective To establish a laparoscopic classification of extrahepatic biliary dilatations (EHBD) that can guide minimally invasive treatment.Methods According to inclusion criteria,124 patients with EHBD who were admitted and treated from July 2001 to July 2017 in the First Hospital Affiliated to Army Military Medical University were included in this study.A new laparoscopic classification of EHBD was proposed based on the preoperative imaging data and laparoscopic findings of the position and extent of EHBD.The minimally invasive diagnosis and treatment strategies were made based on the new classification.Results According to the preoperative imaging data and intraoperative laparoscopic findings,124 patients with EHBD were divided into the following groups:type A (upper segment,34 cases),type B (middle segment,27 cases),type C (lower segment,20 cases),and type D (entire bile duct,43 cases).The clinical symptoms (abdominal pain,jaundice and mass) and reoperation rates were not significantly different among the 4 groups(both P＞0.05).The incidences of comorbidities (calculus or inflammation) were significantly different (P＜0.05).The operative time(type A:237.6±66.7 min,type B:259.2±60.0 min,type C:286.1 ± 74.7 min,type D:347.5±94.4 min) and blood loss (type A:192.6±102.2 ml,type B:201.5±120.2 ml,type C:297.5±162.1 ml,type D:305.8±237.3 ml) were significantly different among the groups (P＜ 0.05).The short-term complication rates after surgery (5.9％ ～ 20.0％) were significantly different (P＜ 0.05),while the long-term complication rates after surgery (7.4％ ～ 10.0％) were not significantly different.The conversion rates to open surgery were significantly higher in patients with type C and D lesions than in those with type A and B lesions (P＜0.05).Conclusion This laparoscopic classification predicted the difficulty of laparoscopic surgery for EHBD and had a guiding significance in the minimally invasive treatment for this disease entity.

BACKGROUND/AIMS: Currently, there exists no biomarker for visceral hypersensitivity in irritable bowel syndrome (IBS). Piezo proteins have been proven to play an important role in the mechanical stimulation to induce visceral pain in other tissues and may also be a biomarker candidate. The aim of this study was to test the expressions of Piezo1 and Piezo2 proteins in the intestinal epithelial cells from different intestinal segments and to explore the correlation between Piezo proteins expression and visceral pain threshold. METHODS: Post-infectious IBS was induced in mice via a Trichinella spiralis infection. Visceral sensitivity was measured with abdominal withdrawal reflex to colorectal distention. Inflammation in the small intestine and colon was scored with H&E staining. Expression location of Piezo proteins was confirmed by immunohistochemistry. Abundance of Piezo proteins were measured with real-time reverse transcriptase polymerase chain reaction. RESULTS: Piezo1 and Piezo2 proteins were expressed in the intestinal epithelial cells. The expression levels of Piezo1 and Piezo2 were abundant in the colon than the small intestine (P < 0.001 for Piezo1, P = 0.003 for Piezo2). Expression of Piezo2 in the colon significantly correlated to the visceral sensitivity (r = −0.718, P = 0.001) rather than the mucosal inflammation. CONCLUSION: Piezo2 is a candidate biomarker for visceral hypersensitivity in IBS.
Animals ; Colon ; Epithelial Cells ; Humans ; Hyperalgesia ; Hypersensitivity* ; Immunohistochemistry ; Inflammation ; Intestine, Small ; Ion Channels ; Irritable Bowel Syndrome* ; Mice ; Reflex ; Reverse Transcriptase Polymerase Chain Reaction ; Trichinella spiralis ; Visceral Pain

Objective To compare the accuracy of dynamic contrast-enhanced magnetic resonance (DCE-MRI) and SPECT in the measurement of glomerular filtration rate (GFR) in renal allografts.Methods Sixty renal transplant recipients were enrolled in this study.DCE-MRI and SPECT were used to measure the GFR of the transplanted kidneys,and compared with the endogenous creatinine clearance rate (Ccr).Bias,precision,correlation and Bland-Altman agreement were calculated for each modality compared with the endogenous Ccr.Results In 60 renal transplant recipients,the corrected Ccr was (60.63 ± 24.83) ml · min-1 · 1.73 m-2.The GFR measured by SPECT was (65.31 ± 17.08) ml · min-1 · 1.73 m-2,and (50.44 ± 22.78) ml · min-1 · 1.73 m-2 by MRI,respectively.The bias of GFR-SPECT was 4.69 ml·min-1 · 1.73 m-2,and the precision was 23.76 ml·min-1 1.73 m-2.The bias of GFR-MRI was-10.18 ml·min-1 ·1.73 m-2,and the precision was 13.87 ml·min-1 · 1.73 m-2.Correlation analysis showed that GFR-MRI and the endogenous Ccr had a good correlation (r=0.833,P＜0.01),GFR-SPECT and the endogenous Ccr had a moderate correlation (r=0.406,P＜0.01),and GFR-MRI and GFR-MRI had a poor correlation (r=0.342,P ＜0.01).Bland-Altman analysis showed a confidence interval of 95.3 ml·min-1 ·1.73 m-2 for GFR-SPECT and 62.3 ml· min-1 · 1.73 m-2 for GFR-MRI.Conclusion DCE-MRI can be used as confidently as SPECT to evaluate the renal function of transplanted kidneys in the same time of determining anatomical information.

ObjectiveTo investigate the effect of acup point injection and rehabilitation techniques on recovery of upper limb function and activities of daily life (ADL) in patients with shoulder-hand syndrome (SHS) after cerebral infarction.Methods Ninety-six patients with SHS after cerebral infarction admitted to Jiuquan City Traditional Chinese Medicine Hospital and Jiuquan City People's Hospital from January 2014 to February 2016 were enrolled, and they were randomly assigned to observation group (56 cases) and control group (40 cases). The patients in the two groups were given the same routine comprehensive cerebral infarction treatments and rehabilitation technological training. In the control group, additionally intermediate frequency physiotherapy was used, while in the observation group, acupuncture and acupoint injection of salvia miltiorrhiza and ligustrazine were added. The patients in two groups were treated consecutively for one month, afterwards, the upper limb motor function, ADL, shoulder joint pain, degree of swelling and range of motion were assessed, and the clinical effects were observed.Results After treatment, the modified Fugl-Meyer motor function score (FMA), modified barthel index (MBI) and the ranges of voluntary activities of shoulder abduction, adduction, flexion, extension, internal rotation and external rotation in patients of two groups were significantly increased than those before treatment; the pain visual analog scale (VAS) and the edema volume difference in observation group were lower than those in the control group, and after treatment, the changes of above indexes were more significant in the observation group than those in the control group [FMA: 70.10±6.38 vs. 41.76±18.69, VAS: 2.00±1.37 vs. 5.00±1.25, MBI: 75.17±3.51 vs. 51.56±2.12, edema volume difference (cm3): 2.13±2.05 vs. 5.75±1.17, flexion: (153.1±6.7)° vs. (56.0±10.9)°, extension: (39.6±5.8)° vs. (17.2±6.7)°, abduction:(168.1±9.6)° vs. (52.1±8.9)°, adduction: (52.1±3.6)° vs. (21.8±4.3)°, external rotation: (49.8±7.6)° vs. (23.2±5.2)°, internal rotation: (107.0±8.5)° vs. (51.2±6.1)°], the total effect in observation group was significantly higher than that in the control group [91.1% (52/56) vs. 70.0% (28/40),P < 0.05].Conclusions Acupuncture and acupoint injection combined with three-level therapeutic regimen of rehabilitation technological training for treatment of shoulder-hand syndrome after cerebral infarction can reduce the patient's pain and edema, and improve upper limb motor function and activities of daily living.

Objective To analyze the doctors' working enthusiasm and its influencing factors on county level hospitals.Methods 17 seminars were held in 8 counties of 3 provinces to investigate 1055 doctors with questionnaires, analyzing the three scales of working enthusiasm, working enthusiasm influencing factors, and changes since the 2009 new healthcare reform.Results Working enthusiasm comprises inner drive, working pleasure, work recognition and external recognition, and inner drive of which is found to play the most important part, with 20.55％ variance contribution.Among the six dimensions of the influencing factors, working conditions rank the highest with 19.46％ variance contribution.Conclusion Optimized measures in the healthcare reform, better working conditions and social environment can improve working enthusiasm of these doctors.