Changes of collagen and MMP-1 in liver, lung and kidney during growth of mice / 生物工程学报

The high performance liquid chromatography (HPLC) and enzyme-linked immunoassay (ELISA) were used to investigate the changes of collagen and matrix metalloproteinase-1 (MMP-1) in liver, lung and kidney during growth process of mice. The mice from 0 to 18 weeks were used as the research objects. The contents and proportions of hydroxyproline (Hyp), which were used to calculate the collagen contents, in liver, lung and kidney of different weeks were analyzed with HPLC. The contents and activity of MMP-1 in liver, lung and kidney of different weeks were analyzed with ELISA. The results showed that the collagen contents in liver, lung, and kidney were different (Lung(COL)>Kidney(COL)>Liver(COL)), and they all increased first and then decreased with weeks. The collagen contents in liver, lung, and kidney reached the highest level in the ninth (5.52 ng/mg), sixth (54.10 ng/mg) and ninth (19.20 ng/mg) week, respectively. Then it declined slowly from 9 to 18 weeks. The result of ELISA showed that the MMP-1 contents in liver, lung and kidney decreased first and then increased with weeks, and the trend of MMP-1 activity was opposite. It indicated that the increase of collagen contents in the tissues will inhibit the secretion of MMP-1.

DNA methylation and atherosclerosis / 国际脑血管病杂志

Atherosclerosis is a chronic systemic inflammatory disease caused by the interaction of environment and genetic factors. Epigenetic modification is a bridge between environmental factors and genetic factors. DNA methylation is an important regulatory mode of epigenetic modification, which can regulate gene expression at the pre-transcriptional level. Studies have shown that DNA methylation plays an important role in the occurrence and development process of atherosclerosis. Therefore, atherosclerosis-related DNA demethylation, especially carotid atherosclerosis-related DNA demethylation, can be achieved by natural or synthetic DNA demethylases, thereby achieving the purpose of preventing or treating atherosclerotic cerebrovascular disease. This article reviews DNA methylation and its relationship with atherosclerosis and DNA demethylation therapy.

The study on the relationship between the levels of sonic hedgehog and vascular endothelial growth factor in serum and cerebral collateral circulation in patients with symptomatic cerebral artery stenosis / 中国医师进修杂志

Objective To investigate the changes of the levels of sonic hedgehog (SHH) and vascular endothelial growth factor (VEGF) in serum and its relationship with collateral circulation in patients with symptomatic middle cerebral artery stenosis. Methods From January 2015 to January 2018, a total of 268 patients with acute ischemic stroke confirmed as unilateral middle cerebral artery M1 segment (MCA-M1) severe stenosis or occlusion by digital subtract angiography (DSA) were enrolled. The baseline clinical data were collected. According to the establishment of collateral circulation shown by DSA, they were divided into good collateral circulation group (152 patients) and poor collateral circulation group (116 patients). The levels of SHH and VEGF in serum were detected by enzyme linked immunosorbent assay (ELISA), the expression characteristics of SHH and VEGF in serum and the relative factors influencing the establishment of collateral circulation were analyzed. Results The levels of serum SHH and VEGF in good collateral circulation group were significantly higher than those in poor collateral circulation group (P < 0.01). Pearson correlation analysis showed that there was a positive correlation between SHH and VEGF (r=0.758, P < 0.01). Multivariate Logistic regression analysis showed that the levels of serum SHH ( OR=0.310, 95% CI 0.117-0.819, P=0.018) and VEGF ( OR=0.361, 95% CI 0.147-0.887, P=0.026) were independent protective factors for the establishment of collateral circulation. Diabetes ( OR=3.094, 95% CI 1.321-7.245, P=0.009) was independent risk factor for the establishment of collateral circulation. Conclusions The levels of serum SHH and VEGF are closely related to the formation of collateral circulation and they are independent protective factors. SHH may be involved in the establishment of cerebral collateral circulation by regulating the expression of VEGF and diabetes is not conducive to the formation of collateral circulation.

Effects of Tongyuan acupuncture combined with salt and moxibustion Shenque on behavior and hypotha-lamic-pituitary-adrenal axis in postpartum depression rats / 实用医学杂志

Objective To explore the effects of Tongyuan acupuncture combined with herbal medicine and moxibustion on the behavior and hypothalamic-pituitary-adrenal axis-related hormone levels in postpartum de-pression rats.To explore the therapeutic mechanism of this method in the treatment of postpartum depression.Meth-ods A total of 120 SD rats were randomly divided into 6 groups(n=20).The blank group was not treated.The sham operation group was performed without surgery to remove the ovaries. The model group,drug group,routine acupuncture group and experimental group were established in the model of postpartum depression.After modeling successfully,the model group did not interfere with any treatment. The rats in treatment group were treated with fluoxetine hydrochloride. The acupuncture group was treated with acupuncture of Baihui and Taichong. The rats in the experimental group were treated with the method of acupuncture and moxibustion.At the end of the second and fourth week after the initiation of intervention,10 rats were randomly selected from each group.They were sacrificed and the levels of serum corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),cortisol (Cor)were measured.Results The levels of CRH,ACTH and Cor in the model group were higher than those in the blank group(P<0.05).The levels of serum CRH,ACTH and Cor in the drug group,routine acupuncture group and experimental group were lower than those in the model group(P<0.05).The levels of serum ACTH and Cor in the experimental group were lower than those in the drug group and routine acupuncture group(P<0.05).There was no significant difference in serum CRH,ACTH and Cor level between the experimental group and blank group(P >0.05).Conclusions Tongyuan acupuncture combined with salt moxibustion and Shenque have significant effects on the depressive symptoms and serum levels of CRH,ACTH and Cor in postpartum depression rats,which may be re-lated to the regulation of hypothalamic-pituitary-adrenal axis-related hormone levels.

Influence of collagen coating on the biocompatibility of three-dimensional printed implants / 中国组织工程研究

BACKGROUND: Inert biomaterials such as metal usually hold poor biocompatibility and weak bonding force, which is against the effect of dental or bone implants. Therefore, how to improve their biocompatibility has become the research hotspot.OBJECTIVE: To prepare collagen-coated titanium alloy (Ti6Al4V), and to assess its biocompatibility.METHODS: Ti6Al4V served as the matrix, the 3-amino propyl triethoxy silane (KH550) as the crosslinking agent, and the three-dimensional printed titanium alloy coated by collagen type Ⅰ and Ⅱ was prepared,respectively. The coated materials were co-cultured with mouse preosteoblasts MC-3T3-E1 to evaluate its biocompatibility. The effect of different kinds of collagen on the cell differentiation was compared by differential recognition of surface proteins.RESULTS AND CONCLUSION: When the N content on the titanium alloy surface (Φ10 cm) was 8.41%, the cladding quantity of collagen type Ⅰ and Ⅱ was 0.81 and 0.77 mg, respectively. Compared with the bare titanium alloy, the cell adhered well and distributed extensively on the coated titanium alloy,which showed strong viability and fast proliferation.The cells cultured on collagen type Ⅰ coated materials expressed the proteins associated with matrix synthesis, and those on collagen type Ⅱ coated materials expressed the proteins associated with mineralization. These results clarify that the collagen coating can improve the biocompatibility of titanium alloy, and different types of collagens act on different functional proteins.

Collagen quantitation by detection of marker peptides with HPLC-MS / 生物工程学报

A method for quantitation of collagen was established by detecting marker peptide with high performance liquid chromatography-mass spectrometry (HPLC-MS). Theoretical marker peptides were selected by sequence comparison. Bovine collagen type I was digested with trypsin. Marker peptides typical for collagen type I were identified with HPLC-MS. The relationship between the abundance of marker peptides and collagen concentration was established. The results show that GEAGPSGPAGPTGAR and the other 5 peptides showed high resolution during chromatographic separation and high signal intensity during MS analysis. Peptide signal intensity and collagen concentration showed a good linear relationship in the range from 0.1 to 3 mg/mL. Bovine tendon and collagen sponge were used as actual samples and collagen contents were determined as 90.2% and 93.4% respectively. Quantitation of marker peptides of collagen was a feasible method to identify and quantify collagens in medical device research and development.

Purification of recombinant human antithrombin III expressed in a goat mammary bioreactor / 生物工程学报

Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.

Chromatography-assisted refolding of a fusion protein containing multiple disulfide bonds / 生物工程学报

To establish a refolding process for the protein fused with 12-peptide of hirudin and reteplase (HV12p-rPA), we developed an anion-exchange chromatography assisted method to form correct disulfide bonds. After evaluating various parameters by orthogonal experiments with Q Sepharose XL as refolding medium, we found that urea gradient, sample loading size and L-Arg concentration were three major factors to affect the refolding outcomes, and urea gradient was critical to the recovery yield. Meanwhile, enzymatic activity of the refolded protein was decreased by the increase of sample loading size, and the optimal concentration of L-Arg in the eluting buffer was 1 mol/L. Thus, a dual-gradient of urea and pH on the anion-exchange chromatography resulted in remarkable increase of specific fibrinolytic and anti-coagulative activities of the refolded protein. Compared with the dilution method for refolding HV12p-rPA, the present approach was more effective and advantageous.

Investigation on degradation process of collagens during manufacturing procedure of E'jiao / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the degradation process of collagens and identify the key unit operation during manufacturing process of E'jiao.</p><p><b>METHOD</b>Samples in different unit operations were withdrawn, and their amino acid compositions and the molecular weight ranges were determined. The peptide composition was analyzed by high-performance liquid chromatography/mass spectrometry.</p><p><b>RESULT</b>The content of sample during atmospheric condensation unit increased by 16.8% compared to the thermal extraction unit. Gel filtration chromatographic analysis indicated that the degradation process of collagen primarily occurred during the atmospheric condensation unit. The peptides in samples mainly resulted from the degradation of collagens and cytoskeleton proteins such as tubulin, actinin, and so on. The relative abundance of degraded collagens increased with the decrease of no-collagen proteins.</p><p><b>CONCLUSION</b>Collagen degradation mainly occurred during the atmospheric condensation unit, which was the key process affecting the composition of E-jiao.</p>

Changes of Microarousal in OSAHS Patients Before and After Operation / 中国医师杂志

Objective To explore the value of microarousal in the diagnosis and treatment of obstructive sleep apnea-hypopnea syndrome(OSAHS). Methods 120 snorers were monitored by standard polysomnography(PSG). The overnight PSG recordings were also performed 1 year after operation in 24 patients, who had filled in the Epworth Sleep Score(ESS) before and after treatment, to evaluate the improvement of daytime sleepiness. Results After operation, obstructive sleep apnea of the 24 patients disappeared, their ODI 4 decreased from (52?26)/h to (10?8)/h, microarousal decreased from (28?16)/h to (14?6)/h, and the subjective and objective daytime sleepiness was improved. Conclusion The EEG changes of microarousal with at least 3 seconds in OSAHS patients can be recorded by computerized EEG analysis. Microarousal can serve as an index for daytime sleepiness, and be complementary to AHI in the diagnosis and treatment of OSAHS.

Studies on the Bioequialence of Two Preparations of Gemfibrozil / 中国药房

OBJECTIVE:To evaluate the bioequivalence of two preparations of gemfibrozil.METHODS:A single oral dose of gemfibrozil enteric capsule(test preparation)and capsule(reference preparation)was given to20volunteers in an open ran?domized crossover way to study the pharmacokinetics and relative bioavailability.The plasma gemfibrozil concentrations were determined by HPLC method.RESULTS:The pharmacokinetic parameters of test and reference preparations were as follows:T max ,(2.4?0.6)h and(2.3?0.7)h;C max ,(21.8?7.3)?g/ml and(23.7?5.9)?g/ml;T 1/2 ,(2.0?0.4)h and(2.0?0.5)h;AUC 0～12 ,(68.1?13.7)(?g?h)/ml and(68.9?17.4)(?g?h)/ml;AUC 0～∞ (69.7?13.9)(?g?h)/ml and(70.6?17.8)(?g?h)/ml respectively.The relative bioavailability of test preparation was(100.8?15.0)%.The result of statistical analysis on above parameters showed that there was no significant difference between two preparations.CONCLUSION:The two prepa?rations were bioequivalent.

Study on the Bioavailability and Pharmacokinetics of Compound Rifampicin Tablets in Healthy Volunteers / 中国药房

OBJECTIVE:To study the pharmacokinetics and relative bioavailability of compound rifampicin tablets MET_HODS:Plasma levels of rifampicin(RFP),isoniazid(INH)and pyrazinamid(PZA) at different time were determined by HPLC methods,then we drew the time-concentration curves and got the pharmacokinetic parameters and relative bioavailability of test-tablets based on the curves RESULTS:The main pharmacokinetic parameters of RFP,INH and PZA in test-tablets were:Tmax,(1 69?0 60)h,(0 94?0 57)h and(2 36?1 10)h;Cmax,(9 86?2 09)?g/ml,(5 36?1 77)?g/ml and (16 20?4 85)?g/ml;T1/2,(3 43?0 72)h,(2 98?0 75)h and(9 26?1 58)h;AUC0～t,(59 34?13 17)?g/(ml?h),(21 87?14 29)?g/(ml?h) and(212 97?71 52)?g/(ml?h) respectively The main pharmacokinetic parameters of RFP,INH and PZA in control tablets were Tmax,(1 83?0 66)h,(0 86?0 38)h and (2 08?0 97)h;Cmax,(9 98?1 63)?g/ml,(5 60?2 01)?g/ml and (17 79?4 57)?g/ml;T1/2,(3 97?1 58)h,(3 15?0 88)h and (9 36?1 85)h;AUC0～t,(62 46?14 02)?g?h/ml,(21 39?14 53)?g/(ml?h) and (227 09?70 91)?g/(ml?h) respectively The relative bioavailability of test-tablets were (98 47?15 00)%,(103 76?15 80)% and (94 38?12 07)% CONCLUSION:The results of two one-sided tests and rank sum test showed that two formulae were statistically bioequivalent

Assay of Concentration of Isoniazid in Plasma With HPLC / 中国药房

OBJECTIVE:A HPLC method has been developed to determine the concentration of isoniazid in plasma.METH_ODS:The Eclipse XDB-C18 column was used as fix phase and acetonitrile-0.05mol/L ammonium dihydrogen phosphate as mo_bile phase,detection wavelength was 280nm.The plasma sample was injected directly for determination after being deproteinized with 10% trichloroacetic acid and reacted with cinnamaldehyde and abstracted with ether.RESULTS:Good linear relationship was shown from 0.10 to 12.0?g/ml and the averge recovery of isoniazid was 95%～105%.CONCLUSION:The method is rapid,sensitive and is rarely interfered so it can be used in study of pharmacokinetics of isoniazid.