1.Polymerase chain reaction-based assays facilitate the breeding and study of mouse models of Klinefelter syndrome.
Hai-Xia ZHANG ; Yu-Lin ZHOU ; Wen-Yan XU ; Xiao-Lu CHEN ; Jia-Yang JIANG ; Xiao-Man ZHOU ; Zeng-Ge WANG ; Rong-Qin KE ; Qi-Wei GUO
Asian Journal of Andrology 2022;24(1):102-108
Klinefelter syndrome (KS) is one of the most frequent genetic abnormalities and the leading genetic cause of nonobstructive azoospermia. The breeding and study of KS mouse models are essential to advancing our knowledge of the underlying pathological mechanism. Karyotyping and fluorescence in situ hybridization are reliable methods for identifying chromosomal contents. However, technical issues associated with these methods can decrease the efficiency of breeding KS mouse models and limit studies that require rapid identification of target mice. To overcome these limitations, we developed three polymerase chain reaction-based assays to measure specific genetic information, including presence or absence of the sex determining region of chromosome Y (Sry), copy number of amelogenin, X-linked (Amelx), and inactive X specific transcripts (Xist) levels. Through a combined analysis of the assay results, we can infer the karyotype of target mice. We confirmed the utility of our assays with the successful generation of KS mouse models. Our assays are rapid, inexpensive, high capacity, easy to perform, and only require small sample amounts. Therefore, they facilitate the breeding and study of KS mouse models and help advance our knowledge of the pathological mechanism underlying KS.
Animals
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Azoospermia
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In Situ Hybridization, Fluorescence
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Karyotyping
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Klinefelter Syndrome/genetics*
;
Mice
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Polymerase Chain Reaction
2.Mechanism of Xinfeng Capsules improving rheumatoid arthritis based on CD19~+B cells regulating FAK/CAPN/PI3K pathway.
Lei WAN ; Jian LIU ; Chuan-Bing HUANG ; Lei ZHAO ; Xi CHEN ; Hai-Xia FAN ; Yao GE ; Tian-Yang LIU ; Lei LIU ; Zheng LI ; Zi-Heng ZHU ; Xi-Meng MA ; Shu LI
China Journal of Chinese Materia Medica 2021;46(14):3705-3711
To observe the effect of Xinfeng Capsules on rheumatoid arthritis (RA) B lymphocytes,inflammatory mediators,FAK/CAPN/PI3K pathway,in order to explore the mechanism of Xinfeng Capsules in improving clinical symptoms of RA.Joint and systemic symptoms of RA patients were observed,and laboratory indicators[hemoglobin (HGB),platelet count (PLT),erythrocyte sedimentation (ESR),immunoglobulin (Ig) G,Ig A,Ig M,rheumatoid factor (RF),anti-cyclic citrulline antibody (CCP-AB),C-reactive protein (CRP)]were detected.ELISA was used to detect serum interleukin (IL)-1β,IL-10,IL-33,chemokine 5 (CCL5),and vascular endothelial growth factor (VEGF).CD3~-CD19~+B cells were measured by flow cytometry.Western blot was used to detect FAK,p-FAK,CAPN,PI3K protein.The results showed that Xinfeng Capsules could significantly alleviate RA joint and systemic symptoms and improve clinical efficacy.And Xinfeng Capsules could increase HGB,decrease PLT,CCP-AB,CRP,ESR index,upregulate IL-10 expression,and down-regulate IL-1β,IL-33,CCL5,VEGF,CD3~-CD19~+B cells,FAK,p-FAK,CAPN,PI3K expressions (P<0.01).Based on the above results,Xinfeng Capsules may reduce the expression of CD3~-CD19~+,regulate the balance of inflammatory cytokines and chemokines,inhibit abnormal activation of FAK/CAPN/PI3K pathway,and improve clinical symptoms of RA.
Arthritis, Rheumatoid/drug therapy*
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B-Lymphocytes
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Capsules
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Drugs, Chinese Herbal
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Humans
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Phosphatidylinositol 3-Kinases
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Vascular Endothelial Growth Factor A
3.Analysis of peptides and proteins from Asini Corii Colla using nano LC-Q-Exactive-MS/MS.
Xia-Yan CHU ; Un Meng-Ru S ; Gui-Hua JIA ; Yan-Tao WANG ; Jian-Jun GU ; Hai-Bin LIU ; Guang-Bo GE
China Journal of Chinese Materia Medica 2021;46(24):6422-6434
This paper aims to systematically analyze the peptides and proteins from Asini Corii Colla(ACC) through shotgun proteomics. After high-pH reversed-phase fractionation, the proteins and peptides in the hydrolysate of ACC were further separated by nano LC-Q-Exactive-MS/MS under the following conditions: Thermo Scientific EASY column(100 μm×2 cm, 5 μm, C_(18)) as precolumn, Thermo Scientific EASY column(75 μm×100 mm, 3 μm, C_(18)) for solid phase extraction, gradient elution with 0.1% formic acid in water(mobile phase A) and 84% acetonitrile in water containing 0.1% formic acid(mobile phase B), and MS in positive ion mode. Based on Uniprot_Equus caballus, MS data, and literature, 2 291 peptides were identified from ACC by MaxQuant, with 255 Maillard reactions(AML, CML, CEL)-modified peptides identified for the first time. Through alignment, the peptides were found to belong to 678 equine proteins. In conclusion, the combination of nano LC-Q-Exactive-MS/MS and shotgun proteomics achieved rapid and accurate identification of the proteins and peptides in ACC, which provides the key information and new insights for further investigation of chemicals and effective substances in ACC.
Animals
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Chromatography, Liquid
;
Horses
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Peptides
;
Proteins
;
Proteomics
;
Tandem Mass Spectrometry
4.Renal artery involvement: independent risk factors of KDIGO stage 3 in acute renal injury after moderate hypothermic circulatory arrest in acute Stanford type A aortic dissection
Yipeng GE ; Chengnan LI ; Yonglang ZHONG ; Yu XIA ; Fucheng XIAO ; Ou Hai' HU ; Tie ZHENG ; Junming ZHU ; Lizhong SUN
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(6):335-338
Objective:To explore whether renal artery involvement is an independent risk factor of acute renal injury (AKI) KDIGO stage 3 after moderate hypothermic circulatory arrest in patients with acute Stanford type A aortic dissection.Methods:From December 2015 to October 2017, 492 consecutive patients with acute Stanford A-type aortic dissection received surgical treatment, 486 of them were included in the study. All patients underwent aortic CTA to determine the extent of aortic dissection and renal artery involvement. According to the standard of Improving Global Outcomes (KDIGO), the renal function of patients after operation was graded. The risk factors of AKI KDIGO stage 3 were analyzed.Renal artery involvement and other risk factors were included in univariate analysis, and significant variables in univariate analysis were included in multivariate logistic regression analysis.Results:In 492 patients, 40 (8.13%) died in hospital, of which 6 died of severe bleeding during operation or failed to wean from cardiopulmonary bypass which lead to unable to leave the Weaning from cardiopulmonary bypass and these 6 patients were excluded in the research. Among 486 patients included in the study, 251 (51.64%) had AKI. Among them, 83 (17.08%) were in the KDIGO stage 1, 56 (11.52%) in stage 2 and 112 (23.05%) in stage 3.The results of univariate analysis showed that there were significant differences in renal artery involvement, age, time from onset to operation, D-dimer, leukocytes and platelets in peripheral blood, creatinine clearance rate, time of cardiopulmonary bypass during operation and aortic cross-clamping time( P>0.05). The above risk factors were included in multivariate logistic regression. The results showed that preoperative renal artery involvement ( OR=1.94, P=0.02), age ( OR=1.03, P=0.02), creatinine clearance rate<85 ml/min ( OR=2.28, P=0.001), and intraoperative cardiopulmonary bypass time ( OR=1.01, P=0.02) were independent risk factors. The incidence of AKI in patients with renal artery involvement was 54.65%, significantly higher than 41.98% in patients without renal artery involvement ( P>0.05). Conclusion:Renal artery involvement is an independent risk factor of AKI KDIGO stage 3 after moderate deep hypothermic circulatory arrest of acute Stanford type A aortic dissection.
5.Protection of Addition and Subtraction Therapy of Ditantang Combined with Taohong Siwutang to Brain Nerve of Patients with Cerebral Infarction and (Syndrome of Phlegm and Blood Stasis Blocking Collaterals) During Early Recovery
Yong-xia CHANG ; Jiao LI ; Wen-li HOU ; Lei GE ; Yin WANG ; Teng WANG ; Qiu-yun MA ; Hai-chao MENG ; Xiao-hong ZUO ; Wan-seng CUI
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(1):135-140
Objective:To discuss clinical effect of addition and subtraction therapy of Ditantang combined with Taohong Siwutang to cerebral infarction and syndrome of phlegm and blood stasis blocking collaterals during early recovery, and to study protection to brain nerve. Method:One hundred and fifty-two patients were randomly divided into control group (76 cases) and observation group (76 cases) by random number table, 71 patients in control group completed the therapy (5 patients were falling off, missing visit or eliminated), and 70 patients in observation group completed the therapy. Both groups' patients got comprehensive rehabilitation measures. Patients in control group got Zhongfeng Huichun pills, 1.5 g/time, 3 times/day. Patients in observation group got addition and subtraction therapy of Ditantang combined with Taohong Siwutang in the morning and at night, 1 dose/day. The treatment was continued for 12 weeks. Before and after treatment, scores of degree of neurological deficit, Barthel (BI) index, Fugl-Meyer scale (FMA), modified Rankin scale (MRS) and syndrome of phlegm and blood stasis blocking collaterals were graded. And levels of malondialdehyde (MDA), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), advanced oxidation protein products (AOPP), vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF) and neuron specific enolase (NSE). And cerebral hemodynamics were detected, and peak flow velocity (VS), vascular resistance index (RI), pulsatility index (PI) and cerebrovascular reserve function (CVR) were recorded. Safety was evaluated. Result:After the 6th week and 12th week of treatment, scores of degree of neurological deficit, BI, FMA, MRS, syndrome of phlegm and blood stasis blocking collaterals, AOPP, MDA, NSE, RI and PI were lower than those in control group (
6.Mycoplasma pneumoniae Macrolide Resistance and MLVA Typing in Children in Beijing, China, in 2016: Is It Relevant?
Wei Hai DOU ; Jun Xiu TIAN ; Li De XIN ; Ran WEI ; Wei ZHOU ; Hong WANG ; Guang Xuan QIN ; Yan Jun SHAO ; Ping Bao XU ; Xia Li GE ; Wei Da SHI
Biomedical and Environmental Sciences 2020;33(12):916-924
Objective The aim of this study is to investigate the macrolide resistance rate and molecular type withmultiple-locus variable-number tandem-repeat analysis (MLVA) of Mycoplasma pneumoniae of Beijing in 2016 in pediatric patients. Methods Real-time quantitative polymerase chain reaction (PCR) was used to identify M. pneumoniae, and MLVA was performed. The domain V of the 23S rRNA was sequenced to detect macrolide-resistant point mutations. We also investigated the activities of antibiotics against M. pneumoniae isolates in vitro. Results The PCR detection rate of M. pneumoniae in children in Beijing was 40%, and the macrolide resistance rate was 66%. The A2063G mutation in the 23S rRNA V region is the dominant mutation (137/146, 93.84%), whereas the A2064G mutation is rare (9/146, 6.16%). Seventy-three samples were typed successfully by MLVA typing, including 86.3% (63/73) were MLVA type 4-5-7-2, and 13.7% (10/73) were MLVA type 3-5-6-2. No other types were found. No strains were resistant to levofloxacin or tetracycline. Conclusion In 2016, a specific decrease in the macrolide resistance rate occurred in Beijing. The detection rate and macrolide resistance rate of outpatients are lower than those of inpatients. The A2063G mutants M. pneumoniae have high levels of resistance to erythromycin and azithromycin. The primary MLVA type is 4-5-7-2, followed by 3-5-6-2. No other MLVA types were detected. No strains resistant to tetracycline or levofloxacin were found in vitro.
7.Expression Levels of HES1, C-MYC and NF-kB in Peripheral Blood of Patients with T Cell Acute Lymphoblastic Leukemia and Their Significance.
Hai-Li GAO ; Wei LIU ; Liang TIAN ; Yan-Ge LI ; Hui-Xia LI ; Yan-Na MAO
Journal of Experimental Hematology 2019;27(5):1449-1454
OBJECTIVE:
To analyze and investigate the expression levels of HES1, C-MYC and NF-kB in peripheral blood of patients with T cell acute lymphoblastic leukemia (T-ALL) and their significance.
METHODS:
Sixty patients with T-ALL and 60 patients with acute myelogenous leukemia (AML) diagnosed in our hospital from June 2012 to March 2015 were enrolled in T-ALL group and AML group, respectively. Another 30 healthy people were enrolled in the control group. Peripheral blood was collected to detect the expression levels of HES1, C-MYC and NF-kB by RT-PCR. The general data and the expression of HES1, C-MYC and NF-kB in peripheral blood were compared among the patients with different type of leukemia, cytogenetical types and different prognosis.
RESULTS:
There was no significant difference in baseline data, such as age and sex among the 3 groups (P>0.05). The Hb level, WBC and Plt count, BM blast cell ratio in T-ALL and AML groups all were significantly higher than those in control group (P<0.01), but there were no statistical difference in above-mentioned indicators between T-ALL and AML groups (P>0.05). The expression levels of HES1, C-MYC and NF-kB in peripheral blood among 3 groups were significantly differenct (P<0.01), the expressions levels of HES1, C-MYC and NF-kB in T-ALL and AML groups were significantly higher than those in control were significantly group (P<0.01), moreover, the expression levels of above-mentional indicators in T-ALL groups were significantly higher than than those in AML group (P<0.01). The expression levels of HES1, C-MYC and NF-kB iin T-ALL patients with poor prognosis were significantly higher than those in T-ALL patients with favorable prognosis (P<0.01); the expression levels of HES1, C-MYC and NF-kB in peripheral blood of patients with different theraptic efficacy were follow: complete remission group<partial remission group<no remission group (P<0.01).
CONCLUSION
The HES1, C-MYC and NF-kB are highly expressed in peripheral blood of the patients with T-ALL, moreover, the expression levels maybe different, because of the cytogenetic, and theraptic efficacy.
Humans
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Leukemia, Myeloid, Acute
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NF-kappa B
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Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
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Remission Induction
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T-Lymphocytes
;
Transcription Factor HES-1
8.Influence of implants prepared by selective laser melting on early bone healing.
Jing Yin LIU ; Fei CHEN ; Yan Jun GE ; Ling WEI ; Shao Xia PAN ; Hai Lan FENG
Journal of Peking University(Health Sciences) 2018;50(1):117-122
OBJECTIVE:
To evaluate the influence of the rough surface of dental implants prepared by selective laser melting (SLM) on early bone healing around titanium implants.
METHODS:
A total of sixteen titanium implants were involved in our research, of which eight implants were prepared by SLM (TIXOS Cylindrical, Leader-Novaxa, Milan, Italy; 3.3 mm×10 mm, internal hex) and the other eight were sandblasted, large-grit and acid-etched (SLA) implants (IMPLUS Cylindrical, Leader-Novaxa, Milan, Italy; 3.3 mm×10 mm, internal hex). All of the dental implants were inserted into the healed extraction sockets of the mandible of two adult male Beagle dogs. Half of the dental implants were designed to be healed beneath the mucosa and the other half were intended to be healed transgingivally and were immediately loaded by acrylic resin bridge restoration. Three types of tetracycline fluorescent labels, namely calcein blue, alizarin complexone and calcein, were administered into the veins of the Beagle dogs 2, 4, and 8 weeks after implant placement respectively for fluorescent evaluation of newly formed bone peri-implant. Both Beagle dogs were euthanized 12 weeks after implant insertion and the mandible block specimens containing the titanium implants and surrounding bone and soft tissue of each dog were carefully sectioned and dissected. A total of 16 hard tissue slices were obtained and stained with toluidine blue for microscopic examination and histomorphometric measurements. Histological observation was made for each slice under light microscope and laser scanning confocal microscope (LSCM). Comparison on new bone formation around titanium implants of each group was made and mineral apposition rate (MAR) was calculated for each group.
RESULTS:
Dental implants prepared by selective laser melting had achieved satisfying osseointegration to surrounding bone tissue after the healing period of 12 weeks. Newly formed bone tissue was observed creeping on the highly porous surface of the SLM implant and growing into the pores of surface structure. Higher MAR values were shown for SLM implants compared with SLA implants (P<0.01).
CONCLUSION
Dental implants prepared by selective laser melting could promote early bone healing and improve mineral apposition rate.
Animals
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Dental Implantation, Endosseous
;
Dental Implants
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Dental Prosthesis Design
;
Dogs
;
Male
;
Mandible
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Osseointegration
;
Surface Properties
;
Titanium
9.Evaluation of irrational drug use in elderly inpatients based on the list of potentially inappropriate medication for the Chinese aged people
Jun CAI ; Li NIE ; Hai-Xia ZHANG ; Jin-Ping ZHANG ; Wei-Hong GE
The Chinese Journal of Clinical Pharmacology 2018;34(3):375-377
Objective To provide the reference for rational use of drugs in elderly inpatients by retrospective study of the potentially inappropriate medication (PIM)of elderly inpatients.Methods Two hundred and fourteen patients aged over 65 years old were collected,potentially inappropriate medicine was defined by the list of potentially inappropriate medication for the Chinese aged people.Results 77.10% (165 cases/ 214 cases) patients had potentially inappropriate medication and the number of PIM was 269.The PIM was mainly related to clopidogrel (32.71%),insulin (29.74%),quetiapine (7.81%),and zolpidem (5.95%).Conclusion The incidence of potentially inappropriate medication in elderly inpatients was high in the hospital.Some measures should be taken to reduce the incidence of PIM and protect the elderly inpatients from irrational drug use.
10.Xinfeng capsule for the treatment of rheumatoid arthritis patients with decreased pulmonary function--a randomized controlled clinical trial.
Lei WAN ; Jian LIU ; Chuan-bing HUANG ; Yuan WANG ; Xi CHEN ; Wan-dong ZHANG ; Gui-zhen WANG ; Hai-xia FAN ; Yao GE ; Rui-lian CHEN ; Yun-xiang CAO ; Rui-kai ZONG ; Tian-yang LIU
Chinese journal of integrative medicine 2016;22(3):168-176
OBJECTIVETo determine the effectiveness and safety of Xinfeng Capsules (XFC) for the treatment of rheumatoid arthritis (RA) patients with decreased pulmonary function.
METHODSThis was a randomized controlled clinical trial of 80 RA patients. Participants were assigned to the trial group (40 cases) and the control group (40 cases) by block randomization. The trial group was treated with XFC, three pills each time three times daily for 2 months. The control group was treated with tripterygium glycoside (TPT), two pills each time three times daily for 2 months. Both groups were followed up after 2 months. The clinical effects, changes in joint and pulmonary function, and quality of life before and after treatment were observed; safety indices were also evaluated.
RESULTSPain, swelling, tenderness, and duration of morning stiffness of joints were obviously decreased after treatment in both the trial and the control groups compared with baseline (P<0.01). Compared with before treatment, hand grip strength increased significantly after treatment in the trial group (P=0.0000); pulmonary function parameters such as forced expiratory volume in the first second of expiration/forced vital capacity (FEV1/FVC), 50% of the expiratory flow of forced vital capacity (FEF50), carbon monoxide diffusing capacity (DLco) were increased (P<0.01 or P<0.05); measures of quality of life such as role-physical, body pain, vitality and mental health were also improved after treatment in the trial group (all P<0.05). Joint swelling in the trial group decreased compared with the control group (P=0.0043), while hand grip strength was increased after treatment (P=0.0000). The increase in FEF50, DLco, and the dimensions of quality of life such as vitality and mental health were all significantly greater in the trial group than the control group (P<0.05 or P<0.01).
CONCLUSIONSXFC not only relieved joint pain in RA patients, but also significantly improved the ventilation and diffusion function of the lungs. Therefore, XFC could improve the whole body function and enhance the quality of life of RA patients.
Adult ; Aged ; Arthritis, Rheumatoid ; blood ; drug therapy ; pathology ; physiopathology ; Blood Sedimentation ; C-Reactive Protein ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Joints ; pathology ; Male ; Middle Aged ; Quality of Life ; Respiratory Function Tests ; Surveys and Questionnaires ; Treatment Outcome

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