Sequencing and Proteomic Analysis of Exosomes from Apheresis Platelets in Different Storage Periods / 中国实验血液学杂志

OBJECTIVE@#To investigate the changes of gene sequencing and proteomics of apheresis platelet (AP) exosomes in different storage periods and predict the function of AP exosomes in different storage periods.@*METHODS@#Platelets at different storage periods of 0 day (D0), 3 day (D3) and 5 day (D5) were collected, exosomes were extracted with Gradient centrifugation; gene sequencing and proteomic analysis were used to analyze the exosomes, and biological functions of platelet exosomes were analyzed and predicted by bioinformatics. Liquid mass spectrometry (LMS) was used to detect the changes and function prediction of exosomes proteins. The small RNA sequencing library was prepared, and the constructed library was sequenced and bioinformatics technology was used for data analysis.@*RESULTS@#AP exosome iTRAQ protein analysis showed that AP exosomes stored in D3 with 55 up-regulated proteins and 94 down-regulated proteins (P<0.05, FC<0.83 or FC>1.2), while AP exosomes stored in D5 with 292 up-regulated proteins and 53 down-regulated proteins (P<0.05, FC<0.83 or FC>1.2) as compared with D0. KEGG pathway analysis showed that the proteins were mainly involved in transport and metabolism, immune system, cancer, membrane transport and other processes. There were statistically significant differences between AP exosome miRNAs in different storage days (P<0.01). The number of miRNA up-regulated and down-regulated was 374 and 255 as compared with the number of platelet exosomes miRNA stored in D3 and D0, while that was 297 and 242 in D5 and D0, and 252 and 327 in D5 and D3, respectively. The target genes of differential platelet exosome miRNAs were analyzed by GO enrichment. Target genes of differential miRNA were mainly involved in membrane composition, mainly played molecular functions binding to proteins, and participated in biological processes of transcriptional regulation.@*CONCLUSION@#The exosome differential proteins and miRNAs in D5 are significantly different from those in the D0 of APs, and they are involved in various biological processes.

Application of RUNX2 gene over expression vector modified exosomes from BMSC combined with calcium carbonate scaffold system in bone defect / 中国骨伤

OBJECTIVE@#To investigate the effect of RUNX2 gene overexpression vector modified exosomes derived from bone marrow mesenchymal stem cells (BMSCs) combined with calcium carbonate scaffold system in bone defect.@*METHODS@#Rabbit BMSCs were used as the research object, and BMSCs were identified by flow cytometry. Construct RUNX2 gene overexpression vector, transfect BMSCs with lentivirus, and collect exosomes by ultracentrifugation. The morphology of exosomes was observed by transmission electron microscope, the expression of exosome marker CD63 was detected by Western blot, and the calcium carbonate scaffold was constructed by three chamber parallel automatic temperature control reaction system. According to whether the RUNX2 gene overexpression vector was transfected or not, the complex of BMSCs and calcium carbonate scaffold was divided into three groups, namely BMSCs group, RUNX2 overexpression group and exosome group. The osteogenic differentiation of BMSCs was detected by oil red O staining and RT-PCR. There were 9 clean adult healthy male New Zealand white rabbits, aged (12.97±1.21) months, with a body weight of (19.3±3.6) kg, with 3 rabbits in each group. The animal model of skull defect was constructed by surgical method, and the repair of bone defect was evaluated by imaging, he staining and Masson staining.@*RESULTS@#The results of flow cytometry showed that the expression of CD29 protein, CD44 protein, CD11b protein and CD45 protein on the surface of BMSCs were 99.5%, 100%, 0.1% and 0.1%, respectively. Transmission electron microscopy showed that the exosomes were bilayer vesicles with a diameter of 50 to 150 nm. Western blot showed that the molecular marker CD63 of exosomes was positive. Oil red O staining showed that the osteogenic differentiation of BMSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group (P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group (P<0.05). MSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group(P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group(P<0.05).@*CONCLUSION@#Compared with RUNX2 gene overexpression vector transfection, extraction of exosomes directly can promote the differentiation of BMSCs into osteoblasts more efficiently, and the combination with calcium carbonate scaffold can better promote the healing of bone defects. So as to provide new ideas and methods for the clinical treatment of bone defects.

Correlation analysis of 3D-FLAIR MRI characteristics of the inner ear and vestibular function in the patients with vestibular neuritis / 中华耳鼻咽喉头颈外科杂志

Objective: The characteristics of 3D-FLAIR MRI images of the inner ear of patients with vestibular neuritis were preliminarily studied to explore the possible pathogenesis of vestibular neuritis, and the correlation analysis was conducted in combination with vestibular function to provide a basis for accurate diagnosis of vestibular neuritis. Methods: A total of 36 patients with vestibular neuritis (VN) from December 2019 to October 2020 were collected from the Vertigo Department of Shandong Provincial ENT Hospital, Cheeloo College of Medicine, Shandong University. There were 36 cases (18 females, 18 males) with unilateral acute vestibular neuritis, 17 cases of left ear and 19 cases of right ear. According to the results of 3D-FLAIR MRI in the inner ears, the patients were divided into the enhanced group and the non-enhanced group (the health side served as the normal control group). The results of vestibular function examination in the two groups were compared. SPSS19.0 software was used for statistical processing to analyze the relationship between the vestibular function and the characteristics of 3D-FLAIR imaging in the inner ears. Results: Abnormal enhancement of 3D-FLAIR was found in 31 cases (86.1%) of the 36 cases, including 14 cases of both vestibular nerve and vestibular terminal organ enhancement, eight cases of superior vestibular nerve enhancement alone, seven cases of vestibular terminal organ enhancement alone, and two cases of cochlear enhancement alone. Observation of abnormal reinforcement of vestibular nerve showed: twenty-one cases of superior vestibular nerve reinforcement, one case of superior and inferior vestibular nerve reinforcement. No abnormalities were found in 3D-FLAIR of inner ear in 5 cases. According to the analysis of vestibular function results, there were 19 cases (52.8%) with total vestibular involvement, sixteen cases (44.4%) with superior vestibular involvement alone, and one case (2.8%) with inferior vestibular involvement alone. Comparison of vestibular function between the five cases (non-enhancement group) and the 31 cases (enhanced group) in the 3D-FLAIR group of the inner ears showed that the CP values of caloric tests in the enhanced group were higher (60.81±3.49 vs 34.12±7.37), with statistically significant difference (t=-2.898, P<0.01). Conclusion: In patients with vestibular neuritis, 3D-FLAIR MRI scan of the inner ear provides visual imaging evidence for clinical practice, considering that the lesion site of vestibular neuritis is not only in the vestibular nerve, but also in the vestibular end organ. Patients with 3D-FLAIR enhanced in the inner ear may have more significant vestibular function damage.

Repair of osteoarthritis in animal model with exosomes derived from BMSCs transfected by the siRNA -Piezo1 through CT navigation / 中国骨伤

OBJECTIVE@#To investigate the effect of the exosomes from bone marrow mesenchymal stem cells (BMSCs) transfected with silence plasmid of Piezol small interference RNA (siRNA)on osteoarthritis (OA) animal model.@*METHODS@#Twenty male SD rats with specific pathogen free (SPF) were selected, ranging in age from 5.46 to 6.96 months, with a mean of (6.21± 0.75) months;and ranging in weight from 385.76 g to 428.66 g, with a mean of (407.21±21.45) g. BMSCs were extracted. The siRNA silencing plasmid of piezo1 was constructed by siRNA technology. After lentivirus was transfected into BMSCs, the exosomes were extracted. At the cellular level, BMSCs were divided into blank plasmid group and siRNA silencing plasmid group according to whether siRNA-Piezo1 was transfected or not. The osteogenic induction ability of siRNA-Piezo1 on BMSCs was detected by RT-PCR and Western blot. At the animal model level, the OA model was established by surgical resection of cruciate ligament of knee joint.According to different treatment schemes, SD rats were divided into 4 groups:blank control group, model group, BMSCs group and exosome group. SD rats in the blank control group were not treated. In the model group, the cruciate ligaments of rats were excised and OA animal model was established. In BMSCs group, BMSCs were injected into knee joint under CT guidance after OA model establishment, and the cell volume was 5×10@*RESULTS@#The lentivirus transfection efficiency was(92.11±4.22)%. RT-PCR showed that the relative expression of Piezo1 mRNA in blank plasmid group was 1.07±0.06, which was significantly different from that of 0.31±0.01 in siRNA silencing plasmid group (@*CONCLUSION@#Piezo1 siRNA silencing vector can promote the differentiation of BMSCs into chondrocytes and effectively inhibit the progression of OA, so as to delay the disease of OA.

Lentivirus mediated siRNA hsa-circ-0000885 transfection of BMSCs and osteoclast co-culture system on cell differentiation, proliferation and apoptosis / 中国骨伤

OBJECTIVE@#To explore the effects of siRNA hsa-circ-0000885 modified bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation, cell proliferation and apoptosis in order to provide new ideas and methods for the clinical treatment of osteoporosis (OP).@*METHODS@#From September 2018 to February 2020, 13 patients with osteoporosis admitted to our hospital were selected as the research objects, including 11 females and 2 males, with an age of (65.45±10.77) years old. After obtaining the informed consent of patients, peripheral blood tissues were extracted. Then the expression level of cir-cRNA in peripheral blood mononuclear cells(PBMC) was detected by circ RNA chip. The expression of circ RNA was silenced by siRNA technology. The BMSCs were transfected with lentivirus. According to the siRNA interference plasmid hsa-circ-0000885, the cells were divided into the blank group, the empty vector group and the siRNA interference group. After 72 hours of treatment, the cell cycle was detected by flow cytometry, the apoptosis level was detected by AV-PI kit, and the osteogenic differentiation ability of BMSCs was detected by ALP staining.@*RESULTS@#The expression of hsa-circ-0000885 in PBMC of patients with osteoporosis was significantly higher than that of healthy controls (@*CONCLUSION@#The lentivirus mediated siRNA hsa-circ-0000885 plasmid transfected into BMSCs and osteoclast co culture system can promote cell proliferation, inhibit apoptosis and promote osteogenic differentiation of BMSCs, which can be used as a potential therapeutic target for OP patients.

Frequency characteristics of horizontal semicircular canals damage and the ultrastructure analysis of crista ampullaris in patients with Meniere's disease / 中华耳鼻咽喉头颈外科杂志

Objective: To investigate the frequency characteristics and the pathological characteristics of the horizontal crista ampullaris in patients with Meniere's disease,and to analyse its structural basis. Methods: Between March, 2019 and November, 2019, seventy-two patients diagnosed as Meniere's disease (27 males and 45 females, aged from 13 to 74 years, with a course of disease ranging from 4 months to 32 years)in Shandong Provincial ENT Hospital were included.Caloric test, sinusoidal harmonic acceleration test (SHA), video-head impulse test (v-HIT), Gadolinium-enhanced inner-ear 3D-FLAIR MRI and pure tone audiometry were conducted in the patients. The function of the horizontal semicircular canal in these patients were analysed as well as its relationship with the degree of endolymphatic hydrops,clinical stage and duration. Light microscopy and transmission electron microscopy were used to observe the ultrastructure of horizontal semicircular canal crista ampullaris from six patients with refractory Meniere's disease who underwent labyrinthectomy. The number of type Ⅰ and type Ⅱ vestibular hair cells, the common pathophysiological changes of horizontal semicircular canal crista ampullaris were investigated in these patients. Statistical analysis was performed using SPSS 19.0. Results: With the increase of detection frequency, the abnormal rate decreased gradually. The abnormal rate of caloric test was 69.4% (50/72), SHA 51.4% (37/72), V-HIT 36.1% (26/72), comparation of the positive rate among the three tests showed statistically significant differences(P<0.05).Neither caloric test nor SHA had correlation with the degree of hydrops(P>0.05), but v-HIT(r=0.434,P<0.01).There was correlation with clinical stage to SHA and v-HIT(r=0.338,0.462,P<0.01), except caloric test(P>0.05).No significant relation was found with caloric test, SHA, v-HIT and course of disease(P>0.05).Morphological observation found abnormal monolayer epithelialization of the horizontal semicircular canal crista ampullaris significantly decreased number of type Ⅱ hair cells compared with type Ⅰhair cells. Hair cells showed perinuclear vacuolization, cytoplasmic vacuoles, mitochondrial electron density increasement and loss of stereocilia. Conclusions: The horizontal semicircular canal damage in the patients with Meniere's disease has a frequency-dependent characteristic, mainly occurres in low frequency area. With progress of the disease, the high frequency area of ampulla will be impaired gradually, and it is related to the degree of endolymphatic hydrops and hearing level. Hair cell injury would be observed,the frequency characteristics may be more associated with the disorder of type Ⅱ hair cells.

Clinical Application of Screening Cell Combination Method in the Prediction of Red Blood Cell Alloantibody / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical application of screening cell combination method in the prediction of red blood cell alloantibody, so as to provide basis for clinical diagnosis.@*METHODS@#From October 2018 to April 2020, 9 680 samples were screened with automatic blood group instrument, 79 patients with positive alloantibodies were identified by 4 sets of screening cells from different manufacturers (referred to as combined method). At the same time, cell panel Panocell-16 was used for comparative analysis. Meanwhile, the combined method was also used to identify the antibodies of 20 samples from National Center for Clinical Laboratories external quality assessment (EQA) in China and 12 samples from WHO EQA.@*RESULTS@#The 79 alloantibodies included anti-Mia antibody (7 cases), anti-M antibody (13 cases), anti-Le@*CONCLUSION@#The combined method can identify the alloantibodies of red blood cells in Chinese population. The screening cells can be used for screening of irregular antibodies without wasting reagents at the same time.

A preliminary study on the mixed teaching of human parasitology based on MOOC resources and the experimental teaching digital platform / 中国血吸虫病防治杂志

The mixed teaching model combines the advantages of traditional teaching and network teaching in the “Internet +” era, which has become one of the important trends in the higher education teaching development. In order to follow this development trend, the human parasitology teaching team makes a reasonable use of modern information techniques, actively promotes the construction and application of online resources, and conducts mixed online and offline teaching based on MOOC resources and the experimental teaching digital platform. This mixed teaching model has shown a positive impact on both teaching and learning among teachers and students; however, students’ personalized independent and deep learning remains unsatisfactory. It is suggested that the online course resources construction, teaching design and digital literacy remain to be increased, so as to create a high-level, innovative and challenging online-offline mixed “golden course”

Comparison of the effect of arthroscopy assisted TightRope plate and Triple-Endobutton plate and Double Endobutton plate in the treatment of acromioclavicular dislocation / 中国骨伤

OBJECTIVE@#To investigate the clinical effect of arthroscopic assisted TightRope plate, Triple-Endobutton plate and Double Endobutton plate in treating of Rockwood type Ⅲ-Ⅴ acromioclavicular dislocation.@*METHODS@#From January 2014 to January 2018, 128 patients with acromioclavicular dislocation were treated by operation. According to the operation plan, the patients were divided into three groups:Double Endobutton group, Triple-Endobutton group and TightRope group. All patients with acromioclavicular dislocation were operated by the same operation team, and the chief surgeon was the same chief physician. General baseline data such as gender, age, operation time, incision length, intraoperative blood loss, VAS score of pain and Constant-Murley shoulder function score were recorded.@*RESULTS@#The wound healed well and no recent complications occurred. One hundred and eleven patients were followed up for 6 to 12(9.1±3.1) months. There was no significant difference on general data among three groups (P >0.05). Among three groups, the operation time of Triple -Endobutton group was the longest, significantly higher than that of other two groups(<0.05);the operation time of TightRope group was the shortest, significantly lower than that of other two groups (<0.05). At 1 month after operation, VAS score comparison of three groups, TightRope group was significantly lower than other two groups, with statistical difference (<0.05). At 12 months after operation of three groups, the Constant-Murley score of TightRope group was significantly higher than that of two group (<0.05). The incidence of incision infectionin TightRope group was significantly lower than that of other two groups(<0.05); the incidence of reduction loss in Double Endobton group was significantly higher than that of other two groups(<0.05), the incidence of reduction loss in TightRope group was significantly higher than that of Triple endobton group(<0.05);the incidence of joint adhesion in TightRope group was significantly lower than that of other two groups(<0.05).@*CONCLUSION@#TightRope plate fixation with arthroscopy is more advantageous than Double Endobutton plate fixation and Triple-Endobutton plate fixation.

Clinical characteristics of patients with indication of cardiac implantable electronic devices implantation complicating with acute pulmonary thromboembolism / 中华心血管病杂志

Objective: To investigate the clinical characteristics of inpatients with the indication of cardiac implantable electronic devices (CIED) therapy and combined acute pulmonary thromboembolism (APTE). Methods: We retrospectively screened 8 641 inpatients who admitted with the indication of CIED implantation in Fuwai Hospital from January 2014 to May 2019. The clinical characteristics, management strategies and clinical outcome were analyzed for patients diagnosed as APTE. Results: APTE were identified in 45 (5‰) patients in this cohort, there were 18(40%) male patients, the average age was (73±8) years old and body mass index was (27±10) kg/m2.Thirty-two (70%) patients were at intermediate-risk and 13 (30%) at low-risk. Anti-coagulation therapy was initiated in 38(84%) patients, and 30 patients underwent CIED implantation (27 pacemaker, 2 CRT and 1 ICD). No postoperative bleeding or pocket hematoma were detected in the 23 patients taking anticoagulation medication before implantation. During an average of (30±7) months' follow up, thrombus was dissolved in 20 patients, hemorrhage complications were observed in 2 patients (1 cerebral hemorrhage and 1 hematuria), anticoagulation therapy was discontinued in these 2 patients. Among 15 patients without immediate CIED implantation and treated with anticoagulation therapy during hospitalization, 2 patients developed complete paroxysmal Ⅲ° atrioventricular block, and recovered after therapy during hospitalization. Seven patients were re-hospitalized for CIED implantation due to bradycardia. Five patients died during follow-up (3 sudden cardiac death, 1 APTE combined with cerebral infarction, and 1 pulmonary infection). Conclusion: APTE is not rare in patients with the indication of CIED implantation, CIED implantation and anti-coagulation therapy are safe for these patients, and transient atrioventricular block could be detected in APTE patients.

A new cadalene sesquiterpene glycoside from Dendrobium nobile / 药学学报

Using column chromatographic and preparative HPLC technologies, we isolated a new sesquiterpene glycoside from the stem of Dendrobium nobile. With spectroscopic techniques including NMR and MS, the new compound was identified as cadalene-12-O-β-D-glucopyranoside. This type of compound was dehydrogenated from cadinane sesquiterpene to achieve a naphthalene ring, and it is rare from a natural resource.

Reaserch Advances on the Role of PI3K/AKT Signaling Pathway and MiRNA in Acute T-Cell Lymphocytic Leukemia--Review / 中国实验血液学杂志

Abstract　　Numerous studies have confirmed that abnormal activation of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway is one of the most common induction mechanisms of T-ALL. In recent years, many literature report that multiple abnormally expressed microRNAs (miRNAs) can participate in the development of T-ALL by regulating the PI3K/AKT signaling pathway. For example, overexpression of miR-19 and miR181a can activate the PI3K/AKT signaling pathway, which leads to the development of T-ALL and induction of chemotherapy drug resistance, as well as the low expression of miR-26b and miR-29a. Apart from the inhibitors and traditional Chinese medicines that target the PI3K/AKT signaling pathway, regulation of the expression of the corresponding miRNA may also be a potential treatment protocol for T-ALL. The mechanisms of PI3K/AKT signaling pathway involved in the development of T-ALL, the role of miRNAs in the PI3K/AKT signaling pathway and the targeted therapy based on this signaling pathway are summarized briefly in this review.

Evaluation and Comparison of Thromboelastography and Conventional Coagulation Tests for Blood Coagulation Function in Children with Henoch-Schönlein Purpura / 中国实验血液学杂志

OBJECTIVE@#To evaluate the coagulation function of children with Henoch-Schönlein purpura (HSP) by thromboelastography (TEG) and conventional coagulation tests (CCTs), and to explore the correlation and consistency of the 2 test methods.@*METHODS@#A total of 468 children with HSP were selected from January 2017 to December 2017 in Beijing Children's Hospital, Capital Medical University. The TEG and CCTs data were analyzed to evaluate coagulation function of children with HSP, meanwhile, the coagulation results were analysed the superiority of the 2 test methods was compared by Pearson correlation and Kappa consistency analysis.@*RESULTS@#There were no clinically significant abnormalities practically in HSP children by TEG and CCTs analysis, except for D-dimer level was elevated (t=9.15, P＜0.001). There were no significant changes for coagulation data from, sex comparison of HSP children (P＞0.05 all), but the coagulation reaction time (R), blood clot formation time (K), α-Angle, CI value, fibrinogen, D-dimer and anti-thrombin III in HSP children with different age groups showed difference (P＜0.05 all), and the blood in children aged 0-2 years old tended to be hypercoagulable. The TEG indexes demonstrated no significant difference in coagulation function of children with HSP each other (P＞0.05). However, CCTs data showed that the blood in children with severe kidney involvement were hypercoagulable. Comparision results of the correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children showed that R was weakly correlated with prothrombin time (PT), International Normalized Ratio (INR) and activated partial thromboplastin time (APTT). There were weak correlation between K, α-Angle and Fib (0.1＜|r|＜0.4 all). There was no obvious consistency between them each other (kappa＜0.4 all).@*CONCLUSION@#The overall changes in coagulation function in children with HSP are not obvious, but the hyperfibrinolysis in hypercoagulable state may exists. Furthermore, younger age and severe kidney involvement may cause hypercoagulation in HSP children. The weakly correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children are furtherly confirmation, and the 2 test methods may be irreplaceable.

Assessment of the Perioperative Risk of Different Operation Times on Renal Angiomyolipoma Patients with Rupture and Hemorrhage / 中山大学学报(医学科学版)

[Objective] To investigate the perioperative risk of different operation times on renal angiomyolipoma (AML) patients with rupture and hemorrhage.[Methods] From January 2006 to December 2016,AML patients with rupture and hemorrhage who receiving surgical treatment were recruited.According to the operation time,patients receiving surgery within 7 days after the hemorrhage were classified as short-term operation group.Patients receiving surgery exceeded 30 days after the hemorrhage were classified as long-term operation group.The general and perioperative data were compared between the two groups.[Results] There were no statistically significant differences in age,tumor size,clinical symptoms and operative methods between the two groups.However,as compared to the long-term operation group,the hemorrhage during surgery [(780±451) vs.(572±913) mL,P=0.029],the volume of transfused blood [(2600 ± 1733) vs.(820±582) mL,P=0.027],the nephrectomy rate (60.0％ vs.22.6％,P=0.027) were significantly increased in the short-term operation group.[Conclusions] A long-term operation group for AML patients with rupture and hemorrhage could have a lower perioperative risk.

Expression characteristics of Piezo1 protein in stress models of human degenerative chondrocytes / 中国骨伤

<p><b>OBJECTIVE</b>To explore the expression characteristics of new mechanosensitive ion channel Piezo1 protein in stress models of human degenerative chondrocytes.</p><p><b>METHODS</b>The stress stimulation model of human degenerative chondrocytes in vitro was constructed. Multi-channel cell stretch stress loading system FX-4000T was used to treat chondrocytes. According to the results of pre-test, the loading frequency of 0.5 Hz and the cell elongation of 20% were loaded. According to cell processing time, it was divided into 0 h, 2 h, 12 h, 24 h and 48 h mechanical stress group. The RT-PCR and Western-blot were used to test the expression of the Piezo1, also the Laser scanning confocal microscope (LSCM) was used to test the intensity of the fluorescence of the Piezo1.</p><p><b>RESULTS</b>(1)The result of the RT-PCR showed that the expression of the Piezo1 in the 2 h group was higher than the 0 h group(13.917, 0.037 1, <0.05). The expression of the piezo1 in the 24 h group was the highest. While the expression of the piezo1 in the 48 h group was lower than the expression of the piezo1 in the 24 h group(13.917, 0.049 5, <0.05). (2)The result of the Western-blot showed that the 2 h group was higher than the 0 h group(19.341, 0.037 1, <0.05). The expression of the 24 h had the highest expression which was higher than the 48 h group(19.341, 0.017 7, <0.05). (3)The Piezo1 protein was extensively expressed in the cytoplasm and nucleus of the nucleus pulposus cells. And with the increase of stress processing time, the fluorescence intensity of the protein also increased.</p><p><b>CONCLUSIONS</b>In human degeneration cartilage cells, the new mechanio sensitive ion channel Piezo1 protein has a trace expression. After loading periodic mechanical tensile force, the expression of Piezo1 protein increases with time dependence.</p>

Clinical analysis of LPFP, PFNA and BPH in treating femoral intertrochanteric fractures in elderly patients / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the clinical results of locking proximal femur plate(LPFP), proximal femoral nail antirotation(PFNA) and bipolar hemiarthroplasty(BPH) in the treatment of femoral intertrochanteric fractures in elderly patients.</p><p><b>METHODS</b>Retrospective analysis of the 86 elderly patients with femoral intertrochanteric fractures during June 2011 and August 2016 were enrolled in this study. The patients were divided into 3 groups: 26 cases in LPFP group included 10 males and 16 females with an average age of (72.54±4.78) years old, 5 cases of type II of Evans, 12 cases of type III and 9 cases of type IV; 39 cases in PFNA group included 17 males and 22 females with an average age of 74.41±5.65, 11 cases of type II, 18 cases of type III and 10 cases of type IV; 21 cases in BPH group included 9 males and 12 females with an average age of 76.23±6.97, 1 case of type II, 10 cases of type III and 10 cases of type IV. The data of three groups were collected for statistical analysis on the following aspects: operation time, intraoperative blood loss, the length of hospital stay, postoperative complications and Harris score of the hip joint function.</p><p><b>RESULTS</b>The wound healed well and no complication occurred. Eighty-four cases were follow-up, while 2 cases were lost, the follow-up rate was 97.6%. The patients were followed up for 22 to 41 months with an average of 26.3 months. For comparison of operation time and time of the hospital stay, BPH group was shorter than the LPFP and PFNA group(=19.782, 21.981,<0.05). For comparison of the intraoperative blood loss, the BPH group was less blood loss than the LPFP and PFNA group(=12.976,<0.05), while there was no significant means between the LPFP and PFNA group(=10.879,>0.05). For comparison of the postoperative complications, LPFP group was higher than BHP and PFNA group(=30.976,<0.05), while there was no significant means between LPFP and PFNA group(=9.902,>0.05). For comparison of the Harris score of the hip joint at 1 year after operation, BPH and PFNA group were both higher than LPFP group(=19.692,<0.05), while there was no significant means between BPH and PFNA group (=4.971,>0.05).</p><p><b>CONCLUSIONS</b>For elder patient with femoral intertrochanteric fracture, BPH could be the best optional for the shorter operation time and less complications. And the PFNA also could be optional choice for the patients while the LPFP was forbidden for the elder patients.</p>

Influence of drug metabolizing enzyme inhibitors on liver injury susceptibility to trans-2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside / 药学学报

By using the drug metabolizing enzyme inhibitors, the effects of metabolic factors on potential liver injury induced by the main component, trans-2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(trans-SG), in Polygonum multiflorum was investigated. The main metabolic enzyme isoforms involved in trans-SG metabolism were also screened. The results showed that trans-SG at the dosage 31 mg·kg-1 did not cause liver injury; and the combination of trans-SG with the phase I metabolic enzyme inhibitor, 1-benzylimidazole (10 mg·kg-1), did not change the degree of liver injury(compared with LPS + trans-SG group, P > 0.05). However, the combination of trans-SG with phase II metabolic enzyme inhibitor, ketoconazole(35 mg·kg-1), significantly increased the degree of liver injury(compared with LPS + trans-SG group, P < 0.05). The phase I metabolites of trans-SG were not detected in human liver microsomes phase I metabolism system, while the phase II trans-SG metabolites were detected in recombinant human UGT isozymes phase II metabolism system. Six isoforms of uridine diphosphate glucuronate transferase(UGT)exhibited abilities to metabolize trans-SG and the order of metabolic ability was: UGT1A1 > UGT1A9 > UGT1A7 > UGT1A10 > UGT2B7 > UGT1A8. The results showed that trans-SG was mainly metabolized by UGT in phase II metabolism. The inhibition of drug metabolizing enzymes of phase II can increase the liver injury susceptibility of trans-SG, which provides a reference to the evaluation of susceptible factors and drug incompatibility research of Polygonum multiflorum.

Regulatory Effect of Qushi Huayu Recipe on Gene Expression Profiles of Fatty Liver Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the intervention and mechanism of Qushi Huayu Recipe (QHR) on gene expression profiles in high lipid diet induced fatty liver rats.</p><p><b>METHODS</b>Fatty liver model was prepared in 20 male SD rats using single high fat diet (88% common forage +2% cholesterol +10% lard). Four weeks after modeling they were divided into the model group and the QHR group according to random digit table, 10 in each group. QHR (at 0. 93 g crude drug/100 g body weight) and distilled water was respectively to rats in the QHR group and the model group by gastrogavage while modeling, once per day. Meanwhile, 10 SD male rats were recruited in a normal group, administered with equal volume of distilled water by gastrogavage. At the end of week 8 all rats were sacrificed, and blood and livers were collected for subsequent analysis. Contents of liver triglyceride (TG) and free fatty acid (FFA) , activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using biochemical assay. Pathological changes of liver tissue were observed using H&E and oil red O stain. Liver gene expressions were detected by Affymetrix gene expression profiles. Differentially expressed genes were compared between the QHR group and the model group, functions of differentially expressed genes and signal pathways involved analyzed. Ten differentially expressed genes involved in glycolipid metabolism with fold change more than 2 were selected for verification by real-time PCR.</p><p><b>RESULTS</b>(1) Compared with the normal group, contents of liver TG and FFA, and serum activities of ALT and AST obviously increased in the model group (P <0. 01). Compared with the model group, contents of liver TG and FFA, and activities of ALT and AST obviously decreased in the QHR group (P <0. 05, P <0. 01). QHR could reduce high fat induced fatty degeneration of liver cells , alleviate inflammation, and improve pathological changes of liver tissue. (2) Compared with the model group, there were 80 differentially expressed genes (with fold change > 2, P < 0.05) with clear functions and appointed gene names, including 44 up-regulated and 36 down-regulated genes. Eighty genes were involved in 27 signal pathways with statistical difference, including glycerolipid metabolism, adipocytokine signaling pathway, insulin signal pathway, drug metabolism signal pathway, etc (P < 0.05). (3) RT-PCR results of 10 glycolipids metabolism regulating genes such as Gk, Scd1, Gpat2, G6pc, Irs1, and so on showed that all RT-PCR genes were completely coincide with up-regulated or down-regulated tendency in results of gene chips. 80% genes had approximate fold change.</p><p><b>CONCLUSION</b>QHR could regulate gene expressions related to fat metabolism, carbohydrate metabolism, anti-lipid peroxidation, and drug metabolism in high fat diet induced fatty liver rats, and its comprehensive pharmacological actions could be manifested.</p>

Influence of metal ions on stability of 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside contained in Polygoni Multiflori Radix / 药学学报

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG. The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration-dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2-O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.

Influence of metal ions on stability of 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside contained in Polygoni Multiflori Radix / 药学学报

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG . The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration- dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe3+ and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2- O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.