Objective: To study the chemical components of Citrus medica L.var. sarcodactylis Swingle and to supply evidence for the quality control. Methods: Chromatography and recrystallization methods were applied to analyze the components. Results and Conclusion: Three compounds were extracted from Citrus medica L. var. sarcodactylis (Noot) Swingle. And their structures were identified as Limettin (I), stigmasta-5,22-dien-3-ol (II) and palmitic acid (III) by the methods of UV, MS, IR, 1HNMR, 13CNMR and DEPT. Stigmasta-5,22-dien-3-ol (II) was obtained by extraction.

Objective To study the influence of irradiation on the osteoblast function by the gene expression changes of RANKL and OPG.Methods Bone marrow stromal cells were induced to develop into early and mature osteoblasts in vitro.The characterization of osteoblasts was indentified by ALP staining.The RANKL and OPG mRNA levels in early and mature osteoblasts, which exposed to 0 -4 Gy radiation were determined by RT-PCR.Results Bone marrow stromal cells had been induced to early and mature osteoblasts by osteoblast differentiation medium in vitro.In early stage of osteoblast, RANKL mRNA expression levels treated with 1Gy irradiation was 2.83-fold higher than those other irradiation dosage groups.The RANKL mRNA expression levels of each group in early stage of osteoblasts were significantly higher than those in the mature counterpart ( t = 8.34 - 103.57, P ＜ 0.05 ).The ratio of RANKL/OPG mRNA was obviously greater in early osteoblast compared with the mature cells ( t = 2.84 - 20.99, P ＜0.05 ), and it was the highest in 1Gy irradiation treated early osteoblast.Conclusions Radiation exposure of the early osteoblasts promotes osteoclasts function and results in the bone loss.

Objective To investigate the correlation of miR-181a and miR-181b with fucosyltransferase FUT1, the functional mechanism was elucidated in a colorectal cancer ( CRC).Methods It collected 32 pairs of tissue samples , 18 males and 14 females in the first affiliated hospital of Dalian Medicinal University, from March 2014 to January 2016.The expression of miR-181a and miR-181b was detected by RT-PCR in CRC tissues , adjacent tissues , serum of colorectal cancer patients and healthy people, and CRC cell lines SW620 and SW480 with differently metastatic ability.The relationship of FUT1 and miR-181a, miR-181b expression were verificated by Pearson's correlation curve.FUT1 was identified the target of miR-181a and miR-181b by Network prediction softwares ( TargetScan Human 7.1, microRNA.org and Starbase v2.0) and luciferase assay.The effects of miR-181a and miR-181b expression on the proliferation, migration, invasion and angiogenesis of SW 480 and SW620 cells were further detected by CCK8, wound healing, transwell and tube foramtion assays.T-test was used for comparison between two independent samples , and one-way anova was used for comparison between multiple samples . Pearson correlation coefficient was used for correlation analysis .Results The levels of miR-181a and 181b in CRC tissues were much lower than in tumor-adjacent tissues (3.12 ±1.88 vs 6.44 ±2.32, t=11.74;3.16 ± 1.77 vs 5.52 ±2.45, t=3.24 ;P<0.05).The levels of miR-181a and 181b in serum of colorectal cancer patients were much lower than in healthy people (1.32 ±0.25,2.57 ±0.48,t=10.26;0.91 ±0.14,1.63 ± 0.29,t=5.19;P<0.05 ) .The levels of miR-181a and miR-181b in SW620, SW480 CRC cells were detected to be much lower than in normal colorectal epithelial cells [(0.65 ±0.10, 0.50 ±0.09) vs 1.0;(0.60 ±0.12,0.42 ±0.03)vs 1.0;t=3.08, P<0.05].FUT1 was highly expressed in CRC tissues and SW620 (t=5.23, P<0.05).Based on the network prediction softwares and luciferase assays , FUT1 was the common target of miR-181a and miR-181b.Over expression of miR-181a or miR-181b inhibited FUT1 level and attenuated the capacity of cell migration , invasion and proliferation in SW 620.Down-regulation of miRNAs in SW480 increased FUT1 expression and promoted the capability of cell migration , invasion and proliferation.Downregulation of the two miRNAs attenuated the capability of cell invasion in SW 480, which was blocked by the reductive FUT1.Conclusion MiR-181a and miR-181b mediated the progression of CRC cells by targeting FUT1.

OBJECTIVE: To evaluate the protective effect of low dose ouabain on injury of cultured spiral ganglion neurons evoked by trophic factors deprived and to explore the mechanism. METHOD: Spiral ganglion neurons were cultured in vitro for 7 days, and then exposed to Neurobasal medium + B27 supplement, Neurobasal medium only or Neurobasal medium + 10 nmol/L ouabain, respectively. After 48 h exposed to different medium, spiral ganglion neurons were stained by FITC labeled Annexin-V and PI, then apoptosis index were calculated using fluorescent microscope and flow cytometry, respectively. In addition, spiral ganglion tissues were cultured for 48 h to evaluate dendrite growth of spiral ganglion neurons in each group. Immunocytochemistry were performed to detect the level of Bcl-2 in each group at 6 h and 12 h. RESULT: Spiral ganglion neurons exposed to Neurobasal medium +10 nmol/L ouabain have a similar apoptosis index compare with that of Neurobasal medium + B27 supplement, but a much lower apoptosis index than that of Neurobasal medium only. In addition, dendrite growth of spiral ganglion neurons exposed to Neurobasal medium +10 nmol/L ouabain was much longer than that of Neurobasal medium only. Bcl-2 level increased in spiral ganglion neurons exposed to Neurobasal medium + 10 nmol/L ouabain at 6 h. CONCLUSION Low dose of ouabain protects injury of spiral ganglion neurons evoked by trophic factors deprived in vitro. This effect may mediated by increasing the level of Bcl-2.
Animals ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Ouabain ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Spiral Ganglion ; drug effects ; pathology

Objective To investigate the plasma levels of signal peptide-CUB epidermal growth fac-tor domain inclusion protein-1(SCUBE-1),asymmetric dimethylarginine(ADMA)and insulin-like growth factor binding protein-2(IGFBP-2)in elderly patients with acute pulmonary embol-ism(PE)and their relationship with risk stratification and poor prognosis.Methods A total of 117 elderly patients with acute PE admitted to our hospital from January 2022 to December 2023 were retrospectively enrolled in this study.With risk stratification,there were 41 patients of high-risk,46 of medium-risk and 30 of low-risk.According to their clinical outcomes,they were divided into good prognosis group(85 cases)and bad prognosis group(32 cases).The plasma levels of SCUBE-1,ADMA,and IGFBP-2 were measured using ELISA.Multivariate logistic regression analysis was applied to identify the risk factors for poor prognosis,and ROC curves were drawn to analyze the predictive value of plasma levels of SCUBE-1,ADM A and IGFBP-2 for poor prognosis of these patients.Results The levels of cTnI,BNP,D-dimer,SCUBE-1,ADMA and IGFBP-2 were significantly higher in the poor prognosis group than the good prognosis group(0.95±0.28μg/L vs 0.30±0.08 μg/L,2017.62±308.15 μg/L vs 538.14±102.73 μg/L,5.24±1.13 μg/L vs 1.63±0.27 μg/L,22.50±10.14 μg/L vs 9.25±4.36 μg/L,2.01±0.72 μmol/L vs 0.61±0.17μmol/L,468.16±90.51 μg/L vs 155.27±30.74 μg/L,P＜0.01).The high-risk patients had high-er plasma levels of SCUBE-1,ADMA and IGFBP-2 than the medium-and low-risk patients in or-der(all P＜0.01).Multivariate logistic regression analysis showed that elevated levels of cTnI,BNP,D-dimer,SCUBE-1,ADMA and IGFBP-2 were risk factors for death in acute PE patients(P＜0.05).ROC curve analysis indicated that the AUC value of SCUBE-1,ADMA and IGFBP-2 combined together in predicting death for acute PE patients was the highest(0.927,95％CI:0.864-0.993),with an accuracy of 93.1％.The plasma levels of SCUBE-1,ADMA and IGFBP-2 were positively correlated with the levels of cTnI,BNP and D-dimer in the elderly patients(P＜0.01).Conclusion For the elderly patients with acute PE,elevated plasma levels of SCUBE-1,ADMA and IGFBP-2 are associated with risk stratification and poor prognosis.Combination of these three indicators has certain predictive value for poor prognosis in the patients.

Objective We aimed to establish and evaluate a rat model of post-infectious irritable bowel syndrome(PI-IBS)with the pattern of liver depression and spleen deficiency coupled with dampness.Methods First,200 rats were randomly divided into the normal group,the infection group,the infection+stress group,the infection+stress+external dampness group,and the acetic acid+stress group(n=40 rats per group)for eight weeks.The rats were treated with Trichinella spiralis infection,chronic restraint stress,an artificial high-humidity climate,and/or acetic acid enema.Weight growth rate,24-hour food intake and water intake,and the fecal moisture percentage were recorded.The open field test and the sucrose consumption test were used to determine the behavioral characteristics of rats in each group.The abdominal withdrawal reflex(AWR)test was used to determine visceral sensitivity.The contents of 5-hydroxytryptamine(5-HT)and aquaporin 4(AQP4)in colon tissue were detected by ELISA.Results Compared with the normal group,the weight growth rate of rats in the infection+stress+external dampness group and the acetic acid+stress group was lower from Week 1 to Week 8 of modeling.The 24-hour food intake of rats in the infection+stress+external dampness group and the infection+stress group was lower than that in the normal group from Week 2 to Week 8 of modeling.At the end of week 2 of modeling,the 24-hour water intake of rats in the infection+stress+external dampness group,the acetic acid+stress group,and the infection+stress group was lower than that in the normal group.The fecal moisture percentage of rats in the infection+stress+external dampness group was higher than that in the normal group at the end of Week 1,6,and 8(P＜0.05).At the end of Week 4 of modeling,the total distance in the open field test in the infection+stress+external dampness group and the acetic acid+stress group was shorter than that in the normal group.The sugar preference rate in the infection+stress+external dampness group was lower than that in the normal group at the end of 1,4,and 8 weeks and lower than that in the acetic acid+stress group at the end of Week 4 and 8(P＜0.05).The AWR scores of rats in the infection+stress+external dampness group were higher than those in the normal group after week 1 at 60 and 80 mmHg(1 mmHg≈0.133 kPa),after Week 4 at 40,60,and 80 mmHg,and after Week 6 and 8 at 20,40,and 80 mmHg(P＜0.05).At the end of Week 2 and 4,a large number of inflammatory cells were infiltrated in the colonic mucosa of the intervention groups,and the inflammation score was higher than that of the normal group(P＜0.05).At the end of weeks 6 and 8,the inflammatory cell infiltration in the intestinal mucosa of the intervention groups was not obvious,and the colonic mucosa returned to normal.At the end of weeks 6 and 8,the 5-HT content was higher in the infection+stress group,the infection+stress+external dampness group,and the acetic acid+stress group than in the normal group(P＜0.05).After Week 4,the AQP4 content was lower in the infection+stress+external dampness group and the acetic acid+stress group than that in the normal group(P＜0.05).After week 6,compared to the normal group,the AQP4 content was lower in all groups except for the acetic acid+stress group,and the AQP4 content in the infection+stress+external dampness group was lower than that in the acetic acid+stress group.After week 8,only in the infection+stress+external dampness group the AQP4 content was lower than in the normal group(P＜0.05).Conclusion The combination of Trichinella spiralis infection,chronic restraint stress,and an artificial high-humidity climate can be used to prepare a relatively stable and reliable rat model of PI-IBS with the pattern of liver depression and spleen deficiency with dampness.

【Objective】 To investigate the distribution characteristics of Rh blood group antigen phenotypes, haplotypes and irregular antibodies between patients in our hospital and local blood donors, so as to ensure safe and effective blood transfusion and improve the rationality and scientificity of clinical blood transfusion. 【Methods】 A total of 113 326 blood samples, from hospitalized patients in our hospital and local blood donors from October 2015 to March 2020, were subjected to Rh antigen typing and irregular antibody detection. The frequency of Rh phenotypes, haplotypes, and irregular antibodies were retrospectively analyzed and calculated. Chi square test was used to compare the data among different population groups. Rh antigen typing and irregular antibody detection were completed using the automatic blood group analyzer. 【Results】 The prevalence of negative RhD was 0.36% (408/113 326). The most prevalent Rh phenotype was DCCee [40.69%(46 112/ 113 326)] followed by DCcEe [36.82%(41 727/ 113 326)]. Anti-E was the most common irregular antibody, accounting for [0.26%(295/ 113 326)], and DCe [62.51%(70 840/ 113 326)] was the most common haplotype. The most common Rh phenotypes and haplotypes in Caucasians in Germany, North Indian and North African were DCcee, DCCee and Dccee, while DCe, DCe and Dce, respectively. 【Conclusion】 The distribution characteristics of Rh phenotypes, haplotypes and irregular antibodies of patients in our hospital and local blood donors were in line with the distribution characteristics of the population in northern China. Corresponding plans concerning blood storage and collection, as well as the establishment of Rh blood type registry should be carried to effectively ensure the safety, rationality and accuracy of clinical blood transfusion.

OBJECTIVE To establish the ultra-high performance liquid chromatography （UPLC） fingerprint of Jingtian granule， and to evaluate its quality by chemical pattern recognition. METHODS Luna® Omega Polar C18 column （150 mm×2.1 mm， 1.6 μm） was used as the chromatographic column， and acetonitrile-0.2% phosphoric acid solution was used as the mobile phase for gradient elution. The flow rate was 0.2 mL/min， the column temperature was 30 ℃， and the detection wavelength was 265 nm. With peak 16 as the reference peak， the UPLC fingerprint of Jingtian granule was established by the Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）. The common peaks were identified， the similarity evaluation was carried out， and the ownership of each common peak was confirmed. Hierarchical cluster analysis （HCA） and principal component analysis （PCA） in chemical pattern recognition methods were used to classify 13 batches of samples （S1- S13）， and orthogonal partial least squares-discriminant analysis （OPLS-DA） was used to identify the key components of the differences between different batches of samples. RESULTS RSDs of precision， repeatability and stability of the UPLC method were not more than 4.4%. A total of 25 common peaks were identified in the fingerprints of 13 batches of Jingtian granules. By comparing with the reference substance fingerprint， 10 common peaks were identified， namely peak 3 （hydroxymethyl-2-furaldehyde）， peak 5 （salidroside）， peak 8（chlorogenic acid）， peak 15 （cinnamic acid）， peak 19 （aloe-emodin）， peak 20 （ammonium glycyrrhizinate）， peak 21 （rhein）， peak 23 （emodin）， peak 24 （glycyrrhetinic acid）， peak 25 （chrysophanol）. The similarities of fingerprints of 13 batches of samples were 0.955-0.996. The results of HCA showed that 13 batches of samples could be divided into three categories， among which samples S1， S5， S7， S11-S13 were clustered in one category， S4 and S6 were clustered in one category， S2， S3 and S8-S10 were clustered in one category. PCA results showed that the cumulative variance contribution rate of principal components 1-7 was 92.666%. OPLS-DA further identified 13 differential components， which were mainly derived from Polygonati Rhizoma with wine steaming， Rhodiolae Crenulatae Radix Et Rhizoma， prepared Rhei Radix Et Rhizoma and Glycyrrhizae Radix Et Rhizome Praeparata Cum Melle. CONCLUSIONS The established UPLC fingerprint of Jingtian granule is simple， stable and reproducible. Combined with the chemical pattern recognition method， it can effectively reveal the overall quality difference between different batches of Jingtian granule. The quality of Polygonati Rhizoma with wine steaming， Rhodiolae Crenulatae Radix Et Rhizoma， prepared Rhei Radix Et Rhizoma， Dioscoreae Nipponicae Rhizoma， Polyporus， Cinnamomi Ramulus， Glycyrrhizae Radix Et Rhizome Praeparata Cum Melle is the key to the overall quality of Jingtian granule.