Protective effect of metformin on pulmonary fibrosis caused by paraquat through activating AMP-activated protein kinase pathway / 中华危重病急救医学

OBJECTIVE@#To observe whether metformin (MET) inhibits transforming growth factor-β1 (TGF-β1)/Smad3 signaling pathway by activating adenosine activated protein kinase (AMPK), so as to alleviate the pulmonary fibrosis caused by paraquat (PQ) poisoning in mice.@*METHODS@#Male C57BL/6J mice were randomly divided into the Control group, PQ poisoning model group (PQ group), MET intervention group (PQ+MET group), AMPK agonist group (PQ+AICAR group), and AMPK inhibitor group (PQ+MET+CC group), according to a random number table method. A mouse model of PQ poisoning was established by one-time peritoneal injection of 1 mL PQ solution (20 mg/kg). The Control group was injected with the same volume of normal saline. After 2 hours of modeling, the PQ+MET group was given 2 mL of 200 mg/kg MET solution by gavage, the PQ+AICAR group was given 2 mL of 200 mg/kg AICAR solution by intraperitoneal injection, the PQ+MET+CC group was given 2 mL of 200 mg/kg MET solution by gavage and then 1 mL complex C (CC) solution (20 mg/kg) was intraperitoneally injected, the Control group and PQ group were given 2 mL of normal saline by gavage. The intervention was given once a day for 21 consecutive days. The 21-day survival rate of ten mice in each group was calculated, and the lung tissues of remaining mice were collected at 21 days after modeling. The pathological changes of lung tissues were observed under light microscope after hematoxylin-eosin (HE) staining and Masson staining, and the degree of pulmonary fibrosis was evaluated by Ashcroft score. The content of hydroxyproline in lung tissue and oxidative stress indicators such as malondialdehyde (MDA) and superoxide dismutase (SOD) were detected. The protein expressions of E-cadherin, α-smooth muscle actin (α-SMA), phosphorylated AMPK (p-AMPK), TGF-β1 and phosphorylated Smad3 (p-Smad3) in lung tissue were detected by Western blotting.@*RESULTS@#Compared with the Control group, the 21 days survival rate was significantly reduced, lung fibrosis and Ashcroft score were significantly increased in PQ group. In addition, the content of hydroxyproline, MDA and the protein expressions of α-SMA, TGF-β1 and p-Smad3 in lung tissue were significantly increased, while the activity of SOD and the protein expressions of E-cadherin and p-AMPK were significantly decreased in PQ group. Compared with the PQ group, the 21 days survival rates of mice were significantly improved in the PQ+MET group and PQ+AICAR group (70%, 60% vs. 20%, both P < 0.05). The degree of pulmonary fibrosis and the Ashcroft score were significantly reduced (1.50±0.55, 2.00±0.63 vs. 6.67±0.52, both P < 0.05). The content of hydroxyproline and MDA in lung tissue, as well as α-SMA, TGF-β1 and p-Smad3 protein expressions were significantly reduced [hydroxyproline (mg/L): 2.03±0.11, 3.00±0.85 vs. 4.92±0.65, MDA (kU/g): 2.06±1.48, 2.10±1.80 vs. 4.06±1.33, α-SMA/GAPDH: 0.23±0.06, 0.16±0.06 vs. 1.00±0.09, TGF-β1/GAPDH: 0.28±0.03, 0.53±0.05 vs. 0.92±0.06 p-Smad3/GAPDH: 0.52±0.04, 0.69±0.06 vs. 1.11±0.10, all P < 0.05], SOD activity and the protein expressions of E-cadherin and p-AMPK were significantly increased [SOD (μmol/g): 39.76±1.35, 33.03±1.28 vs. 20.08±1.79, E-cadherin/GAPDH: 0.91±0.08, 0.72±0.08 vs. 0.26±0.04, p-AMPK/GAPDH: 0.62±0.04, 0.60±0.01 vs. 0.20±0.04, all P < 0.05]. However, these protective effects of MET were inhibited by the addition of AMPK inhibitor CC solution.@*CONCLUSIONS@#MET can effectively alleviate the degree of pulmonary fibrosis in mice poisoned with PQ, and its mechanism may be related to the activation of AMPK and inhibition of TGF-β1/Smad3 signaling pathway, which can be inhibited by AMPK inhibitor CC.

Effects of long-term high-fat diet on bone mineral density and intestinal flora / 中华内分泌代谢杂志

Objective:To investigate the effects of long-term high-fat diet on bone mineral density and intestinal flora in mice.Methods:Sixteen male C57BL/6 mice were randomly divided into control group (NC group) and high-fat group (HF group). After 24 weeks of high-fat feeding, biochemical indicators such as blood glucose and blood lipids were detected, bilateral femurs were taken and bone microstructure was analyzed with micro-computered tomography (micro-CT), and changes of intestinal microbial composition and proportion were revealed using 16S rDNA sequencing technology.Results:Compared with the control group, the serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in HF group were significantly increased. Micro-CT uncovered that the bone mineral density (Tb.BMD), bone volume fraction (BV/TV) and the number of trabecular bone (Tb.N) decreased, yet structural model index (SMI) and the trabecular fraction (Tb.Sp) increased in the HF group mice. The gut microbiota 16S rDNA sequence analysis showed that the proportion of Proteobacter was significantly increased and the proportions of pachycete, warty microbacterius, and actinomycete were reduced in HF group at the phyla level. The proportion of Bacteroidetes S24-7_norank in the NC group was significantly higher than that in the HF group, and the multilevel discriminant analysis of species differences (LEfSe) identified that the difference was significant, yet the proportion of Bacteroides, Pseudo-Prevotella, Desulfovibrio, Altobacter, and Helicobacter in the HF group were higher than those in the NC group, which were significant differences in Altobacter and Helicobacter at genus level.Conclusion:Long-term high-fat feeding can cause the destruction of femoral trabecular structure, decrease in the number of trabeculus bones, and bone mineral density in C57BL/6 mice. It also leads to significant changes in the composition and proportion of the intestinal flora.

Autosomal recessive hypophosphatemic rickets 1 caused by DMP1 mutation: one pedigree study / 中华内分泌代谢杂志

In the present study, the clinical features of a patient with autosomal recessive hypophosphatemic rickets 1 caused by dentin matrix protein 1(DMP1)gene mutation and her family members were investigated. DMP1 gene from peripheral blood was sequenced by Sanger sequencing, and the known mutation was verified among her family members and 250 healthy populations. The proband was a 42-year-old female with bone deformity of both lower limbs, bone pain, and short stature. The results of X-rays and laboratory examination were consistent with the hypophosphatemic rickets reported before. A homozygous mutation(c.2T> C)in DMP1 was identified by Sanger sequencing in the proband, her son and daughter were heterozygous for c. 2T> C.

Progress in the study of pulmonary fibrosis signaling pathways caused by paraquat poisoning / 中华危重病急救医学

Paraquat is a quaternary ammonium herbicide, which can be distributed in lung, liver, kidney, heart, brain and other organs through blood circulation, leading to multiple organ failure, especially lung injury. Due to the lack of effective treatment methods and specific antidotes, the prognosis of most patients with paraquat poisoning is very poor. The treatment of paraquat poisoning was a big problem for emergency doctors. Previous studies have found that pulmonary fibrosis caused by paraquat poisoning is closely related to a variety of pathological processes, such as oxidative stress, inflammatory reaction, mitochondrial damage, imbalance of extracellular matrixproduction (ECM) and degradation, which involve the activation or inhibition of various signaling pathways. In recent years, many researchers focused on clarifying the mechanism of paraquat induced pulmonary fibrosis, and some signaling pathways related to paraquat poisoning leading to pulmonary fibrosis have been found. A large number of studies have found that adenosine monophosphate activated protein kinase (AMPK) related signaling pathway, transforming growth factor-β/Smad (TGF-β/Smad)signaling pathway, mitogen-activated protein kinase (MAPK) related signaling pathway, Ras homolog gene/Rho associated kinases (Ras/ROCK) and Wnt/β-catenin signal pathways are closely related to paraquat induced pulmonary fibrosis. In this paper, we reviewed signaling pathways related to paraquat induced pulmonary fibrosis, in order to provide more ideas for the clinical treatment of paraquat induced pulmonary fibrosis.

Cross-cultural adaptation and reliability and validity testing of the Chinese version of Edmonton Frail Scale / 中国实用护理杂志

Objective:To translate the English version of Edmonton Frail Scale into Chinese version and to test its reliability and validity.Methods:The translation and cross-cultural adaptation of the English version of Edmonton Frail Scale were generated by Beaton's translation and back-translation method. From April to August of 2019, at the department of geratology of the First affiliated Hospital of China Medical University in Shenyang, a sample of 303 hospitalized elderly adults who met the inclusion and exclusion criteria were selected via convenience sampling method to test its psychometric properties.Results:The scale level content validity index and item level content validity index of the Chinese version of the Edmonton Frail Scale were both 1.0. The criterion validity between the Chinese version of Edmonton Frail Scale and the Tilburg Frailty Indicator was 0.723 ( P<0.01), confirmed by Pearson correlation coefficients. Using the Tilburg Frailty Indicator as an external criterion, the Edmonton Frail Scale showed satisfactory diagnostic accuracy for frailty (area under the curve=0.924). The optimal cut-point for frailty was 6 (sensitivity: 77.6%, specificity: 94.7%). More frail individuals were recognized by the Chinese version of Edmonton Frail Scale among older and female participants than their counterparts( P<0.05). The Cronbach'α of the Chinese version of Edmonton Frail Scale was 0.599, the test-retest reliability within a 7-day interval of the scale was 0.822. Conclusions:The Chinese version of Edmonton Frail Scale presents good validity and reliability and can apply to Chinese hospitalized elderly people.

Proximal symphalangism caused by NOG gene novel mutation: A family report / 中华内分泌代谢杂志

Proximal symphalangism is a rare hereditary bone disease caused by NOG or GDF5 gene mutations, of which NOG gene mutations account for the majority. A family of SYM1 was reported. Patient was a man with proximal interphalangeal joint stiffness of bilateral fingers for more than 20 years. Combined with laboratory and imaging examinations, the patient was diagnosed with proximal symphalangism. 4 other subjects in this family are affected. The detection of NOG gene mutations of the proband and his mother and son showed that there were heterozygous missense mutations in exon 1, c.667C>T, resulting in p. Pro223Ser. The pathogenesis, clinical and imaging manifestations of SYM1 were reviewed in combination with literature to improve clinicians′ understanding of the disease.

Progress in diagnosis and treatment of Hashimoto's thyroiditis / 中华全科医师杂志

Hashimoto's thyroiditis is the most common autoimmune thyroid disease and also the primary cause of hypothyroidism.The disease is characterized by long course, slow development, complicated clinical presentation and coexistence with a variety of thyroid diseases.Misdiagnosis and missed diagnosis rates are high,therefore close attention should be attached clinically.

Effect of low-dose or standard-dose conjugated equine estrogen combined with different progesterone on bone density in menopause syndrome women / 中华妇产科杂志

Objective To explore the effect of low-dose or standard-dose conjugated equine estrogen (CEE)combined with natural progesterone or dydrogesterone on bone density in menopause syndrome women.Methods Totally 123 patients with menopause syndrome were recruited and randomly assigned to 3 treatment groups: group A(low-dose CEE+progesterone), group B(standard-dose CEE+progesterone), group C(standard-dose CEE+dydrogesterone). Using continuous sequential regimen, the duration of intervention was 12 cycles.The bone mineral density of lumbar 2-4 and neck of femur,the bone metabolic markers, the level of FSH and estradiol were examined just before the drug administration and 12 months after the beginning of experiment. Results There were 107 cases completed the one year trial.(1)Bone density:after 12 cycles of treatment,there was no significant change in bone density in group A(P>0.05);lumbar vertebrae of group B and C increased significantly,at 3.0% and 2.1%respectively(all P<0.05).The bone density of left femoral neck of group C significantly increased by 2.9%(P=0.029). There was no significant difference among the treatment groups at the beginning of experiment(P>0.05).(2)Bone metabolic markers: after 12 cycles of treatment, the levels of calcium, phosphorus, alkaline phosphatase, Ca/Cr decreased significantly,the difference were statistically significant(all P<0.05).There was no significant difference among the treatment groups at the beginning of experiment(P>0.05).(3)Levels of FSH and estradiol:after 12 cycles of treatment,the levels of FSH in three groups were decreased significantly(all P<0.01). The levels of estradiol in three groups were increased significantly(all P<0.01). There was no significant difference among the treatment groups at the beginning of experiment(P>0.05). Conclusions Both low-dose and standard-dose menopause hormone therapy(MHT)could elevate the level of estradiol, reduce bone turnover, prevent bone loss of postmenopausal women effectively. The standard dose of MHT could also increase the density of vertebrae and femoral neck,and generate more clinical benefits.

Influence of CYP3A5*3 Genetic Polymorphism on Blood Concentration of Tacrolimus and Renal Function in Renal Transplant Recipients during the Stable Period / 中国药房

OBJECTIVE:To investigate the influence of CYP3A5*3 (rs776746) genetic polymorphism on blood concentration of tacmlimus (TAC) and renal function in renal transplant recipients during the stable period.METHODS:A total of 98 renal transplant recipients during the stable period receiving TAC-based triple anti-rejection scheme (TAC + sodium mycophenol +predrnisone acetate) after surgery and regular follow-up were selected from our hospital during Jan.1995-Dec.2014.The follow-up information during Jan.-Dec.2016 was also collected.Trough concentration of TAC in renal transplant recipients was determined by chemiluminescence microparticle immuno assay.Standard blood concentration (C/D) was calculated after corrected with body weight and daily dose.Scr level was detected with dry chemistry method.CYP3A5*3 genotype was detected by PCR-RFLP and direct sequencing.The relationship of CYP3A5*3 genetic polymorphism with TAC C/D value and Scr level was determined by Kruskal Wallis H or Mann-Whitney U assay.RESULTS:Among 98 renal transplant recipients,there were 9 cases of CYP3A5*3 *1/*1(AA) genotype,37 cases of *1/*3 (AG) genotype and 52 cases of *3/*3 (GG)genotype.The gene frequencies were 9.18％,37.76％,53.06％,which were all in line with Hardy-Weinberg equilibrium (P＞0.05).There was no statistical significance in trough concentration of TAC among different genotypes (P＞0.05).There was statistical significance in TAC dose and C/D value among different genotypes (P＞0.05).TAC dose of CYP3A5*3 *3/*3 genotype recipients was significantly lower than those of *1/*3 and *1/*1 genotype recipients;that of *1/*3 genotype recipients was significantly lower than that of *1/*1 genotype recipients.C/D value of *3/*3 genotype recipients was significantly higher than those of *1/*3 and *1/*1 genotype recipients;that of *1/*3 genotype recipients was significantly higher than that of *1/*1 genotype recipients,with statistical significance (P＜0.05).There was no statistical significance in Scr levels among different genotypes (P＞0.05).CONCLUSIONS:CYP3A5*3 genetic polymorphism significantly influences blood concentration of TAC in renal transplant recipients during the stable period,and *3 allele carriers have higher C/D values and need smaller TAC daily dose.CYP3AS*3 genetic polymorphism may be not associated with Scr level.

Determination of sinoporphyrin sodium in tumor-bearing mouse plasma by HPLC method / 药学实践杂志

Objective To establish a HPLC method for the assay of sinoporphyrin sodium (DVDMS) in tumor-bearing mouse plasma and to study its pharmacokinetics .Methods The column was Waters XBridge C18 (3 .0 mm × 100 mm ,3 .5μm) . Gradient elution was applied with mobile phase A as the mixture of acetonitrile-methanol (20:80) and B as the aqueous solu-tion of 1％ acetic acid and 0 .1％ triethylamine at flow rate 0 .7 ml/min .The detection wavelength was 380 nm .DVDMS was administrated to tumor-bearing mice by tail vein injection .The blood samples were collected at designated time and centrifuged for plasma .DVDMS in plasma samples were extracted by protein precipitation and analyzed by the HPLC method mentioned a-bove .Pharmacokinetic parameters were calculated by DAS 2 .0 with statistical moment analysis .Results DVDMS showed good linearity within the ranges of 70 .8-14160 ng/ml (r=0 .9998) .The main pharmacokinetic parameters were calculated as follows :cmax = (24127 .59 ± 1415 .23) ng/ml ,tmax =0 .083 h ,t1/2 = (9 .59 ± 1 .25) h ,MRT0-∞ = (11 .77 ± 1 .73) h ,AUC0-∞ =(34775 .83 ± 6185 .43) h · ng/ml .Conclusion This HPLC method is sensitive ,rapid and accurate ,which can be used for a-nalysis and research of DVDMS in plasma samples of tumor-bearing mice .

The value of post processing technique of MSCT in the diagnosis of bile duct stones / 实用放射学杂志

Objective To investigate the value of post processing technique of MSCT in the diagnosis of bile duct stones.Methods 89 cases with high density bile stones were collected.All of the images were reconstructed by using surface reconstruction(CPR),multiplanar reconstruction(MPR),volume reconstruction(VR), to clearly show the location, size, number and shape of bile duct stones, and provide accurate image information for clinic.Results 396 cases of bile duct stones were detected in all of the 89 patients,after treatment,the reconstructed image of could accurately show the location,size,number and shape of stones.Conclusion Post-processing technique of MSCT can provide accurate image information for the diagnosis of the the biliary stone,and improve the effectiveness and safety of the operation.

Clinical study of anti-Mullerian hormone in prediction of ovarian aging / 中华妇产科杂志

Objective To explore the serum anti-Mullerian hormone (AMH) level in women of childbearing age with normal menstrual cycles. Methods A total of 1 423 women with regular menstrual cycles were selected and divided into 5 groups according to their ages, i.e.≤25, 26-30, 31-35, 36-40,≥41 years. Their serum levels of AMH were measured, and the relationship between AMH and age was analyzed. Results The serum AMH levels of 5 groups according to ages (≤25, 26-30, 31-35, 36-40, ≥41 years) were 3.62, 3.10, 2.27, 1.07, 0.45μg/L, respectively. The comparison of serum AMH levels in different age groups had significant difference (P<0.01). Serum AMH level declined with increasing age,and dropped significantly after 36. The serum AMH level and age showed a negative correlation with significant difference (r=-0.374, P<0.01). Quadratic regression of logAMH proximally reflected the relationship between AMH and age. Conclusion AMH determination for women of childbearing age could provide reference for the evaluation of ovarian function.

Gene expression profiles of toxicated BALB/c mouse liver exposed to tetracycline / 第三军医大学学报

Objective To study the molecular mechanism of the toxic effect for tetracycline acting on liver using toxicological microarray.Methods Utilizing the mouse toxicological microarray and animal model of tetracycline injuring BALB/c mouse liver,both of which were established by our department,we observed the gene expression profiles in different dose groups and at different time points after tetracycline treatment,and primarily analysed the function of these differentially expressed genes using Gene Ontology Consortium analysis system and hierarchical method.Results Multiple differentially expressed genes were found,and an obvious difference in the profiles was found in those treated with high-dose and low-dose tetracycline.The differentially expressed genes of all time points in high-dose group were divided to four clusters,which respectively related to the molecular mechanism of repressed enegy metabolism,enhanced protein synthesis and degradation,impared system of resisting oxidation,signal transduction genes changing to accelerate apoptosis,repressed genes associated of drug metabolism.Conclusion The analysis using toxicological microarray can offer us many clues on molecular level for studying the mechanism of the toxic effect of tetracycline acting on liver.