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1.
Asian Journal of Andrology ; (6): 73-77, 2022.
Article in English | WPRIM | ID: wpr-928539

ABSTRACT

This study analyzed the effects of male age and abstinence time on semen quality and explored the best abstinence time for Chinese males among different age groups. Semen parameters, including sperm kinetics, morphology, and DNA fragmentation index (DFI), were reviewed from 2952 men. Samples were divided into six age groups (≤25 years, 26-30 years, 31-35 years, 36-40 years, 41-45 years, and >45 years) and were divided into six groups according to different abstinence time (2 days, 3 days, 4 days, 5 days, 6 days, and 7 days). The differences in semen quality between the groups were compared, and the effect of age and abstinence time on semen quality was analyzed. Significant differences were observed in semen volume, progressive motility (PR), and DFI among the age groups (all P < 0.05), and no significant differences were observed in sperm morphological parameters (all P > 0.05). There were significant differences in semen volume, PR, and DFI among different abstinence time groups (all P < 0.05) and no significant differences in sperm morphological parameters (all P > 0.05). Pearson analysis showed that male age and abstinence time were both significantly correlated with sperm kinetics and DFI (both P < 0.05), while no significant correlation was found with sperm morphological parameters (all P > 0.05). The box plots and histograms of men's age, abstinence time, and semen quality show that most semen quality parameters differ significantly between the 2 days and 7 days abstinence groups and other groups at different ages. Except for the sperm morphology parameters, sperm kinetic parameters and sperm DFI are linearly related to male age and abstinence time.


Subject(s)
Adult , DNA Fragmentation , Humans , Male , Retrospective Studies , Semen , Semen Analysis , Sperm Count , Sperm Motility , Spermatozoa
2.
Article in Chinese | WPRIM | ID: wpr-927982

ABSTRACT

Based on ITS sequences, the molecular identification of Cordyceps cicadae and Tolypocladium dujiaolongae was carried out, and high-performance liquid chromatography(HPLC) fingerprint combined with chemical pattern recognition method was established to differentiate C. cicadae from its adulterant T. dujiaolongae. The genomic DNA from 10 batches of C. cicadae and five batches of T. dujiaolongae was extracted, and ITS sequences were amplified by PCR and sequenced. The stable differential sites of these two species were compared and the phylogenetic tree was constructed via MEGA 7.0. HPLC was used to establish the fingerprints of C. cicadae and T. dujiaolongae, and similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminant analysis(PLS-DA) were applied to investigate the chemical pattern recognition. The result showed that the sources of these two species were different, and there were 115 stable differential sites in ITS sequences of C. cicadae and T. dujiao-longae. The phylogenetic tree could distinguish them effectively. HPLC fingerprints of 18 batches of C. cicadae and 5 batches of T. dujiaolongae were established. The results of CA, PCA, and PLS-DA were consistent, which could distinguish them well, indicating that there were great differences in chemical components between C. cicadae and T. dujiaolongae. The results of PLS-DA showed that six components such as uridine, guanosine, adenosine, and N~6-(2-hydroxyethyl) adenosine were the main differential markers of the two species. ITS sequences and HPLC fingerprint combined with the chemical pattern recognition method can serve as the identification and differentiation methods for C. cicadae and T. dujiaolongae.


Subject(s)
Chromatography, High Pressure Liquid/methods , Cordyceps/genetics , Hypocreales , Phylogeny
3.
China Pharmacy ; (12): 1183-1188, 2022.
Article in Chinese | WPRIM | ID: wpr-924070

ABSTRACT

OBJECTIVE To study the effects of Shenfu yixin granule on mitochondrial autophagy of cardiomyocytes in rats with heart failure after acute myocardial infarction. METHODS The model of heart failure after acute myocardial infarction was established by ligaturing the anterior descending branch of the left coronary artery in rats. The model rats were divided into model group,Shenfu yixin granule low-dose and high-dose groups (1.76,8.8 g/kg),Fosinopril sodium tablets group (positive control ,4 mg/kg),sham operation group was set up (only threading without ligation at the same position ),with 8 rats in each group. After 4 weeks of drug intervention ,the hemodynamic indexes of rats in each group were measured by physiological recorder. The pathological changes of myocardial tissue were observed in each group. The level of oxidative stress in cardiomyocytes , mitochondrial membrane potential ,protein expression of PTEN-induced putative kinase 1(PINK1),E3 ubiquitin ligase Parkin and ubiquitin binding protein P 62 in myocardial tissue of rats in each group were detected. RESULTS Compared with sham operation group ,the pathological injuries such as myocardial fiber morphology disorder and inflammatory cell infiltration were serious. The left ventricular end systolic pressure (LVESP),maximum rate of rise of left ventricular internal pressure (+dp/dtmax), maximun rate of decrease of left ventricular internal pressure (-dp/dtmax),total antioxidant capacity ,mitochondrial membrane potential,PINK1,Parkin and P 62 protein expression were significantly decreased in model group (P<0.01). The left ventricular end diastolic pressure (LVEDP),the level of reactive oxygen species and the activity of reduced nicotinamide adenine dinucleotide phosphate in left ventricular ischemic cardiomyocytes were significantly increased (P<0.01). Compared with model group ,the pathological injuries of myocardial tissue in intervention groups were alleviated ,and above indexes were improved in varying degrees(P<0.01 or P<0.05). CONCLUSIONS Shenfu y ixin granule can reduce the level of oxidative stress and alleviate heart failure after acute myocardial infarction ,which may be related to the activation of Parkin-dependent pathway to strengthen mitochondrial autophagy and reduce mitochondrial dysfunction.

4.
Article in Chinese | WPRIM | ID: wpr-906311

ABSTRACT

Objective:To explore the macroscopic medication rule of Chinese medicine for the treatment of primary liver cancer and provide references for clinical medication. Method:The databases of CNKI,VIP, and Wanfang Data were searched for research articles published from September 1959 to June 2019 with the terms of "Chinese medicine" and "liver cancer". A database was established based on the collected Chinese medicinal prescriptions for the treatment of primary liver cancer. The frequency,clustering, and association rules were analyzed by Excel, etc. Result:In this study,106 effective articles were included,and after the modified prescriptions were removed, 92 effective prescriptions were screened out,involving 281 Chinese herbal medicines used for 1 181 times in total. The top 5 high-frequency drugs were Poria (deficiency-tonifying),Astragali Radix (heat-clearing),Bupleuri Radix (blood-activating and stasis-resolving),Paeoniae Radix Alba (urination-promoting and dampness-draining), and Codonopsis Radix (Qi-regulating). The analysis of drug flavor with a frequency higher than 10 showed that most of the drugs were sweet,bitter, and pungent in flavor,cold,warm, and plain in nature,and acted on spleen and liver meridians. Four combinations and 10 herbal pairs were obtained by the cluster analysis of high-frequency drugs and association analysis, respectively. The high-frequency drugs and potential herbal pairs were classified targeting the specific clinical syndromes in different stages of liver cancer. Conclusion:Replenishing Qi, invigorating spleen,clearing heat, removing toxin,activating blood, and resolving stasis were the basic principles for the treatment of primary liver cancer. The combination of those drugs was the main therapeutic strategy. In addition,the resulting 10 potential herbal pairs from high-frequency drugs and cluster analysis could inspire the clinical treatment of primary liver cancer in different clinical stages with various clinical syndromes, which was of reference value for the clinical medication.

5.
Article in Chinese | WPRIM | ID: wpr-882752

ABSTRACT

Objective:To summarize the experience in diagnosis and treatment of primary cardiac tumors in pediatric patients.Methods:Retrospectively analyzing 7 pediatric patients who were suspected as primary cardiac tumors and diagnosed and treated in Department of Heart Center, Children′s Hospital of Dalian Medical University from August 2013 to February 2019.All patients underwent echocardiography and other examinations, so as to confirm the diagnosis and the treatment plan was chosen based on the size and location of the tumor.All patients were followed up after discharge.Results:A total of 7 patients were diagnosed as primary cardiac tumors by echocardiography, among which 5 cases underwent surgical treatment, and 2 cases were diagnosed with tuberous sclerosis without surgery.In children undergoing surgery, 1 patient underwent autologous heart transplantation to remove the tumor, 1 patient had arrhythmia, 1 patient had mitral regurgitation after surgery, and the mitral regurgitation was corrected again.The remaining children had no adverse complications and were discharged successfully.Histologic examination revealed rhabdomyoma in 4 patients, and fibroma in 1 patient.The patients were followed up for 2-66 months after discharge, and no tumor recurrence was observed in the children who performed surgery.There was a trend of spontaneously regress of cardiac tumor in 2 patients without surgery.Conclusions:Echocardiography is the first choice for the diagnosis of primary cardiac tumors in Pediatric patients.Rhabdomyoma is the most prevalent histologic type of primary cardiac tumors, and tuberous sclerosis should be excluded during the diagnosis process.Patients with tuberous sclerosis selected conservative treatment, and surgical treatment was selected for children with obvious symptoms.According to the location and size of lesion, therapy strategies should be chosen and autologous heart transplantation can be adopted to remove the tumor for children with large tumors.Autologous heart transplantation to remove the tumor is a good surgical treatment.

6.
Article in Chinese | WPRIM | ID: wpr-873333

ABSTRACT

Objective::To identify and analyze the chemical constituents in Bufei Jianpi formula by UPLC-Q-TOF-MS/MS. Method::An Agilent Poroshell SB-C18 column (4.6 mm×100 mm, 2.7 μm) was used with a mobile phase system of 0.1% formic acid solution (A)-acetonitrile (B) for gradient elution (0-10 min, 3%B; 10-100 min, 3%-50%B; 100-120 min, 50%-100%B) under positive ion mode and water (A)-acetonitrile (B) for gradient elution (0-5 min, 3%B; 5-60 min, 3%-100%B) under negative ion mode, the flow rate was 0.6 mL·min-1, and the column temperature was 30 ℃. Mass spectrometric data were obtained under electrospray inoization (ESI) in positive and negative ion modes, the collection range was m/z 50-1 000.Agilent MassHunter Qualitative Analysis software was used to extract and match chromatographic peaks. Result::Combined with reference, related literature and database analysis, 95 compounds were identified by mass spectrometry information, including 41 flavonoids, 23 alkaloids, 12 lignans, 9 organic acids, and 10 other compounds. Conclusion::The chemical composition of Bufei Jianpi formula is complex, and the cracking rules of different components are different. Flavonoids are prone to deglycosylation, dehydration, Diels-Alder reaction (RDA) cleavage of the ring during lysis, and loss of some neutral molecules such as CO, CO2, CHO. Lignans has a substituent such as a hydroxyl group, a carbonyl group or a methoxy group on the benzene ring, and it is easy to obtain a fragment ion which loses H2O or CO. The basic structure of organic acids is a phenolic hydroxyl group-substituted aromatic ring, acrylic acid, fatty acid or the like, this kind of compound is easy to lose H2O and COOH in negative ion mode, and it is easy to break at the carbonyl to form fragment ions. This established method is rapid, sensitive and accurate, which can quickly identify the chemical constituents in Bufei Jianpi formula and provide evidences for clarifying efficacy material base of this formula.

7.
Article in Chinese | WPRIM | ID: wpr-849696

ABSTRACT

[Abstract] Objective To determine the prognostic value of thrombomodulin (TM) combined with thrombin-antithrombin complex (TAT) in patients with sepsis. Methods A retrospective analysis of the clinical data from 80 patients with sepsis who met the inclusion and exclusion criteria admitted to the 908th Hospital of Chinese PLA Logistical Support Force from May 2018 to July 2019. The conventional coagulation tests, thromboelastographic (TEG) parameters, TM, TAT, α2-plasmin inhibitor-plasmin complex (PIC, namely plasmin/alpha 2-antiplasmin complex, PAP) and tissue plasminogen activator-inhibitor-1 complex (t-PAIC) were collected within 2 hours at admission. According to the prognosis of 90 days, the patients were divided into survival group and death group and statistical analysis were performed. Results Compared with TM [11.7(8.8, 15.9) TU/ml] and TAT [11.3(7.0, 20.5) ng/ml] in the survival group, TM [20.2(14.1, 23.8) TU/ml] and TAT [17.7(11.8, 54.6) ng/ml] were significantly increased in the death group (P16.95 TU/ml combined with TAT>10.55 ng/ml. Conclusion TM combined with TAT can effectively judge the prognosis of sepsis patients and early identify sepsis related coagulation disorders.

8.
Article in Chinese | WPRIM | ID: wpr-828011

ABSTRACT

Forty-three annual Citrus aurantium grafted seedlings from Chongqing, Sichuan, Hunan, Jiangxi and other main producing areas were collected, and the plant height, rootstock diameter, scion diameter, root length, root diameter, lateral root number, root breadth, branch number, branch length, green leaf number, leaf length, leaf width, thorns and other indicators were measured. Through the K-cluster analysis of SPSS 19.0 software, the classification standards were obtained. Combined with the production practice, plant height, scion diameter and branch number were taken as the quality classification indexes of C. aurantium seedlings(annual grafted seedlings), and three classification standards were established. If it does not meet the three-level standard, it is unqualified seedling and cannot be used as seedling. It is suggested to use the first and second level seedlings in production.


Subject(s)
Citrus , Plant Leaves , Plant Roots , Seedlings
9.
Chinese Journal of Burns ; (6): 186-192, 2019.
Article in Chinese | WPRIM | ID: wpr-804886

ABSTRACT

Objective@#To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro.@*Methods@#The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference t test, and Bonferroni correction.@*Results@#(1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t3 h=4.891, 5.890, 4.928; t6 h=9.790, 6.750, 10.590; t9 h=6.948, 6.764, 5.049, P<0.05 or P<0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 μmol/L capsaicin group was increased (t=10.488, P<0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t=4.372, 3.026, P>0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 μmol/L capsaicin group and hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (t=15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P<0.05 or P<0.01), which of hypoxia+ 10.0 μmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t=4.365, P<0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t=6.216, P<0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t=3.489, P<0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t=0.545, P>0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t=12.550, 7.442, P<0.01).@*Conclusions@#TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.

10.
Chinese Journal of Burns ; (6): 169-178, 2019.
Article in Chinese | WPRIM | ID: wpr-804884

ABSTRACT

Objective@#To investigate the effect of human antigen R on lysosomal acidification during autophagy in mouse cardiomyocytes cultured in vitro.@*Methods@#The hearts of 20 C57BL/6 mice aged 1-2 days no matter male or female were isolated to culture primary cardiomyocytes which were used in the following experiments. (1) The cells were divided into 5 groups according to the random number table (the same grouping method below), i. e., normal control group and sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups. The cells in normal control group were routinely cultured for 54.0 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), and the cells in sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups were firstly regularly cultured for 53.5, 53.0, 51.0, 48.0 h and then cultured with replaced sugar-free serum-free medium for 0.5, 1.0, 3.0, and 6.0 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 Ⅱ (LC3Ⅱ), autophagy-related protein 5, and adenosine triphosphatase V1 region E1 subunit (ATP6V1E1) were detected by Western blotting. (2) The cells were divided into normal control group and sugar-free serum-free 3.0 h group. The cells in corresponding groups were treated the same as those in experiment (1), and the cell lysosomal acidification level was observed and detected under a laser scanning confocal microscope. (3) Two batches of cells were grouped and treated the same as those in experiment (1). The protein expression of human antigen R in the whole protein of cells of one batch and its protein expression in the cytoplasm and nucleus protein of cells of the other batch were detected by Western blotting. (4) The cells were divided into normal control group, simple control small interfering RNA (siRNA) group, simple human antigen R-siRNA1 (HuR-siRNA1) group, simple HuR-siRNA2 group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group. After 48 hours of regular culture, the cells in simple control siRNA group and sugar-free serum-free+ control siRNA group were transfected with negative control siRNA for 6 h, the cells in simple HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA1 group were transfected with HuR-siRNA1 for 6 h, and the cells in simple HuR-siRNA2 group and sugar-free serum-free+ HuR-siRNA2 group were transfected with HuR-siRNA2 for 6 h. Hereafter, the cells in these 8 groups were continuously cultured for 48 h with regular conditon, and then the cells in normal control group and each simple siRNA-treated group were replaced with DMEM/F12 medium, the cells in the other groups were replaced with sugar-free serum-free medium, and they were cultured for 3 h. The protein expression of human antigen R in the whole protein of cells was detected by Western blotting. (5) Two batches of cells were divided into sugar-free serum-free+ control siRNA group and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The distribution and expression of human antigen R in the cells of one batch were observed and detected by immunofluorescence method, and the lysosomal acidification level in the cells of the other batch was observed and detected under a laser scanning confocal microscope. (6) Three batches of cells were divided into sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, sugar-free serum-free+ HuR-siRNA1 group, and sugar-free serum-free+ HuR-siRNA2 group, and the cells in corresponding groups were treated the same as those in experiment (4). The protein expressions of cathepsin D in the whole protein of cells of one batch, human antigen R in the cytoplasm protein of cells of one batch, and ATP6V1E1 in the whole protein of cells of the other batch were detected by Western blotting. (7) The cells were divided into normal control group, sugar-free serum-free 3.0 h group, sugar-free serum-free+ control siRNA group, and sugar-free serum-free+ HuR-siRNA1 group, and the cells in corresponding groups were treated the same as those in experiment (4). The mRNA expression of ATP6V1E1 in cells was detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. The sample number of each experiment was 3. Data were processed with independent data t test, one-way analysis of variance, least significant difference t test, and Bonferroni correction.@*Results@#(1) Compared with those of normal control group, the protein expressions of LC3Ⅱ and ATP6V1E1 in the whole protein of cells of sugar-free serum-free 1.0, 3.0, and 6.0 h groups were significantly increased (t=12.16, 4.05, 4.82, 11.64, 3.29, 8.37, P<0.05 or P<0.01). Compared with that of normal control group, the protein expression of autophagy-related protein 5 in the whole protein of cells of sugar-free serum-free 0.5, 1.0, 3.0, and 6.0 h groups was significantly increased (t=6.88, 10.56, 5.76, 9.91, P<0.05 or P<0.01). (2) Compared with 1.03±0.08 of normal control group, the lysosomal acidification level in the cells of sugar-free serum-free 3.0 group (2.92±0.30) was significantly increased (t=6.01, P<0.01). (3) There was no statistically significant difference in the overall comparison of protein expression of human antigen R in the whole protein of cells among the 5 groups (F=1.09, P>0.05). Compared with that of normal control group, the protein expression of human antigen R in the cytoplasm protein of cells was significantly increased in sugar-free serum-free 1.0, 3.0, and 6.0 h groups (t=43.05, 11.07, 5.39, P<0.05 or P<0.01), while the protein expression of human antigen R in the nucleus protein of cells was significantly decreased in sugar-free serum-free 3.0 and 6.0 h groups (t=11.18, 12.71, P<0.01). (4) Compared with that of simple control siRNA group, the protein expression of human antigen R in the whole protein of cells of simple HuR-siRNA1 group and simple HuR-siRNA2 group was significantly decreased (t=4.82, 4.44, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the protein expression of human antigen R in the whole protein of cells of sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group was significantly decreased (t=4.39, 6.27, P<0.05). (5) Compared with those of sugar-free serum-free+ control siRNA group, the distribution of human antigen R in the cytoplasm of cells and its expression level were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group (t=10.13, P<0.01). Compared with 1.00±0.06 of sugar-free serum-free+ control siRNA group, the lysosomal acidification level (0.73±0.06) in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=3.28, P<0.01). (6) Compared with those of sugar-free serum-free+ control siRNA group, the protein expressions of cathepsin D in the whole protein of cells, human antigen R in the cytoplasm protein of cells, and ATP6V1E1 in the whole protein of cells were significantly decreased in sugar-free serum-free+ HuR-siRNA1 group and sugar-free serum-free+ HuR-siRNA2 group (t=4.16, 3.99, 4.81, 5.07, 11.68, 12.97, P<0.05 or P<0.01). (7) Compared with that of normal control group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free 3.0 h group was significantly increased (t=5.51, P<0.05). Compared with that of sugar-free serum-free+ control siRNA group, the mRNA expression of ATP6V1E1 in the cells of sugar-free serum-free+ HuR-siRNA1 group was significantly decreased (t=5.97, P<0.05).@*Conclusions@#After sugar-free serum-free treatment in vitro, the autophagy in mouse primary cardiomyocytes is activated, the lysosomal acidification is enhanced, and the expression of human antigen R in cytoplasm is increased. Human antigen R function is activated and involved in maintaining lysosomal acidification during autophagy in mouse cardiomyocytes.

11.
Chinese Journal of Burns ; (6): 116-124, 2019.
Article in Chinese | WPRIM | ID: wpr-804755

ABSTRACT

Objective@#To investigate the role of hexokinase Ⅱ in the changes of autophagic flow in cardiomyocytes of mice with ischemia-hypoxia in vitro.@*Methods@#The hearts of totally six male and female C57BL/6 mice aged from 1 to 2 days were isolated to culture primary cardiomyocytes which were used for the following experiments. (1) The cells were divided into 6 groups according to the random number table (the same grouping method below), i. e., normal control 3, 6, and 9 h groups and ischemia-hypoxia 3, 6, and 9 h groups, with 4 wells in each group. After being regularly cultured for 48 h with Dulbecco′s modified Eagle medium/nutrient mixture F12 (DMEM/F12) medium (the same regular culture condition below), the cells in normal control 3, 6, and 9 h groups were cultured with replaced fresh DMEM/F12 medium for 3, 6, and 9 h, respectively, and the cells in ischemia-hypoxia 3, 6, and 9 h groups were cultured with replaced sugar-free serum-free medium in the low-oxygen incubator with a volume fraction of 1% oxygen and a volume fraction of 5% carbon dioxide at 37 ℃ (the same hypoxic culture condition below) for 3, 6, and 9 h, respectively. Cell viability was measured by the cell counting kit 8 (CCK-8) method. (2) The cells were grouped and treated the same as those in experiment (1), with 1 well in each group. Western blotting was used to detect the protein expressions of microtubule-associated protein 1 light chain 3 Ⅰ (LC3Ⅰ), LC3Ⅱ, p62, and hexokinase Ⅱ. (3) The cells were divided into normal control group, simple ischemia-hypoxia 9 h group, and ischemia-hypoxia 9 h+ 2-deoxyglucose (2-DG) group, with 4 wells in each group. After a regular culture for 48 h, the cells in normal control group were cultured with replaced fresh DMEM/F12 medium for 9 h; the cells in simple ischemia-hypoxia 9 h group were replaced with sugar-free serum-free medium, and the cells in ischemia-hypoxia 9 h+ 2-DG group were replaced with sugar-free serum-free medium in which 2-DG was dissolved in a concentration of 10 mmol/L (20 μmol), and then they were cultured with hypoxia for 9 h. Cell viability was measured by CCK-8 method. (4) The cells were grouped and treated the same as those in experiment (3), with 1 well in each group. Western blotting was used to detect the protein expressions of LC3Ⅰ, LC3Ⅱ, and p62. (5) The cells were grouped and treated the same as those in experiment (3), with 2 wells in each group. Transmission electron microscope was used to observe autophagosomes/autolysosomes in cardiomyocytes. (6) The cells were divided into normal control group, simple ischemia-hypoxia 9 h group, ischemia-hypoxia 9 h+ hexosinase Ⅱ small interfering RNA1 (HK-ⅡsiRNA1) group, and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group, with 4 wells in each group. The cells in normal control group and simple ischemia-hypoxia 9 h group were regularly cultured for 48 h, and the cells in ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were respectively transfected with 200 nmol/L HK-ⅡsiRNA1 and HK-ⅡsiRNA2 and then also cultured for 48 h. The cells in normal control group were cultured with replaced fresh DMEM/F12 medium for 9 h, and the cells in simple ischemia-hypoxia 9 h group, ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group, and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were cultured with replaced sugar-free serum-free medium and hypoxia for 9 h. Cell viability was measured by CCK-8 method. (7) The cells were grouped and treated the same as those in experiment (6), with 1 well in each group. Western blotting was used to detect the protein expressions of LC3Ⅰ, LC3Ⅱ, p62, and hexokinase Ⅱ. Except for experiment (5), each experiment was repeated 3 times. Data were processed with one-way analysis of variance and lest significant difference t test, and Bonferroni correction.@*Results@#(1) The viabilities of cardiomyocytes in ischemia-hypoxia 3, 6, and 9 h groups were 0.450±0.022, 0.385±0.010, and 0.335±0.015, respectively, which were significantly lower than 0.662±0.026, 0.656±0.028, and 0.661±0.021 of the corresponding normal control 3, 6, and 9 h groups, respectively (t=6.21, 9.12, 12.48, P<0.01). (2) Compared with those of corresponding normal control 3, 6, and 9 h groups, the LC3Ⅱ/Ⅰ ratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of ischemia-hypoxia 3, 6, and 9 h groups were significantly increased (t3 h=16.15, 10.99, 5.30, t6 h=6.79, 10.42, 9.42, t9 h=15.76, 16.51, 7.20, P<0.05 or P<0.01). (3) The viability of cardiomyocytes in simple ischemia-hypoxia 9 h group was 0.353±0.022, which was significantly lower than 0.673±0.027 of normal control group (t=9.29, P<0.01). The viability of cardiomyocytes in ischemia-hypoxia 9 h+ 2-DG group was 0.472±0.025, which was significantly higher than that of simple ischemia-hypoxia 9 h group (t=3.60, P<0.05). (4) Compared with those of normal control group, the LC3Ⅱ/Ⅰ ratio and protein expression of p62 in cardiomyocytes of simple ischemia-hypoxia 9 h group were significantly increased (t=9.45, 8.40, P<0.01). Compared with those of simple ischemia-hypoxia 9 h group, the LC3Ⅱ/Ⅰratio and protein expression of p62 in cardiomyocytes of ischemia-hypoxia 9 h+ 2-DG group were significantly decreased (t=4.39, 4.74, P<0.05). (5) In cardiomyocytes of normal control group, only single autophagosome/autolysosome with bilayer membrane structure was observed. Compared with that of normal control group, the number of autophagosome/autolysosome with bilayer membrane structure in cardiomyocytes of simple ischemia-hypoxia 9 h group was increased significantly. Compared with that of simple ischemia-hypoxia 9 h group, the number of autophagosome/autolysosome with bilayer membrane structure in cardiomyocytes of ischemia-hypoxia 9 h+ 2-DG group was significantly decreased. (6) The viability of cardiomyocytes in simple ischemia-hypoxia 9 h group was 0.358±0.023, which was significantly lower than 0.673±0.026 in normal control group (t=9.12, P<0.01). The viabilities of cardiomyocytes in ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were 0.487±0.027 and 0.493±0.022, respectively, which were significantly higher than the viability in simple ischemia-hypoxia 9 h group (t=3.63, 4.28, P<0.05). (7) Compared with those of normal control group, the LC3Ⅱ/Ⅰratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of simple ischemia-hypoxia 9 h group were significantly increased (t=6.08, 6.31, 4.83, P<0.05 or P<0.01). Compared with those of simple ischemia-hypoxia 9 h group, the LC3Ⅱ/Ⅰ ratio and protein expressions of p62 and hexokinase Ⅱ in cardiomyocytes of ischemia-hypoxia 9 h+ HK-ⅡsiRNA1 group and ischemia-hypoxia 9 h+ HK-ⅡsiRNA2 group were significantly decreased (t=5.10, 7.76, 15.33, 4.17, 8.42, 12.11, P<0.05 or P<0.01).@*Conclusions@#Ischemia-hypoxia upregulates the expression level of hexokinase Ⅱ protein in mouse cardiomyocytes cultured in vitro, which decreases the viability of cardiomyocytes by impairing autophagic flow. To inhibit the activity of hexokinase Ⅱ or its expression can alleviate the ischemia-hypoxia damage of cardiomyocytes.

12.
Chinese Journal of Microsurgery ; (6): 268-271, 2019.
Article in Chinese | WPRIM | ID: wpr-756326

ABSTRACT

Objective To identify the routes and branches of the proper plantar digital nerves(PPDN) in the medial of the great toe and its adjoining relationship among the surrounding fascia tissues and organs,which was expected to provide accurate localization of the nerve impingement and possible relevant of anatomical basis for the treatment of nerve entrapment in clinical utility.Methods From December,2016 to January,2019,a total of 54 formalin fixed feet were collected.Fifty of them were performed conventional anatomical procedure,the other 4 were prepared with sectional anatomical technique.The seats and branches of the PPDN in the medial of the great toe were observed;The width and thickness of the nerve were measured at the first metatarpophalangeal joint(FMPJ),along with its proximal and distal sides 0.5 cm.The origin and origin of fascia were observed by foot dissection.Masson staining was used to observe the tissue changes of the nerves in the FMPJ.Results The PPDN of the medial great toe run between the flexor pollicis longus tendon and the abductor pollicis tendon at the proximal,issued (4.21±0.12) final branches.And governed the sensation of the medial half of the great toe.The width of the nerve at the FMPJ was (3.50±0.09) mm,which was significantly increased compared with that of the near [(1.58±0.04) mm] and far [(1.56± 0.03) mm] from the joint.The difference was statistically significant (P<0.05);The thickness of the nerve in the proximal segment was (0.83±0.04) mm,and that in the distal segment was (0.82±0.03) mm.Compared with that in the FMPJ [(0.67±0.02) mm],the difference was statistically significant (P<0.05).A deep fascia was observed on the superficial surface of the PPDN at medial great toe,which was stretched between the tendon sheath of the flexor pollicis longus tendon and the tendon of the abductor pollicis muscle.Masson staining showed obvious proliferation of nerve outer mem brane fibers at the metatarpophalangeal joint,the number of nerve fiber bundles increased,and obvious thickening of nerve fiber bundles and nerve fascia.Conclusion Long-term compression can lead to thickening of the epineurium and perineurium,and the superficial fascia is an important factor of thumb pain and numbness caused by the compression of the PPDN at medial of the great toe.

13.
Article in Chinese | WPRIM | ID: wpr-745901

ABSTRACT

Clinical data of a 33-year-old rheumatoid arthritis patient accompanied with pregnancy and lactation-associated osteoporosis(PLO) was retrospectively reviewed.The patient had two pregnancies within three years,and bone pain occurred during lactation of the second pregnancy after 2 months of delivery.Dual-energy X-ray (DEXA) showed decreased bone density,while thoracolumbar spine and pelvic X-ray film suggested osteoporosis.Her rheumatoid arthritis was stable and previous treatment did not include glucocorticoids.Other causes of metabolic bone diseases were excluded and the diagnosis of PLO was confirmed.The breastfeeding was stopped;calcium,active vitamin D,alendronate sodium were administrated,and the treatment was effective.Domestic and international literature from January 2000 to January 2019 was searched and no cases of PLO accompanied with rheumatoid arthritis were reported.There were only 100 cases of PLO retrieved in the literature,including 19 cases of PLO in China reported by four articles.It is suggested that PLO should be considered in RA patients presenting with arthragia during the lactation,in addition to relapse of disease.Awareness of PLO should be increased in physicians,especially rheumatologists,and anti-osteoporosis therapy is effective in patients with PLO.

14.
Journal of Medical Postgraduates ; (12): 845-849, 2019.
Article in Chinese | WPRIM | ID: wpr-818334

ABSTRACT

Objective The platelet function changes are closely related to the prognosis of trauma patients and the occurrence of coagulopathy. The purpose of this paper is to investigate the clinical value of platelet function changes in trauma patients for prognosis judgment. Methods The clinical data of 94 trauma patients admitted to the Department of Critical Care Medicine, 908th Hospital from July 2017 to February 2019 were retrospectively analyzed. According to the 90-day prognosis of patients, the patients were divided into survival group (n=80) and death group (n=14) to compare the traditional coagulation function indexes, including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), thrombin time (TT), fibrinogen degradation product (FDP), D-dimer, antithrombin II (I ATIII), thromboelastogram (TEG) index [coagulation reaction time (R), clot formation rate (K), clot formation kinetics (α angle), maximum clot strength (MA), etc.] and platelet aggregation function index [arachidonic acid (AA) platelet aggregation rate and adenosine diphosphate (ADP) Platelet aggregation rate]. The data was analyzed by receiver operating characteristic (ROC) curve analysis and Kaplan-Meier analysis. Results Compared with the survival group, the APPT, R value and K value prolonged significantly in the death group (P<0.05). However, the MA value,AA-induced and ADP-induced platelet aggregation decreased significantly in the death group (P<0.05). The ROC curve analysis showed that when the MA cut-off value was 42.05mm, the sensitivity, specificity, positive predictive value and negative predictive value were 83.8%, 71.4%, 58.3% and 94.2% respectively. When the cut-off value of AA platelet aggregation rate was 36.6%, the sensitivity, specificity, positive predictive value and negative predictive value were 57.5%, 85.7%, 75.5% and 93.8% respectively. When the cut-off value of ADP platelet aggregation rate was 29.3%, the sensitivity, specificity, positive predictive value and negative predictive value were 70%, 64.3%, 72.7% and 91.8% respectively. The death risk of patients with AA-induced aggregation rate < 36.6% was 4.37 times that of the patients with AA-induced platelet aggregation rate ≥ 36.6% (95% CI: 1.34 to 10.98). The death risk of patients with ADP-induced aggregation rate < 29.3% was 3.674 times that of the patients with ADP-induced platelet aggregation rate ≥ 29.3% (95%CI:1.385~ 12.880). The death risk of trauma patients with MA < 42.05 mm was 9.759 times that of the patients with MA ≥ 42.05 mm (95% CI: 6.674 ~ 89.87). Conclusion The platelet function of trauma patients can be significantly impaired. When the MA, AA platelet aggregation rate and ADP platelet aggregation rate are lower, the mortality rate of trauma patients becomes higher. The platelet function index of MA, AA and ADP can be used to determine the prognosis of trauma patients.

15.
Journal of Medical Postgraduates ; (12): 196-200, 2019.
Article in Chinese | WPRIM | ID: wpr-818211

ABSTRACT

The clinical treatment of sepsis has always been a problem. Sepsis is characterized by high morbidity, high mortality and high treatment costs.Platelets were previously thought to play a major role in hemostasis and thrombosis.In recent years, it has been found that platelets are very important in inflammation and immune regulation as well as in hemostasis and thrombosis. Antiplatelet therapy can reduce release of inflammation and immunomodulatory mediators by attenuating platelet activation, thereby alleviating the inflammatory response in sepsis. This paper reviews the mechanism of platelet involvement in sepsis and the clinical progress of antiplatelet drugs in managing sepsis.

16.
Journal of Medical Postgraduates ; (12): 1184-1188, 2019.
Article in Chinese | WPRIM | ID: wpr-818164

ABSTRACT

Objective According to the cell-based coagulation theory, antithrombin complex (TAT) reflecting the activation of coagulation system, plasmin-α2 anti-plasmin complex (PIC) reflecting the activation of fibrinolytic system, thrombomodulin (TM) and tissue plasminogen activator-plasminogen activator inhibitor-1 complex (t-PAIC) reflecting vascular endothelial function were selected to explore their diagnostic values for disseminated intravascular coagulation. Methods A prospective study was conducted on 154 patients in the Department of Critical Care Medicine of the 908th Hospital from May to December 2018. The subjects were divided into non-overt DIC group (n=134) and overt DIC group (n=20) according to the diagnostic criteria of International Thrombus and Hemostatic Association. The differences among groups of TM, t-PAIC, TAT and PIC were compared along with statistical analysis. Results Compared with TM [10.5 (8.0~14.3) TU/mL], TAT [9.6 (4.9~21.8) ng/mL], PIC [1.253 (0.789~2.802) μg/mL] and t-PAIC [ 11.2 (7.1~22.1) ng/mL] in non-overt DIC group, TM [16.8 (11.8~21.5) TU/mL], TAT [33.6 (10.3~120.0) ng/mL], PIC [4.080 (0.814~8.651) μg/mL] and t-PAIC [19.4 (10.0~30.1)ng/mL] ) in overt DIC group were significantly increased (P<0.05). The area under the curve of TM>14.85 TU/mL combined with TAT>23.05 ng/mL as the standard diagnostic overt DIC was 0.835 (P=0.000), and the sensitivity, specificity, positive predictive value and negative predictive value were 0.85, 0.761, 0.592, 0.925 respectively. Conclusion TM combined with TAT has a higher diagnostic efficacy for overt DIC.

17.
Article in Chinese | WPRIM | ID: wpr-818117

ABSTRACT

Objective Bloodstream infection (BSI)-induced sepsis may cause severe coagulation dysfunction. In this study, we analyzed the characteristics of coagulation dysfunction in different phases of BSI-induced sepsis.Methods We retrospectively analyzed the clinical data on 43 cases of BSI treated in our Department of Critical Care Medicine from January 2016 to September 2018. According to the Diagnostic Criteria for Sepsis 3.0, we divided the patients into a sepsis group and a septic shock group, compared the traditional indexes of coagulation function and parameters of thromboelastography (TEG) between the two groups, and analyzed the obtained data with the ROC curves.Results Compared with the sepsis group, the septic shock group showed significantly prolonged prothrombin time (PT) (13.7 \[12.5-17.4] vs 16.7 \[15.0-20.9\] s, P0.05).Conclusion Coagulation dysfunction in BSI-induced sepsis is characterized by elevated values of DD and FDP, and in case of septic shock, it features low coagulation of clotting factors (R≥8.5 min) and fibrin function (K≥2.65 min). The patient with septic shock may experience significantly reduced PLT with basically normal platelet function.

18.
Article in Chinese | WPRIM | ID: wpr-775887

ABSTRACT

Needle retention is an important step in the acupuncture procedure. How to optimize scientifically the duration of needle retention according to individual case has been considered in the medical circle. In this paper, by collecting the literatures on needle retention from the early dynasty to the contemporary time, the evolution of the needle retention from a short duration to a long one with the productivity improvement was elaborated. On the base of the views of the medical scholars of all dynasties, it was concluded that the ultimate purpose of needle retention is to improve the effects of acupuncture on the premise of ensuring the safety of acupuncture. Hence, the clinical physician should optimize the duration of needle retention cautiously in compliance with the tolerance of patient so as to save the time cost of both physician and patient, avoid the occurrence of tolerable effect of acupuncture and reduce the potential safety hazard of acupuncture induced by the long duration of needle retention.


Subject(s)
Acupuncture Therapy , Humans , Moxibustion , Needles , Physicians , Time Factors
19.
Gut and Liver ; : 569-575, 2019.
Article in English | WPRIM | ID: wpr-763869

ABSTRACT

BACKGROUND/AIMS: Cholangiocytes are capable of reabsorbing bile salts from bile, but the pathophysiological significance of this process is unclear. To this end, we detected the expression and distribution of bile acid transport proteins in cholangiocytes from normal rat liver and analyzed the possible pathophysiological significance. METHODS: Bile duct tissues of Sprague-Dawley rats were isolated by enzymatic digestion and mechanical isolation, and then divided into large and small bile duct tissues. Immunohistochemistry, real-time polymerase chain reaction and Western blotting were used to determine the expression of the apical sodium-dependent bile acid transporter (ASBT), ileal bile acid binding protein (IBABP), and basolateral organic solute transporter α (Ostα) in the biliary tract system of rats. Differences in the expression and distribution of these proteins were analyzed. RESULTS: In cholangiocytes, ASBT and IBABP were mainly expressed in cholangiocytes of the large bile ducts, in which the expression of both was significantly higher than that in the small ducts (p0.05). CONCLUSIONS: Bile acid transporters are expressed and heterogeneously distributed in rat bile ducts, indicating that bile acid reabsorption by cholangiocytes might mainly occur in the large bile ducts. These findings may help explore the physiology of bile ducts and the pathogenesis of various cholangiopathies.


Subject(s)
Animals , Bile Acids and Salts , Bile Ducts , Bile , Biliary Tract , Blotting, Western , Carrier Proteins , Digestion , Immunohistochemistry , Liver , Physiology , Population Characteristics , Rats , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction
20.
Article in Chinese | WPRIM | ID: wpr-803367

ABSTRACT

Objective@#To explore the A wave value in neuroelectrophysiological subtype of Guillain-Barré syndrome(GBS)and the clinical severity and short-term prognosis of acute inflammatory demyelinating polyradiculoneuropathy(AIDP).@*Methods@#From March 2014 to March 2017, a total of 56 children with GBS at Department of Neurology of Wuhan Children′s Hospital Affiliated to Tongji Medical College, Huazhong University of Science & Technology were enrolled.The patients were divided into AIDP subtype(40 cases) and axonal GBS subtype(16 cases) according to the results of electrophysiological examination.According to whether there was existence of A wave or not, the GBS children were divided into 2 groups.The first group was the A wave in GBS group(18 cases), and the second group was non-A wave in GBS group(38 cases). In order to explore classification value for GBS with A wave, clinical data including age, gender, history of prodromal infection, cranial nerve dysfunction, autonomic nerve involvement and conduction blocks were analyzed.To explore A wave value in clinical severity and short-term prognosis of AIDP, the age, gender, clinical severity, conduction blocks, short-term prognosis of the 2 groups were analyzed in A wave with AIDP (18 cases) and non-A wave with AIDP(22 cases).@*Results@#Compared with non-A wave GBS patients, A wave GBS patients had more conduction blocks(10 cases vs.2 cases, χ2=18.021, P=0.000). Age, sex, precedent infections, cranial nerve involvement, autonomic nerve involvement were not significantly statistically different(all P>0.05). A wave was only seen in AIDP subtype(18 cases), and the percentage of A wave in AIDP was 45%(18/40 cases). There was no A wave in axonal GBS.Compared with non-A wave in AIDP, A wave in AIDP patients had more conduction blocks(10 cases vs.2 cases, χ2=9.924, P=0.002), poorer clinical motor function[(3.39±1.09) scores vs.(2.50±1.01) scores, t=2.667, P=0.011]and short-term prognosis[(2.06±0.64) scores vs.(1.55±0.60) scores, t=2.607, P=0.013].@*Conclusions@#A wave is correlated with demyelination subtype in children′s Guillain-Barré syndrome and poor short-term prognosis in AIDP.

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