Myricetin attenuates renal fibrosis by activating Nrf2/HO-1 pathway to inhibit oxidative stress / 药学学报

This paper investigates the effect of myricetin (MYR) on renal fibrosis induced by unilateral ureteral obstruction (UUO) and common bile duct ligation (CBDL) in mice and its mechanism. The animal experiment has been approved by the Ethics Committee of China Pharmaceutical University (NO: 2022-10-020). Thirty-five ICR mice were divided into control, UUO, UUO+MYR, CBDL and CBDL+MYR groups. H&E and Masson staining were used to detect pathological changes in kidney tissues. Western blot (WB) was used to detect the expression of fibrosis-related proteins in renal tissue, and total superoxide dismutase (SOD) activity detection kit (WST-8) was used to detect the changes of total SOD in renal tissue of CBDL mice. In vitro, HK-2 cells and transforming growth factor beta 1 (TGF-β1, 10 ng·mL-1) were used to induce fibrotic model, and high glucose (30 mmol·L-1) was used to induce oxidative stress model, and then treated with different concentrations of MYR, WB was used to detect the expression of fibrosis and oxidative stress-related proteins, while NIH/3T3 cells were treated with different concentrations of MYR, and their effects on cell proliferation were detected by 5-bromo-2′-deoxyuridine (Brdu). The results showed that the renal lesions in UUO group and CBDL group were severe, collagen deposition was obvious, the expression of collagen-Ⅰ (COL-Ⅰ), α-smooth muscle actin (α-SMA), fibronectin (FN), vimentin and plasminogen activator inhibitor-1 (PAI-1) protein was up-regulated, and the activity of SOD enzyme in CBDL group was significantly decreased. MYR partly reversed the above changes after treatment. MYR inhibited the proliferation of NIH/3T3 cells but had no effect on the proliferation of HK-2 cells, and decreased the upregulation of PAI-1, FN and vimentin in HK-2 cells stimulated by TGF-β1. MYR can also up-regulate the down-regulation of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in HK-2 cells stimulated by high glucose. To sum up, MYR can improve renal fibrosis in vivo and in vitro, probably by inhibiting the proliferation of fibroblasts and activating Nrf2/HO-1 signal pathway to inhibit oxidative stress.

A prospective study of the perforator evaluation and eccentric design of anterolateral thigh flap based on superficial fascial perforators assisted by modified computed tomography angiography / 中华烧伤杂志

Objective: To explore the preoperative whole perforator evaluation and intraoperative eccentric design of anterolateral thigh flap (ALTF) based on superficial fascial perforators by modified computed tomography angiography (CTA), and the clinical effects were observed. Methods: A prospective observational study was adopted. Twelve patients with oral and maxillofacial tumors and 10 patients with open injury of the upper limb with large soft tissue defects were hospitalized in the Department of Hand & Microsurgery and Department of Oral & Maxillofacial Surgery of Affiliated Hospital of Binzhou Medical University from January 2021 to July 2022, with 12 males and 10 females, aged from 33 to 75 years, an average age of 56.6 years. The wounds of the patients with oral and maxillofacial tumors were reconstructed by ALTF after the extensive tumor resection and radical cervical lymph node dissection in the same stage; the wounds of the patients with skin and soft tissue defect on the upper limb were covered by ALTF in stage Ⅱ after debridement in stage Ⅰ. After debridement, the area of wound was 3.5 cm×3.5 cm-25.0 cm×10.0 cm and the area of the required flap area was 4.0 cm×4.0 cm-23.0 cm×13.0 cm. Modified CTA scan was performed on the donor site of ALTF before the operation, with the parameters of modified CTA being set to mainly reduce the tube voltage and tube current, and increase the contrast dose and the dual phase scan. The acquired image data were sent to GE AW 4.7 workstation and adopted the volume reconstruction function for visual reconstruction and evaluation of the whole perforator. The information of perforator and source artery was marked on the body surface before operation according to the above evaluation. During the operation, an eccentric flap centered on the visible superficial fascia whole perforator was designed and cut according to the desired flap area and shape. The donor sites of the flap were repaired by direct sutures or full-thickness skin grafts. The total radiation dose was compared between the modified CTA scan and the traditional CTA scan. The distribution of outlet point of perforator of double thighs, the length and direction of superficial fascia perforators based modified CTA were recorded. The type, number, and origin of the target perforator, distribution of of outlet point of perforator, and the diameter, course, and branch of the source artery observed before the operation were compared with those observed during the operation. The healing of donor site wound and the survival of flaps in recipient site were observed after operation. The texture and appearance of flap, oral and upper limb functions, and the functions of femoral donor sites were followed up. Results: The total radiation dose of modified CTA scan was lower than that of the traditional CTA scan. A total of 48 perforators of double thighs were observed, among which, 31 (64.6%) perforators went outward and downward, 9 (18.8%) perforators went inward and downward, 6 (12.5%) perforators went outward and upward, and 2 (4.2%) perforators went inward and upward, and the average length of superficial fascia perforators was 19.94 mm. The preoperative observed type, number, and source of the perforator, the distribution of the outlet point of the perforator, diameter, course, and branches of the source artery were basically consistent with the intraoperative exploration. The types of 15 septocutaneous (including musculoseptocutaneous) perforators and 10 musculocutaneous perforators observed before the operation was consistent with intraoperative exploration. The distance between the mark of the surface perforator point and the actual exit point of the perforator during operation was (0.38±0.11) mm. All flaps survived without vascular crisis. The donor site wounds of 5 cases of skin grafting and 17 cases of direct suturing wounds healed well. The postoperative follow-up was 2 months to 1 year, with an average of 8.2 months, the flaps were soft and slightly bloated; the function of diet and mouth closing was accessible in patients with oral and maxillofacial tumors, the speech function was mildly impaired in patients with tongue cancer, but they could complete basic oral communication; the wrist and elbow joints and forearm rotation function were not significantly limited in patients with upper limb soft tissue injuries; there was no obvious tightness in the donor sites, and the function of the hip and knee joints was not limited. Conclusions: The whole perforator and even the subcutaneous perforator of the donor site of ALTF can be evaluated by modified CTA, and the flap can be used in oral or maxillofacial reconstruction and repair of skin and soft tissue defects of upper limbs to achieve good results. By clarifying the type, number, and source of the perforator, the distribution of the outlet point of the perforator, diameter, course, and branches of the source artery before the operation, the eccentric design of the ALTF based on the superficial fascia perforator was realized. This study has strong guiding value.

Intraoperative assisted O-arm navigation imaging for unstable pelvic fractures in INFIX / 中国骨伤

OBJECTIVE@#To investigate the effect of internal external fixator assisted O-arm navigation imaging in the treatment of unstable pelvic fractures.@*METHODS@#From May 2019 to November 2019, 15 patients with unstable pelvic fractures were treated by intraoperative O-arm navigation imaging using INFIX technology. There were 6 males and 9 females. The age ranged from 24 to 66 years old. The course of disease ranged from 2 to 14 days. According to Tile classification, there were 1 case of B1 type, 8 cases of B2 type, 3 cases of C1 type, and 3 cases of C2 type. According to Young-Burgess classification, there were 8 cases of LC, 1 case of APC, 4 cases of VS, 2 cases of CM. Preoperative routine pelvic anteroposterior film, entrance position, exit position and pelvic CT three-dimensional reconstruction were performed. Intraoperative O-arm navigation system three-dimensional reconstruction and triplane scanning imaging were used to evaluate the effect of intraoperative reduction. The anterior pelvic ring was fixed with internal external fixator, and the posterior ring was fixed with sacroiliac screw, plate screw or lumbar iliac screw. The operation time, intraoperative bleeding and nail placement were observed and recorded. The quality of fracture reduction was evaluated by Matta standard, and the postoperative function was evaluated by Majeed function score.@*RESULTS@#Wound healing was good in all patients without vascular, nerve and local irritation complications. All the 15 patients were followed up for 10 to 16 months. The fracture reduction was evaluated according to the Matta scoring standard, 9 cases were excellent results, 5 cases were good, and 1 case was medium. The Majeed functional score was 0 to 95 points.@*CONCLUSION@#The built-in external fixator assisted O-arm navigation imaging system in the treatment of unstable pelvic fractures. The reduction effect is evaluated in advance, the operation time is shortened, and the accuracy of internal fixation is improved. The operation is simple, safe and less bleeding. The operation is in line with the principles of minimally invasive medical treatment and precision medical treatment in orthopedics, which is conducive to the recovery of patients' postoperative function and rapid recovery.

Expression and refolding of OLA Ⅰ protein with peptides derived from sheeppox virus / 生物工程学报

The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.

Gene Mutation Types and Ethnic Distribution Characteristic of Thalassemia in Guiyang / 中国实验血液学杂志

OBJECTIVE@#To investigate the genetic mutation types and ethnic distribution characteristics of thalassemia in Guiyang.@*METHODS@#The population underwent physical examination, antenatal examination and pre-pregnancy examination in our hospital from January 2019 to November 2019 was selected, and the thalassemia gene was detected by RDB-PCR.@*RESULTS@#Among the 4 572 samples, 346 were positive , and the total carrying rate was 7.57%. The carrying rate of α-thalassaemia gene was 5.42% (248 cases), while β-thalassemia was 1.99% (91 cases), and α+β-compound thalassemia was 0.15% (7 cases). α-thalassaemia genotype with the lack of right side -α@*CONCLUSION@#The gene types of thalassemia in Guiyang are complex and varied. The -α

Relation of Blood Arsenic Concentration with Effect and Safety of Arsenic-Containing Qinghuang Powder () in Patients with Myelodysplastic Syndrome / 中国结合医学杂志

OBJECTIVE@#To investigate the relation of blood arsenic concentration (BAC) with clinical effect and safety of arsenic-containing Qinghuang Powder (, QHP) in patients with myelodysplastic syndrome (MDS).@*METHODS@#Totally 163 patients with MDS were orally treated with QHP for 2 courses of treatment, 3 months as 1 course. The BACs of patients were detected by atomic fluorescence spectrophotometry at 1, 3, and 6 months during the treatment, and the effective rate, hematological improvement and safety in patients after treatment with QHP were analyzed.@*RESULTS@#After 2 courses of treatment, the total effective rate was 89.6% (146/163), with 31.3% (51/163) of hematological improvement and 58.3% (95/163) of stable disease. The hemoglobin increased from 73.48 ± 19.30 g/L to 80.39 ± 26.56 g/L (P0.05). Among 46 patients previously depended on blood transfusion, 28.3% (13/46) completely got rid of blood transfusion and 21.7% (10/46) reduced the volume of blood transfusion by more than 50% after treatment. The BACs were significantly increased in patients treated for 1 month with 32.17 ± 18.04 μ g/L (P0.05). The adverse reactions of digestive tract during the treatment were mild abdominal pain and diarrhea in 14 cases (8.6%), and no patients discontinued the treatment. The BACs of patients with gastrointestinal adverse reactions were significantly lower than those without gastrointestinal adverse reactions (22.39 ± 10.38 vs. 37.89 ± 11.84, μ g/L, P<0.05). The BACs of patients with clinical effect were significantly higher than those failed to treatment (40.41 ± 11.69 vs. 23.84 ± 12.03, μ g/L, P<0.05).@*CONCLUSION@#QHP was effective and safe in the treatment of patients with MDS and the effect was associated with BACs of patients.

Relationship between TET2 Gene SNP rs3733609 C/T and JAK2V617F Allele Burden in Patients with Myeloproliferative Neoplasms / 中国实验血液学杂志

OBJECTIVE@#To investigate the relationship between the polymorphism of TET2 gene SNP rs3733609 and JAK2V617F allele burden in patients with myeloproliferative neoplasms (MPN).@*METHODS@#The exon 9 of TET2 gene was amplified by RT-PCR, and the nucleotide sequence of SNP rs3733609 site was analyzed by gene sequencing. The MGB Taqman probe PCR method was used to detect the JAK2V617F allele burden. The correlation of TET2 gene SNP rs3733609 C/T with the JAK2V617F allele burden and clinical parameters was analyzed.@*RESULTS@#TET2 gene rs3733609 C/T heterozygosity (normal T/T) could be detected in 19 cases of 85 cases of JAK2V617F positive MPN (22.4%) patients, while the TET2 gene rs3733609 C/T heterozygosity could be detected only in 9 of the 106 healthy volunteers, and the incidence was only 8.5% (9/106). Compared with the negative group (TET2 rs3733609 T/T), there was no significant difference in the median age, hemoglobin level and platelet count in the patients with TET2 gene SNP rs3733609 (CT/TC) positive, but the WBC count of peripheral blood and JAK2V617F allele burden significantly increased. In JAK2V617F high allele burden group, TET2 gene SNP rs3733609 was positive in 7 cases (36.8%, 7/19), the ratio was higher than that in the low allele burden group(18.2%, 12/66).@*CONCLUSION@#TET2 SNP rs3733609 C/T may be a new susceptible allelee, which affects the clinical characteristics and clonal evolution of MPN patients.

Association of Gene Mutations with Response to Arsenic-Containing Compound Qinghuang Powder () in Patients with Myelodysplastic Syndromes / 中国结合医学杂志

OBJECTIVES@#To investigate the relationship between gene mutations and response to Compound Qinghuang Powder (, CQHP) in patients with myelodysplastic syndrome (MDS).@*METHODS@#Forty-three MDS patients were genotyped by ultra-deep targeted sequencing and the clinical data of patients were collected and the relationship between them was analyzed.@*RESULTS@#Up to 41.86% of patients harbored genet mutations, in most cases with more than one mutation. The most common mutations were in SF3B1, U2AF1, ASXL1, and DNMT3A. After treatment with CQHP, about 88.00% of patients no longer required blood transfusion, or needed half of prior transfusions.@*CONCLUSIONS@#CQHP is an effective treatment for patients with MDS, especially those with gene mutations in SF3B1, DNMT3A, U2AF1, and/or ASXL1.

Secretory Imbalance between Pro-inflammatory and Anti-inflammatory Cytokines in the Patients with Immune Thrombocytopenia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the imbalance of pro-inflammatory and anti-inflammatory cytokines in the patients of immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Thirty-five patients with ITP were enrolled in ITP group, while 28 healthy persons were included in control group. The expressions of IL-8, IL-17A, IL-22, TNF-α, IFN-γ, IL-4, CD40, CD40L, TGF-β and IL-10 were detected by flow cytometry with aimPlex multiple immunoassay Flow.</p><p><b>RESULTS</b>The expressions of pro-inflammatory factors IL-8, IL-17A, IL-22, IFN-γ and TNF-α in ITP group all were significantly higher than that in the control group (P<0.05), however the expressions of anti-inflammatory factors IL-4, CD40L, TGF-β and IL-10 in ITP group all were significantly lower than that in the control group (P<0.05). The expression of CD40 was not significantly different between ITP group and control group (P>0.05). Expressions of TNF-α significantly related with platelet counts in both ITP group and control group (ITP group, r=0.64, P<0.05; control group, r=-0.41, P<0.05). However the expression of CD40, TGF-β, CD40L, IL-8, IL-17A, IL-22, IL-10, IL-1β, IFN-γ and IL-4 significantly did not relate with platelet counts in both ITP and control group.</p><p><b>CONCLUSIONS</b>The secretory imbalance between pro-inflammatory and anti-inflammatory cytokines exists in the patients of ITP. The decrease of Plt regulated may be regulated by the abnormal expression of TNF-α.</p>

Application of morcellator in 1.94 μm thulium laser enucleation of benign prostatic hyperplasia with volume >80 ml / 中国内镜杂志

Objective To investigate the effect of 1.94 μm thulium laser enucleation of benign prostatic hyperplasia (BPH) with volume >80 ml by morcellator. Methods From September 2014 to June 2016, there were 95 BPH patients with prostate volume over 80 ml treated by thulium laser were divided into two groups according to the surgical procedure: 45 cases in group A, prostate tissue were washed out of bladder after vapoenucleation by 1.94 μm thulium laser; 50 cases in group B, the enucleated prostate tissue were extracted by morcellator. The operation time, the decreasing level of hemoglobin on the first day after surgery, the hospitalization time, the gland tissue weight, catheterization duration, short-term incidence of complications, and the IPSS, PVP, Qmax, QOL in 3 months after surgeon of the two groups were observed and recorded. Results There was significant difference in operation time and gland tissue weight between the two groups. The group B have significantly short operation time compared with group A (P < 0.05), and obtained gland tissue remarkably exceed the group A (P < 0.05). No significant difference was found in hemoglobin level, hospitalization time, catheterization duration, and short-term complication between the two groups (P > 0.05). The IPSS, PVR, Qmax and QOL of 3 month, after operation were significantly improved but without any significant difference between the two groups (P < 0.05). Conclusion Vaporization cutting tissue or morcellating tissue after 1.94 μm thulium laser enucleation has high safety, good curative effect and low complication, while extraction prostate tissue by morcellator can shorten the operation time and get more tissues.

Prognostic Value of Multigene Methylation in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the methylation status of p15, DAPK, SOCS1 and FHIT genes in patients with myelodysplastic syndrome(MDS) and to explore the prognostic significance of gene methylation.</p><p><b>METHODS</b>Methylation-specific PCR (MSP) was used to detect the methylation of the above-mentioned 4 genes in 67 patients with MDS and 18 patients with iron-deficient anemia as controls. The gene methylation status of MDS patients and its effect on prognosis were analyzed.</p><p><b>RESULTS</b>The methylation rates of p15, DAPK, SOCS1 and FHIT in 67 MDS patients were 37.3%, 35.8%, 47.8% and 52.2%, respectively, which were significantly higher than those in the control group (P<0.05). The methylation status of p15, SOCS1 was increased along with the increase of International Prognostic Scoring System(IPSS) scores (P<0.05), and ≥2 genes was more frequent in relatively high risk groups (P<0.05). The median overall survival time of patients with and without methylation were 15 and 21 months, respectively (P<0.05). Patients showing methylation of SOCS1 had a significantly shorter survival time in relatively low risk groups(P<0.05), meanwhile SOCS1, p15 and methylations of ≥2 genes had significantly shorter survival time in relatively high risk groups(P<0.05). In multivariate analysis, SOCS1 and p15 were negative prognostic factors.</p><p><b>CONCLUSION</b>p15, DAPK, SOCS1 and FHIT are higher hypermethylated genes in MDS. The methylations of SOCS1 and p15 are independent prognostic factor for overall survival in MDS.</p>

Clinical Value of p15, DAPK, SOCS1 and FHIT Genes Combined Detection in the Early Diagnosis and Prognosis Evaluation of Myelodysplastic Syndrome / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the value of p15, DAPK, SOCS1 and FHIT genes combined detection in the early diagnosis and prognosis evaluation of patients with myelodysplastic syndrome(MDS).</p><p><b>METHODS</b>The methylation-specific PCR (MSP) was used to detect the methylation of the above-mentioned 4 genes in 67 patients with MDS. The value of 4 gene combined detection in the early diagnosis and prognosis evaluation of patients with MDS was compared and anazlyzed.</p><p><b>RESULTS</b>The methylation rates of p15, DAPK, SOCS1 and FHIT genes in 67 patients with MDS were 37.3%, 35.8%, 47.8% and 52.2%, respectively, which were significantly higher than those in control group (P<0.05). The accordance rates of p15, DAPK, SOCS1 and FHIT single detection for diagnosis of MDS were 37.3%,35.8%,47.8% and 52.2%, respectively, meanswhile the accordance rate of above-mentioned 4 gene combined detection for diagnosis of MDS was 82.1%, which was significantly higher than that of single gene detection(P<0.001). The methylation of ≥2 genes in relatively high risk group was significantly higher than that in relatively low risk group (P<0.05). The median survival time of MDS patients was 18(13.3, 22.7) months; the median survival time in relatively low risk group was significantly longer than that in relatively high risk group [27(20.3,33.7) months vs 9(5.9,12.1) months] (P<0.05). The survival time of MDS patients with different risks displayed the trend of shorting feature along with increasing of methylated genes (P<0.05).</p><p><b>CONCLUSION</b>The combined detection of above menthioned 4 genes can improve the accuracy of early diagnosis and prognosis evaluation for MDS patients.</p>

Regulative effect of Xuanbai Chengqi Decoction on TNF-α and IL-1β in patients with acute respiratory distress syndrome and its mechanical indexes / 实用医学杂志

Objective To study the effects of Xuanbaichengqi decoction on the regulation of inflammatory factors TNF-α and IL-1 β in patients with acute respiratory distress syndrome (ARDS) and its mechanism.Methods 60 patients with ARDS who had been hospitalized from January 2016 to August 2016 were randonly selected and then assigned to a control group (30 patients) and a study group (30 patients).Both groups received routine therapies and mechanical ventilation,and the study group received additional Xuanbaichengqi decoction.Levels of TNF-o,IL-1β and IL-10,and peak inspiratory pressure (PIP),plateau airway pressure (Pplat),PVC,forced vital capacity (FVC),forced expiratory volume in one second (FEV 1),and oxygenation index (PaO2/FiO2) were detected before and after treatment.Results Levels of TNF-α and IL-1βwere significantly lower in the study group than in the control group (P ＜ 0.05).IL-10 level was significantly higher in the study group than in the control group (P ＜ 0.05).And the improvement in PIP and Pplat was also significant in the study group (P ＜ 0.05).Less adverse reactions occurred in the study group.Conclusions Xuanbaichengqi decoction can improve the indexes for respiratory mechanics and pulmonary function in patients on mechanical ventilation and reduce the inflammatory response,having a significant effect.

Study of polymorphisms of genes related to folic acid metabolism among women of child-bearing age from Shanxi / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze polymorphisms of genes related to folic acid metabolism among women of child-bearing age from Shanxi.</p><p><b>METHODS</b>Buccal smears were collected from 1070 women of child bearing age with cotton swabs. Sequences of MTHFR C667T and A1298C, MTRR A66G, and SLC19A1 A80G were determined by DNA sequencing. The results were compared with data from other regions of China.</p><p><b>RESULTS</b>For MTHFR C667T, the wild type homozygote, heterozygous mutants, and homozygous mutants have respectively accounted for 20.5%, 50.3%, and 29.2% of the study group, with the frequency of the mutant T allele being 54.4%. For MTHFR A1298C, these were 68.7%, 29.3%, and 2.0%, with the frequency of mutant C allele being 16.6%. For MTRR A66G, the above frequencies were 51.5%, 41.8%, and 6.7%, with the frequency of the mutant G allele being 27.6%. For SLC19A1 A80G, these were 29.2%, 48.0%, 22.8%, with the frequency of mutation G allele being 46.8%. Compared with other regions of China, women of child-bearing age from Shanxi has shown a significant difference in allelic distribution of MTRR A66G and SLC19A1 A80G (P<0.05).</p><p><b>CONCLUSION</b>The polymorphisms of genes related to folic acid metabolism showed significant regional difference. Over half of women from Shanxi have carried high-risk alleles for folic acid insufficiency and should have individualized folic acid supplement.</p>

Mechanisms of total flavones of Epimedium on oxidative stress induced by myocardial ischemia/reperfusion injury in rats / 中国中药杂志

To investigate the effects and mechanisms of total flavones of Epimedium (TFE) on oxidative stress induced by myocardial ischemia/reperfusion injury in rats, forty male SD rats were randomly divided into sham operated group, model group, diltiazem group and flavonoids of Epimedium low and high doses groups with 8 rats in each. Myocardial ischemia/reperfusion injury model was induced by ligaturing the left anterior descending artery for 30 min followed reperfusion for 4 h after TFE was taken by intragastric administration for 4 days. The degree of myocardial infarct was observed by N-BT staining. The concentrations of MDA and activities of SOD and T-AOC in cardiac tissue were measured by colorimetry. Serum TnI concentrations were checked by ELISA. HE stain was used to observe myocardium structure under light microscope. Expressions of SIRT1 and Nrf2 in cardiac tissue were evaluated by immunohistochemistry method and Western blot, respectively. Compared with the model group, the degree of myocardial infarct, MDA concentration in cardiac tissue and the levels of TnI in serum significantly decreased in the diltiazem group and flavonoids of Epimedium low and high doses groups (P<0.05 or P<0.01); flavonoids of Epimedium low and high doses groups and the diltiazem group also showed improvements in myocardium structure under ischemia/reperfusion injury. TFE significantly increased the activity of SOD and T-AOC and the expression of SIRT1 and Nrf2 in cardiac tissue when compared with the model group (P<0.05 or P<0.01). Therefore, TFE can increase anti-peroxidant capacity of myocardium tissue by using intrinsically anti-oxidant signaling pathway of SIRT1 and Nrf2, which can inhibit irreversible damage of cardiomyocytes in myocardial ischemia/reperfusion injury and protect normal function of cardiac tissue.

LC-MS/MS method for the simultaneous quantitative analysis of dihydromyricetin and myricetin in mice plasma and its application to a tissue distribution study / 中国药学杂志

OBJECTIVE: To establish an LC-MS/MS method for the simultaneous quantitative analysis of dihydromyricetin and myricetin in mice plasma, and for application in tissue distribution study after oral administration dihydromyricetin(DMY) and myricetin(MY). METHODS: All groups of mice were sacrificed and the heart, liver, spleen, lung, kidney, brain and plasma were collected after oral administration of DMY and MY at the dose of 144.4 mg·kg-1, respectively. The protein precipitation approach with acetonitrile containing 0.1% formic acid was adopted for the sample preparation. The liquid chromatographic separation process was performed with an Ultimate® AQ-C18(3.0 mm×100 mm, 3.0 μm) to determine the concentrations of DMY and MY in all tissue, and then all data were analysed. RESULTS: It was proved that the accuracy, precision, matrix effect and extract recovery of this method can meet the requirements of biological sample analysis. The RESULTS indicated that DMY and MY were quickly absorbed in the body. The maximum concentrations of DMY and MY in the liver were higher than those in the other tissues. Moreover, the levels of DMY in plasma and other tissues were higher than MY. CONCLUSION: A rapid, reliable, sensitive LC-MS/MS method for the simultaneous quantification of DMY and MY is established in the present research. Meanwhile, the result of the tissue distribution study will provide a reference for the reasonable development and the future clinical applications of DMY and MY.

Diagnostic value of high-risk human papillomavirus, p16 and Ki-67 detection in the cervical intraepithelial neoplasia / 肿瘤研究与临床

Objective To investigate the expression of HPV-DNA and p16,Ki-67 proteins in intraepithelial lesion of cervix (CIN) so as to provide the aluable evidence for the early clinical diagnosis and genetic therapy of the CIN.Methods Hybrid capture2 was used to detect HPV DNA in 35 cases of CIN Ⅰ,44 cases of CIN Ⅱ,and 20 cases of CIN Ⅲ.Immunohistochemical MaxVision was used to research the expression of p16 and Ki-67 protein in CIN Ⅰ,CIN Ⅱ and CIN Ⅲ.The results were analyzed with clinicopathological factors.Results Positive expression rates of HPV-DNA were 31.4 ％ (11/35),63.6 ％ (28/44),85.0 ％ (17/20) in CIN Ⅰ,CIN Ⅱ and CIN Ⅲ,respectively.The expression rates of p16 in the three groups were 65.7 ％ (23/35),97.7 ％ (43/44),100.0 ％ (20/20),there were statistically significant difference (x2 =21.30,P =0.00).The expression rates of Ki-67 in the three groups were 77.1％ (27/35),79.5 ％ (35/44),95.0 ％ (19/20),there were no difference (x2 =3.00,P =0.22).Conclusions HPV infection is closely related to the occurrence and development of CIN.p16,Ki-67 can be used as objective marks in diagnosis of CIN.

LC-MS/MS method for determination of sixteen bile acids in rat plasma and its application in liquorice detoxification research / 中国药学杂志

OBJECTIVE: To establish an LC-MS/MS method for determination of the concentrations of 16 bile acids in rat plasma. METHODS: The plasma protein was precipitated with methanol and then the supernatant was dried. Liquid chromatographic separation was performed on a Xtimate™ C18 column with gradient elution of mobile phase consisting of methanol-ammonium acetate and formic acid buffer, the flow rate was 0.25 mL · min-1, and the injection volume was 5 μL. RESULTS: The calibration curves were linear in the concentration range of 0.0056-3.8000 μmol · L-1. The lower limit of quantification was 0.005 6μmol · L-1. The recoveries of the samples at low, middle and high concentrations were 70.3%-107.2%. The intra-day and inter-day RSDs were less than 12.9%. CONCLUSION: This method is accurate and sensitive for determination of bile acids in rat plasma, which provides reference for the determination of bile acids in clinical samples.

Evaluation on hepatoprotection in Nrf2/ARE oxidation/chemical stress defense pathway caused by isoliquiritigenin based on analysis of bile acids / 中国药学杂志

OBJECTIVE: To evaluate hepatoprotection in Nrf2/ARE oxidation/chemical stress defense pathwaycaused by isoliquiritigenin (Iso) based on analysis of bile acids using profile of bile acids. METHODS: High performance liquid chromatography coupled with quadrupole mass spectrometry (HPLC-MS/MS) was applied to determine the contents of all kinds of endogenous bile acids including free bile acids, taurine conjugates and glycine conjugates. By the luciferase reporter gene assay for screening Nrf2-ARE signal targeting molecular experiments in liquorice extract, isoliquiritigenin was capable of markedly activating Nrf2-driven gene expression, therefore it was used to evaluate hepatoprotection in Nrf2/ARE oxidation/chemical stress defense pathway. RESULTS: Based on the analysis using principle components analysis (PCA), partial least square-discriminant analysis (PLS-DA), Nrf2+/+, Nrf2-/- and Iso Nrf2+/+ groups could be distinguished from their Iso Nrf2-/- group, which suggested that the variance of the contents of bile acids could evaluate hepatoprotection caused by Iso. Bile acids of ursodeoxycholic acid (UDCA), chenodeoxycholic acid (CDCA), cholic acid(CA), sodium taurodeoxycholate hydrate (TDCA), deoxycholic acid (DCA), taurocholic acid(TCA) proved to be important corresponds to Iso induced liver protection according to analysis of partial least square-discriminant analysis (PLS-DA) and the statistical analysis showed that there were significant differences among the four groups. It indicated that UDCA, CDCA, CA, TDCA, DCA and TCA could be considered as sensitive biomarkers of Iso induced liver protection. CONCLUSION: This work can provide the base for the further research on the evaluation and mechanism of hepatoprotection caused by liquorice.

Determination of diclofenac sodium in human plasma based on 2D-HPLC coupled with trapping column and its application / 中国药学杂志

OBJECTIVE: To establish a method for determination of diclofenac sodium in human plasma under high-fat meal condition using two-dimensional liquid chromatography (2D-HPLC) coupled with trapping column, and to evaluate the bioequi valence and the bioavailability of diclofenac sodium sustain-released tablets in healthy volunteers. METHODS: Under fed state, eighteen healthy male volunteers were divided into two groups by an open, randomized two period crossover design with a single dose of diclofenac sodium sustain-released tablets. The plasma concentrations were determined by 2D-HPLC method, and calculated pharmacokinetic parameters and bioavailability. RESULTS: The main pharmacokinetic parameters of test and reference preparations after a single dose were: AUC0-16 was (2 591.6 ± 705.8) and (2 588.8 ± 772.0) ng · h · mL-1; AUC1→∞ was (2 896.4 ±839.7) and (2 700.3 ± 806.1) ng · h · mL-1; tmax was (4.6 ± 0.7) and (4.4 ± 0.9)h; ρmax was(1 332.8 ± 912.5) and (1 325.7 ± 706.3) ng · mL-1, respectively. The relative bioavailability of test preparation in single dose was (102.4 ± 15.1)%. CONCLUSION: The 2D-HPLC method coupled with trapping column is a simple, rapid and specific for determination of diclofenac sodium in human plasma. The results of statistical analysis indicate that the two preparations are bioequivalent in healthy volunteers with a single dose under high-fat fed condition.