Bone metabolism in patients with osteonecrosis of the femoral head based on etiology and Association Research Circulation Osseous staging / 中国组织工程研究

BACKGROUND:Currently,there is a lack of large sample studies to analyze the bone metabolism level of patients with femoral head necrosis of different etiologies and stages,which is not conducive to the development of better necrosis-promoting repair strategies. OBJECTIVE:To study the bone metabolism of patients with osteonecrosis of the femoral head with different etiologies and Association Research Circulation Osseous(ARCO)stages. METHODS:A retrospective study was performed on 401 patients diagnosed with osteonecrosis of the femoral head as the trial group,and 81 healthy subjects as the control group.The trial group could be divided into three groups according to different etiologies:steroid-induced osteonecrosis of the femoral head,alcoholic osteonecrosis of the femoral head and traumatic osteonecrosis of the femoral head,and were divided into stages Ⅱ/Ⅲ/Ⅳ according to different ARCO stages.Seven bone metabolism-related indicators of all subjects were collected,including bone metabolism-regulating hormone 25-hydroxyvitamin D and bone conversion markers:N-terminal propeptide of type Ⅰ procollagen,degradation product of type Ⅰ collagen,n-terminal middle molecular fragment of osteocalcin,general biochemical markers of bone metabolism:serum calcium,serum phosphorus,serum alkaline phosphatase.The bone metabolism levels of each group were compared and the independent factors associated with osteonecrosis of the femoral head were determined by binary Logistic regression analysis. RESULTS AND CONCLUSION:Compared with the control group,levels of degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen,n-terminal middle molecular fragment of osteocalcin,serum phosphorus and alkaline phosphatase in the trial group were significantly increased(all P＜0.05).Based on the presence or absence of the disease,according to binary Logistic regression analysis,degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen,and n-terminal middle molecular fragment of osteocalcin were independent factors associated with osteonecrosis of the femoral head.The levels of degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen in three groups of patients with different etiologies were higher than normal reference values.The bone metabolism-regulating hormone 25-hydroxyvitamin D and serum calcium in the alcoholic osteonecrosis of the femoral head group were higher than those in the other two groups(P＜0.05).The level of bone metabolism-regulating hormone 25-hydroxyvitamin D in steroid-induced and traumatic osteonecrosis of the femoral head groups was lower than the normal value.There were no significant differences in seven bone metabolism-related indicators in patients with ARCO stages Ⅱ,Ⅲ and Ⅳ osteonecrosis of the femoral head(all P＞0.05),but degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen in these three groups were higher than normal reference values.Bone metabolism-regulating hormone 25-hydroxyvitamin D in patients with ARCO stage Ⅱ and ARCO stage Ⅳ was lower than the normal reference value.It is concluded that the bone metabolism level of osteonecrosis of the femoral head patients was abnormal.The degradation product of type Ⅰ collagen and N-terminal propeptide of type Ⅰ procollagen of osteonecrosis of the femoral head patients with different etiologies and ARCO stages were all higher than the normal reference value,and they were in a state of high bone turnover.Degradation product of type Ⅰ collagen,N-terminal propeptide of type Ⅰ procollagen and n-terminal middle molecular fragment of osteocalcin may be risk factors for the pathogenesis of osteonecrosis of the femoral head.

Punicalagin treats postmenopausal osteoporosis by promoting osteogenesis / 中国组织工程研究

BACKGROUND:Punicalagin has a wide range of effects and high safety,but its effect on osteoblasts and postmenopausal osteoporosis is unknown. OBJECTIVE:To investigate the effect of punicalagin on osteoblasts and postmenopausal osteoporosis. METHODS:The effect of punicalagin on the proliferation of MC3T3-E1 cells was detected.Punicalagin was added to the osteogenic induction medium to detect its effect on osteogenic differentiation.Punicalagin was used to treat ovariectomized rats and Micro CT scan and serum procollagen type 1 N-terminal propeptide test were performed after 3 months to detect the therapeutic effect. RESULTS AND CONCLUSION:Cell counting kit-8 assay showed that punicalagin could promote the proliferation of osteoblasts(P＜0.05).The results of qRT-PCR and western blot showed that punicalagin could promote the mRNA and protein expressions of alkaline phosphatase and Runx2 in osteoblasts(P＜0.05).The results of Micro CT scan and serological test showed that punicalagin could improve bone mineral density,bone volume fraction,trabecular thickness,trabecular number and procollagen type 1 N-terminal propeptide level of ovariectomized rats.To conclude,punicalagin can promote osteoblast proliferation and differentiation,and have therapeutic effects in postmenopausal osteoporosis rats.

Evaluation of Clinical Efficacy of Shengmaisan Granules in Inhibiting Myocardial Fibrosis in Patients with Chronic Heart Failure with Qi-Yin Deficiency Syndrome Based on CMR / 中国实验方剂学杂志

ObjectiveTo evaluate the effect of Shengmaisan granules on myocardial fibrosis in chronic heart failure patients with Qi-Yin deficiency syndrome by cardiac magnetic resonance （CMR） imaging and serological indicators. MethodSixty-six chronic heart failure patients with Qi-Yin deficiency syndrome who visited the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine， Nanjing University of Chinese Medicine from October 2021 to January 2023 were selected. The patients were assigned into a control group （33 cases） and an observation group （33 cases） by the minimization random method. Both groups received standardized Western medicine treatment for heart failure. In addition， the control group was treated with placebo granules， and the observation group with Shengmaisan granules for a course of 6 months. The baseline data， clinical efficacy， TCM symptom scores， serological indicators ［high-sensitivity C-reactive protein （hs-CRP）， soluble growth stimulation expressed gene 2 protein （sST2）， pro-collagen Ⅲ N-terminal peptide （PⅢNP）， interleukin （IL）-6， IL-11， transforming growth factor-β1 （TGF-β1）］， echocardiography ［Left atrial diameter （LAD）， left ventricular end systolic diameter （LVEDs）， left ventricular end diastolic diameter （LVEDd）］ and CMR indicators ［left ventricular ejection fraction （LVEF）， myocardial extracellular volume fraction （ECV）， and longitudinal relaxation time （T1）］ were compared between the two groups. ResultFinally， 31 patients in the control group and 30 patients in the observation group were included. There was no significant difference in baseline data or indicators between the two groups before treatment. Compared with those before treatment， the scores of TCM symptoms （shortness of breath， fatigue， palpitations， spontaneous or night sweats， thirst/dry throat， feverish feeling in palms and soles， and edema in lower limbs）， total score of TCM symptoms， ECV， T1， inflammation/fibrosis indicators （hs-CRP， sST2， PⅢNP， IL-6， IL-11， and TGF-β1） in observation group decreased （P<0.05， P<0.01）， and the scores of TCM symptoms （except feverish feeling in palms and soles）， T1， and inflammation/fibrosis indicators in the control group decreased （P<0.05， P<0.01）. After treatment， the observation group had lower scores of TCM symptoms （except feverish feeling in palms and soles and edema in lower limbs）， ECV， T1， and inflammation/fibrosis indicators than the control group （P<0.05， P<0.01）. After treatment， the total response rate in the observation group was 93.33% （28/30）， which was higher than that （80.65%， 25/31） in the control group （Z=2.976， P<0.01）. There was no significant difference in adverse reactions between the two groups during treatment. ConclusionFor patients with chronic heart failure with Qi-Yin deficiency syndrome， Shengmaisan Granules can alleviate the TCM symptoms， reduce inflammation， and inhibit myocardial fibrosis by regulating the TGF-β1/IL-11 signaling axis.

Value of dual energy CT parameters combined with serum procollagen Ⅰ N-terminal propeptide and beta C-terminal cross-linked telopeptide of type Ⅰ collagen in differential diagnosis of spinal bone metastasis from lung cancer and myeloma / 中国医师进修杂志

Objective:To analyze the value of dual energy CT parameters combined with serum procollagen Ⅰ N-terminal propeptide (PⅠNP) and beta C-terminal cross-linked telopeptide of type Ⅰ collagen (β-CTX) in differential diagnosis of spinal bone metastasis from lung cancer and myeloma.Methods:The clinical data of 54 patients with spinal bone metastasis from lung cancer and 50 patients with myeloma in Jincheng People′s Hospital from October 2019 to March 2021 were analyzed retrospectively. All patients were examined by dual energy CT on the day of admission, and the CT values at the energy levels of 40 to 80 keV (energy interval of 10 keV) were recorded. The serum PⅠNP and β-CTX levels were detected by chemiluminescent assay before treatment. The pathological examination results were taken as gold standard, and the CT values at the energy levels of 40 to 80 keV by dual energy CT and serum PⅠNP and β-CTX levels were compared between 2 groups. Receiver operating characteristic (ROC) curve was used to analyze the differential diagnosis value of the CT values at the energy levels of 40 to 80 keV, serum PⅠNP and β-CTX levels alone and combination.Results:The CT values at the energy levels of 40 to 80 keV by dual energy CT and serum PⅠNP and β-CTX levels in patients with spinal bone metastasis from lung cancer were significantly higher than those in patients with myeloma: 79.86 (61.20, 116.32) HU vs. 58.29 (46.92, 64.03) HU, 64.48 (50.27, 90.08) HU vs. 45.78 (38.59, 56.75) HU, 57.35 (43.31, 78.04) HU vs. 43.62 (36.91, 54.06) HU, 52.05 (42.98, 75.79) HU vs. 41.26 (32.84, 51.76) HU, 45.52 (38.55, 63.59) HU vs. 36.68 (28.72, 49.83) HU, 66.35 (31.15, 81.97) μg/L vs. 31.38 (27.76, 34.50) μg/L and 0.61 (0.48, 0.67) μg/L vs. 0.49 (0.47, 0.52) μg/L, and there were statistical differences ( P<0.05 or <0.01). ROC curve analysis result showed that the sensitivity of the combination of the CT values at the energy levels of 40 to 80 keV by dual energy CT was higher than those alone (83.33% vs. 59.26%, 61.11%, 62.96%, 64.81% and 66.67), the area under the curve (AUC) was also higher than those alone (0.882 vs. 0.798, 0.811, 0.817, 0.801 and 0.773), and there were statistical differences ( P<0.01 or <0.05); the sensitivity of the combination of serum PⅠNP and β-CTX levels was higher than those alone (81.48% vs. 57.41% and 62.96%), the AUC was higher than those alone (0.829 vs. 0.753 and 0.729), and there were statistical differences ( P<0.01 or <0.05); the sensitivity of all indexes combined in the differential diagnosis of spinal bone metastasis from lung cancer and myeloma was higher than those of the combination of the CT values at the energy levels of 40 to 80 keV by dual energy CT, the combination of serum PⅠNP and β-CTX levels (98.15% vs. 83.33% and 81.48%), the same as AUC (0.976 vs. 0.882 and 0.829), and there were statistical differences ( P<0.01); there were no significant differences in the specificity of each index alone and combination ( P>0.05). Conclusions:Compared with myeloma, the CT values at the energy levels of 40 to 80 keV by dual energy CT, serum PⅠNP and β-CTX levels in patients with spinal bone metastasis from lung cancer are increased, and the combination of the above indexes has ideal value in differential diagnosis of the two diseases.

Effectiveness of FibroTouch combined with four hepatic fibrosis biomarkers for evaluation of the liver fibrosis degree among patients with chronic schistosomiasis-induced liver disorders / 中国血吸虫病防治杂志

Objective To evaluate the performance of FibroTouch in combination with four hepatic fibrosis biomarkers for assessment of the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders. Methods A total of 63 patients with chronic schistosomiasis-induced liver diseases admitted to The Third People’s Hospital of Kunshan City from January to March 2021 were enrolled as the observation group, while 50 healthy volunteers receiving health examinations in the hospital during the study period were randomly selected as the control group. The liver stiffness measurement (LSM) was determined using the FibroTouch technique, and the serum levels of four hepatic fibrosis biomarkers were detected using chemilumi-nescence immunoassay, including type IV collagen (IV-C), type III procollagen (PC-III), hyaluronidase (HA) and laminin (LN). The receiver operating characteristic (ROC) curves of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver disorders were plotted and the area under the ROC curve (AUC) was estimated to examine the value of LSM and four hepatic fibrosis biomarkers alone and in combination for assessing the degree of hepatic fibrosis. Results There were 63 subjects in the observation group, including 28 men and 35 women, and the participants had a mean age of (65.34 ± 12.56) years and a mean body mass index (BMI) of (24.47 ± 11.05) kg/m2. There were 50 subjects in the control group, including 22 men and 28 women, and the participants had a mean age of (64.28 ± 13.10) years and a mean BMI of (25.12 ± 11.64) kg/m2. There were no significant differences between the observation and control groups in terms of gender ratio (χ2 = 0.002, P > 0.05), age (t = 0.437, P > 0.05) or BMI (t = 0.303, P > 0.05). The LSM [(8.65 ± 5.22) vs. (3.24 ± 1.10) kPa; t = 8.013, P < 0.05], IV-C [(51.80 ± 9.45) vs. (30.10 ± 10.34) ng/L; t = 11.506, P < 0.05], PC-III [(77.28 ± 17.22) vs. (48.62 ± 9.54) ng/L; t = 11.224, P < 0.05], HA [(39.55 ± 5.32) vs. (84.89 ± 10.34) ng/L; t = 30.158, P < 0.05] and LN [(99.47 ± 7.37) vs. (61.93 ± 9.80) ng/L; t = 22.496, P < 0.05] were significantly greater in the observation group than in the control group, and Spearman correlation analysis showed that the degree of liver fibrosis positively correlated with LSM (rs = 0.675, P < 0.01), IV-C (rs = 0.421, P < 0.01), PC-III (rs = 0.517, P < 0.01), HA (rs = 0.550, P < 0.01) and LN (rs = 0.539, P < 0.01) among patients with chronic schistosomiasis-induced liver diseases. ROC curve analysis revealed that the AUC of LSM for assessment of the hepatic fibrosis degree was 0.884 (P < 0.001), and the LSM cutoff, sensitivity and specificity were 11.75 kPa, 71.43% and 84.00% at the highest Youden index, respectively. In addition, the AUC of four hepatic fibrosis biomarkers for assessment of the hepatic fibrosis degree was 0.577 to 0.670, with 70.174 to 115.237 ng/L cutoff values, 17.46% to 68.25% sensitivity and 71.01% to 96.00% specificity. In addition, the sensitivity and specificity of LSM combined with four hepatic fibrosis biomarkers were 92.06% and 95.07% for assessment of the hepatic fibrosis degree among patients with chronic schistosomiasis-induced liver diseases. Conclusion FibroTouch in combination with detection of four hepatic fibrosis biomarkers has a high sensitivity and specificity for assessing the degree of hepatic fibrosis among patients with chronic schistosomiasis-induced liver diseases, which deserves widespread clinical uses.

Changes of fibrinogen and collagen metabolism after cardiac surgery and their relationship with postoperative atrial fibrillation: A prospective cohort study / 中国胸心血管外科临床杂志

@#Objective    To investigate the changes of fibrinogen and classical markers of collagen metabolism [carboxy-terminal propeptide of type Ⅰ procollagen (PICP) and carboxy-terminal cross-linked peptide of type Ⅰ collagen (ICTP)] in peripheral blood and pericardial drainage after coronary artery bypass grafting (CABG) and/or heart valve replacement (VR), and to evaluate their relationship with postoperative atrial fibrillation (POAF) after cardiac surgery. Methods    Patients who underwent CABG and/or VR in the Heart Center of Beijing Chao-Yang Hospital from March to June 2021 were included. Peripheral blood and pericardial drainage fluid samples were collected before surgery and at 0 h, 6 h, 24 h and 48 h after surgery to detect PICP, ICTP and fibrinogen levels, and preoperative, intraoperative and postoperative confounding factors were also collected. PICP, ICTP and fibrinogen levels were measured by enzyme-linked immunosorbent assay (ELISA). Results    A total of 26 patients with 125 blood samples and 78 drainage samples were collected. There were 18 males and 8 females with an average age of 64.04±7.27 years. The incidence rate of POAF was 34.6%. Among the factors, the fibrinogen level in pericardial drainage showed two peaks within 48 h after operation (0 h and 24 h after operation) in the POAF group, while it showed a continuous downward trend in the sinus rhythm (SR) group, and the change trend of fibrinogen in pericardial drainage was significantly different over time between the two groups (P=0.022). Fibrinogen in blood, PICP and ICTP in blood and drainage showed an overall decreasing trend, and their trends over time were not significantly different between the two groups of patients (P>0.05). Univariate analysis showed that fibrinogen at 24 h and 48 h after pericardial drainage, fibrinogen in preoperative blood, PICP immediately after surgery and right atrial long axis diameter were significantly higher or longer in the POAF group than those in the SR group. Multiple regression showed that fibrinogen≥11.47 ng/mL in pericardial drainage 24 h after surgery (OR=14.911, 95%CI 1.371-162.122, P=0.026), right atrial long axis diameter≥46 mm (OR=10.801, 95%CI 1.011-115.391, P=0.049) were independent predictors of POAF. Conclusion    This study finds the regularity of changes in fibrinogen and collagen metabolic markers after CABG and/or VR surgery, and to find that fibrinogen in pericardial drainage 24 h after surgery is a potential novel and predictive factor for POAF. The results provide a new idea for exploring the mechanism of POAF, and provide a research basis for the accurate prediction and prevention of clinical POAF.

Development and evaluation of a chemiluminescence immunoassay for type Ⅰ procollagen N-terminal peptide / 生物工程学报

To develop a magnetic nanoparticle chemiluminescence immunoassay (CLIA) for the determination of type Ⅰ procollagen N-terminal peptide (PINP) in human serum, we expressed a recombinant PINP-α1 protein in Corynebacterium glutamicum and used it as an immunogen to immunize BALB/c mice. We obtained three hybridoma cell lines that stably secret antibody against PINP-α1 protein. After further pairing and screening, we chose a monoclonal antibody 8C12 coupled with biotin as the capture antibody, and a monoclonal antibody 1F11 labeled horseradish peroxidase as the detection antibody. The antibodies combined with the serum samples, forming a sandwich complex which was used to detect the concentration of PINP in serum. After optimizing the conditions, we determined that the best working concentration of the capture antibody and the detection antibody were 3 μg/mL, and the incubation time was 30 minutes. The quantitative assay had a detection range of 5-1 100 ng/mL, with recovery rates between 93%-107% and the minimum detection limit of 1.22 ng/mL achieved. The intra-and inter-assay precisions were lower than 10%. The correlation coefficient of PINP results between this CLIA method and the Roche electrochemiluminescence immunoassay system was 0.906 2. Therefore, this CLIA method is specific and can be used to quantitatively detect the content of PINP in serum, which has the potential to become an auxiliary approach for bone disease examination.

Lentinula edodes extract inhibits matrix metalloproteinase expression and increases typeⅠprocollagen expression via the p38 MAPK/c-Fos signaling pathway in ultraviolet A and B-irradiated HaCaT keratinocytes

Objective:To determine the effect of Lentinula edodes extract on ultraviolet (UV) A and UVB-induced changes in matrix metalloproteinase (MMP) and typeⅠprocollagen expression using human immortalized HaCaT keratinocytes. Methods:Lentinula edodes ethanol extract (LEE) was obtained by extraction with 80％ ethanol for 4 h at 80 ℃. Effect of LEE on UV-induced alteration on the expression and production of MMPs and typeⅠprocollagen in keratinocytes was investigated using ELISA, RT-PCR, and Western blotting assay. To determine the underlying mechanism of LEE-mediated effects, mitogen-activated protein kinase (MAPK) and activator protein 1 signaling pathways were analysed by Western blotting assay. Results:LEE significantly inhibited the expression of MMP-1 and MMP-9 and increased the expression of typeⅠprocollagen in UVA and UVB-irradiated HaCaT keratinocytes. The phosphorylation levels of p38 were significantly inhibited by LEE whereas it did not affect c-Jun N-terminal kinase and extracellular signal-regulated kinase phosphorylation. Suppression of p38 phosphorylation was also accompanied by downregulation of UVA and UVB-induced increase in c-Fos. Conclusions:LEE effectively inhibits the expression of MMP-1 and MMP-9 and increases typeⅠprocollagen production through the p38 MAPK/c-Fos signaling pathway in UVA and UVB-irradiated HaCaT keratinocytes. This findings suggest that Lentinula edodes may be developed as a cosmetic material to suppress UV exposure-mediated skin aging.

Study of bone turnover markers, procollagen type 1 N-terminal propeptide and osteocalcin in healthy and type 2 diabetes mellitus postmenopausal women

Background: Individuals with diabetes mellitus are at increased risk of metabolic bone disease due to decrease in bone strength and quality. Several bone turnover markers like serum procollagen type I N propeptide (P1NP) and serum osteocalcin are powerful tools for studying osteoporosis and fracture risk across population to provide diagnostic and prognostic information of bone health. The aim of this study was to recognize possible correlation of levels of serum P1NP and osteocalcin in type-2 diabetic (T2DM) postmenopausal women as compared to healthy postmenopausal women.Methods: The study included 100 proven cases of type-2 diabetic postmenopausal women with age matched healthy postmenopausal women as controls. P1NP, osteocalcin, and other relevant parameters were measured. Differences between diabetics and controls were analyzed.Results: The body mass index was higher in diabetic group as compared to controls. The HbA1c% was (6.94±1.43) in diabetic group and (5.57±1.21) in non-diabetics. Low serum level of 25 (OH) D was observed both in diabetic and non-diabetic groups but significantly lower in T2DM. Procollagen type 1 N propeptide was lower in diabetic group (37.59±17.20 ng/mL) as compared to non-diabetic (52.14±24.82 ng/mL). Osteocalcin was lower (15.64±8.06 ng/ml) as compared to non-diabetic group (21.85±9.12 ng/ml). Lower osteocalcin and P1NP levels found in this study suggests slower bone metabolism with reduced bone formation in postmenopausal diabetics.Conclusions: Serum procollagen type 1 N propeptide and osteocalcin in postmenopausal diabetic women were lower as compared to non-diabetic group.

Bone Turnover Markers and Sphingosine-1-phosphate levels among the Chinese Community in Selangor, Malaysia and its Correlation with Bone Density

@#Introduction: Prevention of osteoporotic fracture requires identification of individuals at high risk. Bone mineral density(BMD) is commonly used to estimate fracture probability despite inadequate predictive discrimination ability. Sphingosine-1-phosphate(S1P), a new marker of bone metabolism and bone turnover markers(BTM) such as procollagen-type-1 amino-terminal propeptide(P1NP) and C-terminal telopeptide of type I collagen(CTX) may complement current assessment. The study determined P1NP, CTX and S1P levels and their correlation with BMD, 25-hydroxyvitamin D (25(OH)D) and parathyroid hormone(PTH) in selected subjects. Method: A cross-sectional study involving Malaysian Chinese men and women aged 50-90 years old from Puchong and Kajang, Selangor. Each subject had BMD determined by dual-energy x-ray absorptiometry and blood samples taken for 25(OH)D, PTH, P1NP, CTX and S1P. Results: A total of 131 subjects [45(34.4%) males and 86(65.6%) post-menopausal women] with median age of 65(IQR=17) were recruited. P1NP and CTX were significantly higher in post-menopausal women (P1NP=61.71 ng/ml, CTX=0.489 ng/ml) compared to men (P1NP=46.94 ng/ml, CTX=0.381 ng/ml). P1NP and CTX differed significantly according to BMD categories with values highest in osteoporosis. S1P between men (2.12±0.75 µmol/L) and post-menopausal women (1.96±0.68 µmol/L) did not differ significantly and did not differ according to BMD categories. S1P did not correlate with BMD, P1NP, CTX and 25(OH)D. P1NP and CTX negatively correlated with BMD at all measured sites but not 25(OH)D. Conclusion: CTX and P1NP, but not S1P negatively correlated with BMD. CTX and P1NP were highest in those with osteoporosis. In this group of Malaysian Chinese subjects, CTX and P1NP rather than S1P reflects bone health.

Association between col1a2 polymorphism and the occurrence of pelvic organ prolapse in Brazilian women / Associação entre polimorfismo do gene col1a2 e a ocorrência do prolapso de órgãos pélvicos em mulheres brasileiras

Abstract Objective To evaluate the rs42524 polymorphism of the procollagen type I alpha (α) 2 (COL1A2) gene as a factor related to the development of pelvic organ prolapse (POP) in Brazilian women. Methods The present study involved 112 women with POP stages III and IV (case group) and 180 women with POP stages zero and I (control group). Other clinical data were obtained by interviewing the patients about their medical history, and blood was also collected from the volunteers for the extraction of genomic DNA. The promoter region of the COL1A2 gene containing the rs42524 polymorphism was amplified, and the discrimination between the G and C variants was performed by digestion of the polymerase chain reaction (PCR) products with the MspA1I enzyme followed by agarose gel electrophoresis analysis. Results A total of 292 women were analyzed. In the case group, 71 had the G/G genotype, 33 had the G/C genotype, and 7 had the C/C genotype. In turn, the ratio in the control group was 117 G/G, 51 G/C, and 11 C/C. There were no significant differences between the groups. Conclusion Our data did not show an association between the COL1A2 polymorphism and the occurrence of POP.

Resumo Objetivo Avaliar o polimorfismo rs42524 do gene pró-colágeno tipo I alfa (α) 2 (COL1A2) como fator relacionado ao desenvolvimento de prolapso de órgãos pélvicos (POP) em mulheres brasileiras. Métodos O estudo envolveu 112 mulheres com POP nos estádios III e IV (grupo caso) e 180 mulheres com POP nos estádios zero e I (grupo controle). Outros dados clínicos foramobtidos pormeio de entrevistas comas pacientes sobre seu históricomédico, e o sangue das voluntárias também foi coletado para extração de DNA genômico. A região promotora do gene COL1A2 contendo o polimorfismo rs42524 foi amplificada, e a discriminação entre as variantes G e C foi realizada por digestão dos produtos de reação em cadeia da polimerase (RCP) com a enzima MspA1I, seguida de análise por eletroforese em gel de agarose. Resultados Foram analisadas 292 mulheres. No grupo caso, 71 tinham o genótipo G/G, 33 tinham o genótipo G/C, e 7 tinham o genótipo C/C. Por sua vez, a relação no grupo controle foi de 117 G/G, 51 G/C e 11 C/C. Não houve diferenças significativas entre os grupos. Conclusão Nossos dados não mostraram associação do polimorfismo do gene COL1A2 com a ocorrência de POP.

Expression of PLOD2 in esophageal squamous cell carcinoma and its correlation with invasion and metastasis / 中华病理学杂志

Objective@#To investigate PLOD2 expression in esophageal squamous cell carcinoma, and to explore the potential mechanism by which PLOD2 promotes tumor metastasis.@*Methods@#The expression of PLOD2 in 60 cases of esophageal squamous cell carcinoma (the patients were collected at the first Affiliated Hospital of Xinxiang Medical University, from January 2016 to December 2017) was investigated by immunohistochemistry. Fibrillar collagen formation and collagen deposition were detected by picrosirius red staining. Correlation of PLOD2 expression with clinical pathologic features of the patients was performed using χ2 test and Kaplan-Meier analysis. After EC-109 cells were transfected with LV-vector and LV-over/PLOD2, the expression of PLOD2 was detected by real time PCR and the impact of POLD2 on invasion in EC-109 cells was determined by transwell migration and invasion assays. The expression of PLOD2/AKT epithelial-to-mesenchymal transition signal pathway related proteins was detected by Western blot.@*Results@#The expression level of PLOD2 in esophageal squamous cell carcinoma was 81.7% (49/60 cases),higher than their paired noncancerous tissues(8.3%, 5/60; P<0.01), and correlated significantly with tumor depth of invasion and nodal metastasis (P<0.01). Picrosirius red staining showed that collagen deposition was increased and the degree of fibrillar organization was enhanced in carcinoma tissues that had higher PLOD2 expression. Transwell migration and invasion assays showed that PLOD2 significantly promoted the migration and invasion ability of EC-109 cells. Western blot showed that PLOD2 significantly increased the expression levels of p-FAK, p-AKT and vimentin in EC-109 cells.@*Conclusions@#Esophageal squamous cell carcinoma has a high expression of PLOD2 that correlates with tumor invasion and lymph node metastasis. PLOD2 promotes invasion and metastasis of esophageal squamous cell carcinoma through epithelial-to-mesenchymal transition via FAK/AKT signal pathway.

The role and research progress of PLOD2 in tumor metastasis / 医学研究生学报

Tumor invasion and metastasis are the main factors leading to poor prognosis of cancer patients.In recent years, studies have found that collagen fiber remodeling in extracellular matrix(ECM) can promote tumor invasion and metastasis . Lysine hydroxylase 2(PLOD2) is the only lysine hydroxylase that changes the pattern of collagen fiber cross-linking and remodeling. It is highly expressed in many tumors and is closely related to tumor invasion and metastasis,Therefore, in-depth study of the role and mechanism of PLOD2 in tumor metastasis is of great scientific value for judging tumor prognosis and preventing tumor invasion and metastasis.This article reviews the recent advances in the role and potential mechanisms of PLOD2 in tumor invasion and metastasis.

Age-related Reference Intervals for Total Collagen-I-N-terminal Propeptide in Healthy Korean Population

BACKGROUND: Procollagen type I N-terminal propeptide (PINP) is one of the most clinically useful bone formation biomarkers. Therefore, the purpose of this study was to independently evaluate the performance of automated total PINP assay and established age- and gender-specific reference intervals for PINP in healthy Korean population. METHODS: The imprecision, linearity, and detection capability of Elecsys total PINP assay was determined and reference interval was established using 599 serums from Korean population with normal bone mineral densities based on bone densitometry. Age groups were divided into 20s, 30s, 40s, 50s, 60s and over. RESULTS: Elecsys total PINP had excellent performance in imprecision, linearity, and detection capability. When partitioning age groups in Korean male and female populations, there was significant difference in total PINP between different age groups. In male populations, PINP level was decreased with increasing age, then it remained steady after middle-age. In female populations, there was a decreasing tendency similar to that in the male population with a sharp increase in the 50 to 59 age group. CONCLUSIONS: Elecsys total PINP assay showed precise and reliable performance in our study. We established age-related PINP reference intervals for Korean male and female population with normal bone mineral densities.

Correlation between serum LN, Ⅳ C, P Ⅲ NP, HA and the severity as well as the prognosis of idiopathic pulmonary fibrosis / 中华急诊医学杂志

Objective To investigate the correlation between serum laminin (LN),type collagen (ⅣC),type procollagen N-terminal peptide (PⅢNP),hyaluronic acid (HA) and the severity as well as the prognosis of idiopathic pulmonary fibrosis (IPF).Methods From February 2015 to July 2016,160 IPF patients and 160 healthy subjects as controls were enrolled in this retrospective study.Serum LN,ⅣC,PⅢNP,and HA were analyzed in IPF patients and healthy controls.Pulmonary function test and chest high resolution computed tomography (HRCT) were carried out in IPF patients.Demographics and clinical characteristics,the percentage of forced vital capacity in the prediction value (FVC％pred),the percentage of diffusing capacity of the lung for carbon monoxide in the prediction value (DLCO％pred),and HRCTscore were collected.IPF patients were followed up.Results (1)There were no significant difference between two groups in age and sex ratio.The proportion of smoker in IPF patients was significantly higher than that in healthy controls(P ＜ 0.01).(2)Serum LN(P ＜ 0.01),ⅣC(P ＜ 0.01),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) were significantly increased in the patients with IPF compared with the healthy controls.(3)Serum LN,ⅣC,PⅢNP,and HA of IPF patients positively correlated with HRCT score,all P ＜ 0.01,and negatively correlated with FVC％pred and DLCO％pred (all P ＜ 0.05).(4)Serum LN(P ＜ 0.01),ⅣC(P ＜ 0.05),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) in acute exacerbation IPF patients were significantly higher than those in the stable IPF patients.(5)Serum LN(P ＜ 0.01),ⅣC(P ＜0.01),PⅢNP(P ＜ 0.01),and HA(P ＜ 0.01) in the survived patients were significantly lower than those inthe dead patients.Conclusions Serum LN,ⅣC,PⅢNP,and HA may reflect IPF prognosis and the severity of IPF.

Protective effects of Rheum officinale Ball. as a main agent in a traditional Chinese medicine recipe com-bined with sequential hemoperfusion in patients with acute paraquat poisoning / 实用医学杂志

Objective To investigate the clinical value of Rheum officinale Ball.as a main agent in a tra-ditional Chinese medicine recipe combined with continuous hemoperfusion in patients with acute paraquat poison-ing(APP). Methods A total of 124 patients with APP were divided into three groups:A group(40 cases),B group(42 cases)and C group(42 cases). Before,3 days and 7 days after treatment,PⅢNP,CⅣ,TGF-β1, MDA,SOD and SOFA scores were observed. The correlation between PⅢNP and the concentration of paraquat were analyzed,and the outcomes in 28 days were compared.Results In admission,there were no differences in PⅢNP,CⅣ,TGF-β1,MDA,SOD and SOFA scores among three groups(P>0.05).After treatment for 3 days, MDA and SOFA scores were elevated in three groups compared with these three scores in the three groups at admis-sion. Group A was increased significantly;the activity of SOD were decreased,especially in A group decreased most significantly(P<0.05);After 7 days'treatment,MDA and SOFA scores were lower than those of 3 days af-ter treatment in three groups,especially in C group decreased most significantly(P < 0.05),the activity of SOD were higher than 3 days after treatment,especially in C group increased most significantly(P<0.05);After 3,7 days treatment,PⅢNP,CⅣ,TGF-β1 were increased gradually,C group increased slowly(P<0.05).PⅢNP in patients with APP was positively correlated with the concentration of paraquat(P < 0.05). The mortality rate of three groups were 65.00%,45.24%and 23.81%,there were significant difference among three groups(P<0.05). Conclusions Rheum officinale Ball.as a main agent in a traditional Chinese medicine recipe combined with con-tinuous hemoperfusion in patients with APP could decrease PⅢNP,CⅣ,TGF-β1 and MDA. The can enhance the expression of SOD and reduce the mortality rate of patients.

Oligomeric Procyanidins (OPCs) Inhibit Procollagen Type I Secretion of Fibroblasts

Wound healing is composed of a complex process that requires harmonies of various cell populations where fibroblasts play the main role. Oligomeric procyanidins (OPC) are main components of grape (Vitis vinifera) seed extracts, and recent studies showed OPC's effects on inflammation, cell migration, and proliferation. We investigated the effect of OPC on fibroblasts to regulate wound healing process. Human dermal fibroblast known as Hs27 cells were treated with various concentrations of OPC (0, 2.5, 5, 10, and 20 µg/µl). Cell cytotoxicity was evaluated by the Cell Counting Kit assay, and the expression levels of secreted procollagen were analyzed. Procollagen levels in OPC treated cells exposed to transforming growth factor beta 1 (TGF-β1) or ascorbic acid were evaluated using Western blot and immunocytochemistry. Relative mRNA expressions of procollagen, molecular chaperone such as HSP47, P4H were determined by real-time PCR in OPC treated cells. OPC showed no cytotoxicity on Hs27 cells at every concentration but inhibited procollagen secretion in a dose-dependent manner. The inhibitory effect also appeared under TGF-β1 induced collagen overproduction. Immunocytochemistry showed that higher levels of intracytoplasmic procollagen were accumulated in TGF-β1 treatment group, whereas ascorbic acid induced a release of accumulated procollagen under OPC treatment. The mRNA expressions of procollagen, molecular chaperone were not affected by OPC, but procollagen level was increased when exposed to TGF-β1. OPC inhibits procollagen secretion from fibroblasts with no effects on cell proliferations even under the environment of TGF-b1-induced collagen overproduction. OPC could regulate the diseases and symptoms of abnormal overabundant collagen production.

Effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma on immune hepatic fibrosis, proliferation of HSC-T6 and expression of α-SMA and Procollagen I / 中国中药杂志

To compare the effects of Curcumae Rhizoma/vinegar-processed Curcumae Rhizomaon immune hepatic fibrosis, proliferation of HSC-T6, and expressions of α-SMA and Procollagen I. The immunological liver fibrosis model was prepared through intraperitoneal injection with porcine serum 0.5 mL in each rat, twice a week, for 14 weeks. Expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN, HA and HYP, MDA in liver tissues were observed after administration of Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma (0.95, 1.90 g•kg⁻¹). The pathological changes in liver tissues were observed by HE staining. Masson staining and Sirius red staining were used to observe the expression of collagen in rat liver. HSC-T6 was cultured, and the proliferation of HSC-T6 was determined by MTT assay at different concentrations in 12, 24, 36, 48 h. The expressions of α-SMA and Procollagen I were detected by Real-time PCR. The results showed that expressions of serum ALT, AST, PC-Ⅲ, IV-C, LN and HA in Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma groups (0.95, 1.90 g•kg⁻¹) were significantly lower than model group; in terms of effect, vinegar-processed Curcumae Rhizoma group was superior to Curcumae Rhizoma group. Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma containing serum could inhibit the proliferation of HSC-T6 in a dose-effect and time-effect manner. Expressions of α-SMA and Procollagen I in HSC-T6 were decreased after 24 h, especially in 20% vinegar-processed Curcumae Rhizoma containing serum group (P<0.01). Both Curcumae Rhizoma/vinegar-processed Curcumae Rhizoma could reduce immune hepatic fibrosis to varying extent. Their anti-hepatic fibrosis mechanism may be correlated with inhibition of the proliferation of HSC-T6, and reduction of the formation of extracellular matrix and promotion of its degradation.

Wound healing activity of Ullucus tuberosus, an Andean tuber crop

Objective: This study was designed to investigate the wound healing activity of aqueous extracts of Ullucus tuberosus (U. tuberosus) using in vitro models. Methods: Lyophilized pulp and acetone extracts of U. tuberosus were produced using ultrasound extraction. The capacity for collagenase activation was evaluated using fluo-rescence detection of the enzymatic activity. Then, the influence of U. tuberosus extracts on cell proliferation, cell migration and synthesis of the extracellular matrix (ECM) proteins, metalloproteinase (MMP-1) and pro-collagen was analyzed using human dermal fibroblasts in culture. Results: An increase in collagenase activity of 12%supports the utility of U. tuberosus as an agent for scar treatment. In addition, the extracts showed an increase in the pro-liferation and migration of human dermal fibroblasts and the production of pro-collagen and MMP-1 after treatment with U. tuberosus extracts. The increase in proliferation, migration and pro-collagen levels positively influenced the regeneration of scarless tissue during the proliferation phase, whereas the increase in MMP-1 may have favored the wound healing process during the remodeling and cellular differentiation phases. Conclusion: The results of this study show for first time that U. tuberosus is a promising candidate to support scarless tissue regeneration.

Clinical value of serum N-MID,total TP1NP,β-CTx detection combination with whole-body bone scintigraphy in early diagnosis of bone metastasis in patients with malignant tumor / 国际检验医学杂志

Objective To investigate the clinical value of bone metabolism biochemical marker N-MID,TP1NP and beta-CTx combined with whole body bone scintigraphy in early diagnosis of bone metastasis of tumor.Methods The concentration of the 3 markers were measured by the electrochemical luminescence analysis method in 30 cases of healthy control group and 210 cases of patients with malignant tumor,which were divided into non bone metastasis group(45 cases) and bone metastasis group(165 cases).The bone metastasis group were divided into 4 grades(0-grade Ⅲ) by Soloway classification according to whole body bone imaging.Results The levels of serum N-MID,TP1NP and beta-CTx in 165 malignant tumor patients with bone metastasis were significantly higher than in 45 malignant tumor patients with bone metastasis and in 30 healthy control group,the difference was statistically significant(P<0.05).With the increase of the number of metastatic lesions in the bone metastasis group,the serum levels of N-MID,TP1NP,and beta-CTx were increased gradually,and they were positively correlated with the progression of the disease.According to the analysis of ROC curve,the cut-off value,sensitivity and specificity in the diagnosis of tumor bone metastasis were 17.59 ng/mL,70.3％,88.9％ for serum N-MID,43.04 ng/mL,78.2％,95.6％ for TP1NP,and 0.48 ng/mL,73.9％,93.3％ for beta-CTx.Under the ROC curve(AUC) was 0.831 for serum N-MID,0.890 for TP1NP,and 0.869 for beta-CTx.The sensitivity and specificity of three bone metabolic markers in the diagnosis of bone metastasis of malignant tumor were significantly higher.Conclusion Bone metabolism biochemical markers:Serum N-MID,TP1NP and beta-CTx for diagnosis of bone metastasis of malignant tumor are sensitive,accurate and simple,which can significantly improve the efficiency of diagnosis of bone metastasis,and can be combined with whole-body bone scintigraphy in early diagnosis of bone metastasis with malignant tumor.