Study on quality control method of the roots and rhizoma of Toricellia angulata / 中国药房

OBJECTIVE To establish the quality control method for the roots and rhizoma of Toricellia angulata. METHODS The properties of the roots and rhizoma of T. angulata were observed and microscopic identification was conducted. The moisture， total ash， acid-insoluble ash and ethanol-soluble extract were examined according to the method stated in the 2020 edition of Chinese Pharmacopoeia （part Ⅳ）. HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata were established， common peaks were identified and the similarity was evaluated by using the Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）. The contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol were determined by HPLC. RESULTS The properties and microscopic identification of the roots and rhizoma of T. angulata were obvious. The average contents of moisture， total ash， acid-insoluble ash and ethanol-soluble extract were 7.54%， 2.18%， 0.15% and 7.81%， respectively. There were 16 common peaks marked in the HPLC fingerprints of 11 batches of the roots and rhizoma of T. angulata， with similarities of 0.856-0.960； five of them were identified， such as coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D-glucopyranoside and syringaresinol. The contents of the above five components were 0.047 2-0.401 6， 0.836 8-8.697 9， 1.245 3-10.950 0， 0.139 0-0.437 8 and 0.016 4-0.635 3 mg/g， respectively. CONCLUSIONS The established method is stable and accurate， which can be used for the quality control of the roots and rhizoma of T. angulata. It is preliminarily proposed that the moisture in the roots and rhizoma of T. angulata is not more than 11.0%， the total ash is not more than 4.0%， the ethanol-soluble extract is not less than 5.0%， the contents of coniferin， syringin， chlorogenic acid， （+）-syringaresinol-O-β-D- glucopyranoside and syringaresinol are not less than 0.04，0.83， 1.24， 0.13， 0.01 mg/g， respectively.

Comparative study on in vivo and in vitro permeability of Huoxue Zhitong gel patch and microemulsion gel / 中国中药杂志

This study aims to establish a method for determination of paeonol(Pae), eugenol(Eug), and piperine(Pip) content in receptor liquid and research on the permeability and pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The Franz diffusion experiment was conducted to assess the percutaneous permeability, and the microdialysis method was employed to assess pharmacokinetics of Huoxue Zhitong gel patch and microemulsion gel. The content of Pae, Eug, and Pip in receptor liquid in vitro and in vivo was determined by HPLC and UPLC-MS. The Q_n and J_(ss) of Pae, Eug, and Pip in the gel patch were significantly higher than those in the microemulsion gel, indicating that the drug release was faster in the gel patch. The C_(max), AUC_(0-760), and MRT of Pae, Eug, and Pip in the gel patch were higher than those in the microemulsion gel, indicating that the gel patch can promote the penetration and prolong the skin residence of the drug. The results of this study provide reference for improving the dosage form of Huoxue Zhitong patch.

Effective Components of Periploca forrestii Against Rheumatoid Arthritis by Targeting TNF-α / 中国实验方剂学杂志

ObjectiveTo analyze the effective components of Periploca forrestii against rheumatoid arthritis（RA）by targeting tumor necrosis factor （TNF）-α based on network pharmacology and experimental verification. MethodThe preliminary research of the research group found that the alcohol extracts of P. forrestii （CDLF and CQAF） had significant anti-RA activities，and 10 monomers with such activities were identified. The anti-RA activities of active monomers，CDLF， and CQAF were compared by the enzyme-linked immunosorbent assay （ELISA）with interleukin（IL）-6，nitric oxide （NO），IL-1β， and prostaglandin E2（PGE2）as indicators. Network pharmacology was employed to analyze the possible molecular mechanism of P. forrestii against RA. The targeting ability of P. forrestii chemical monomers to TNF-α was verified by TNF-α molecular docking，surface plasmon resonance （SPR）， and TNF-α-induced L929 injury model. ResultELISA showed that the anti-RA activities of CDLF and CQAF were significantly stronger than those of identified 10 active monomers. Network pharmacology analysis showed that the core targets of P. forrestii against RA were signal transducer and activator of transcription protein 3 （STAT3），TNF， and IL-6. Gene Ontology（GO） analysis revealed collagen catabolism，inflammatory response，positive regulation of nuclear factor kappa-B（NF-κB） transcription factor activity，and positive regulation of B cell proliferation. Kyoto Encyclopedia of Genes and Genomes （EKGG） pathway enrichment analysis demonstrated TNF signaling pathway，phosphoinositide 3-kinase（PI3K）/protein kinase B（Akt） signaling pathway，NF-κB signaling pathway，Toll-like receptor signaling pathway，mitogen-activated protein kinase（MAPK） signaling pathway， etc. Verification experiments by TNF-α molecular docking，SPR， and TNF-α-induced L929 injury model found that CDLF and CQAF had good binding activities and could manifestly antagonize TNF-α. However， the active components separated and identified from CDLF and CQAF did not show the same anti-TNF-α activity. ConclusionThe CDLF and CQAF of P. forrestii may treat RA by targeting TNF-α. The experiments found that the isolated chemical components had weaker binding activity to TNF-α than CDLF and CQAF. Meanwhile，the research group isolated chemical components with a minimum mass fraction of 0.25 ng·g-1 from P. forrestii， which suggested that the active components generated by binding to TNF-α with anti-RA activities were presumedly trace components .

Establishment of Flp-In CHO recombinant cell lines stably expressing CYP2A13 and CYP2A13/POR / 中国药理学通报

Aim To construct Flp-In CHO cell line(CYP2A13-CHO)stably expressing cytochrome P450 family 2 subfamily A member 13(CYP2A13)and Flp-In CHO cell line(CYP2A13-POR-CHO)stably co-expressing CYP2A13 and cytochrome P450 oxidoreductase(POR), from which a cell line with better metabolic activity is selected.Method In our previous study, we had constructed a Flp-In CHO cell line(POR-Flp-In CHO)stably expressing POR using lentiviral vector.The recombinant plasmids of pcDNA5/FRT-CYP2A13 were constructed and transfected into Flp-In CHO cells and POR-Flp-In CHO cells through LipofectamineTM 2000.The expression and activity of CYP2A13 were detected by real-time quantitative PCR(qRT-PCR), Western blot and Aflatoxin B1(AFB1)/4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)cytotoxicity assay and the metabolic activity was compared between CYP2A13-CHO and CYP2A13-POR-CHO.Results Compared with non-transfected cells, the mRNA and protein expression of CYP2A13 in CYP2A13-CHO and CYP2A13-POR-CHO cells both increased significantly.Besides, compared with CYP2A13-POR-CHO, CYP2A13-CHO cells were more sensitive to AFB1 and NNK.Conclusions The Flp-In CHO cell line stably expressing CYP2A13 and with better metabolic activity has been established successfully, which provides a tool for screening of pre-carcinogens that can be metabolically activated by CYP2A13.

Clinicopathological characteristics and prognosis of patients with sporadic multiple primary gastrointestinal stromal tumors / 中华胃肠外科杂志

Objective: To investigate the clinicopathological characteristics and prognosis of sporadic multiple primary gastrointestinal stromal tumor (GIST). Methods: A retrospective cohort study was conducted. Case inclusion criteria: (1) postoperative pathological diagnosis of GIST; (2) primary GIST with single lesion or sporadic multiple primary GIST (sporadic GIST was defined as primary GIST other than familial and syndrome-related GIST, and multiple primary GIST was defined as the number of primary GISTs in the same patient ≥ 2); (3) patients with complete clinicopathological data. Those with tumor recurrence or distant metastasis, and with other malignancies were excluded. Medical records of patients with primary GIST who underwent surgical resection in the Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 2010 to December 2020 were collected. Patients were divided into sporadic multiple primary GIST group and single primary GIST group according to the number of primary GIST lesions. The clinicopathological data and prognosis of the two groups were observed and compared. Results: A total of 1200 patients with primary GIST were enrolled in this study, including 628 males (52.3%) and 572 females (47.7%), with a median onset age of 58 (19-93) years. Among them, 1165 cases (97.1%) were sporadic primary GIST with single lesion; 35 cases (2.9%) were sporadic multiple primary GIST. Among 35 cases of sporadic multiple primary GIST, 3 cases (8.6%) had acid reflux as the first symptom, which was higher than the single primary GIST group (22/1165, 1.9%) (χ(2)=7.437, P=0.006). There were no significant differences in other clinical characteristics between the two groups (all P>0.05). Patients in the sporadic multiple primary GIST group contained a total of 80 primary tumors. Compared with the single primary GIST group, the sporadic multiple primary GIST group had a higher proportion of tumors originating in the stomach [87.5% (70/80) vs. 59.1% (689/1165)], lower proportion of spindle cell in histology [85.0% (68/80) vs. 93.7% (1092/1165)], higher proportion of positive CD34 [97.5% (78/80) vs. 87.6% (1021/1165)], smaller maximum diameter [maximum diameter ≤2.0 cm: 61.2% (49/80) vs. 28.8% (335/1165)], lower mitotic rate [≤5/50 high-power fields (HPF): 93.8% (75/80) vs. 74.5% (868/1165)], lower risk of recurrence [60.0% (48/80) vs. 23.3% (271/1165)], and the differences were all statistically significant (all P<0.05). The 3-year recurrence-free survival rate in the sporadic multiple primary group and the single primary GIST group was 96.6% and 89.3% respectively (P=0.160), and the 3-year overall survival rate was 100.0% and 92.8%, respectively (P=0.088). Conclusions: The most common type of sporadic multiple primary GIST is multiple tumors originating in the stomach at the same time. Compared with primary GIST with single lesion, sporadic multiple primary GIST presents smaller maximum diameter and lower mitotic rate. The prognosis of patients between two groups is not significantly different.

Adherence to adjuvant with therapy imatinib in patients with gastrointestinal stromal tumor: a national multi-center cross-sectional study / 中华胃肠外科杂志

Objective: To analyze the current adherence to imatinib in patients with gastrointestinal stromal tumors (GIST) in China and its influencing factors. Methods: A cross-sectional survey was conducted. Study period: from October 1, 2020 to November 31, 2020. Study subjects: GIST patients taking imatinib who were diagnosed and treated in public tertiary level A general hospitals or oncology hospitals; those who had not been pathologically diagnosed, those who never received imatinib, or those who had taken imatinib in the past but stopped afterwards were excluded. The Questionnaire Star online surgery platform was used to design a questionnaire about the adherence to adjuvant imatinib therapy of Chinese GIST patients. The link of questionnaire was sent through WeChat. The questionnaire contained basic information of patients, medication status and Morisky Medication Adherence Scale. Results: A total of 2162 questionnaires from 31 provinces, autonomous regions, and municipalities were collected, of which 2005 were valid questionnaires, with an effective rate of 92.7%. The survey subjects included 1104 males and 901 females, with a median age of 56 (22-91) years old. Working status: 609 cases (30.4%) in the work unit, 729 cases (36.4%) of retirement, 667 cases of flexible employment or unemployment (33.3%). Education level: 477 cases (23.8%) with bachelor degree or above, 658 cases (32.8%) of high school, 782 cases (39.0%) of elementary or junior high school, 88 cases (4.4%) without education. Marital status: 1789 cases (89.2%) were married, 179 cases (8.9%) divorced or widowed, 37 cases (1.8%) unmarried. Two hundred and ninety-four patients (14.7%) had metastasis when they were first diagnosed, including 203 liver metastases, 52 peritoneal metastases, and 39 other metastases. One thousand eight hundred and sixty-nine patients underwent surgical treatment, of whom 1642 (81.9%) achieved complete resection. The median time of taking imatinib was 25 (1-200) months. Common adverse reactions of imatinib included 1701 cases (84.8%) of periorbital edema, 1031 cases (51.4%) of leukopenia, 948 cases (47.3%) of fatigue, 781 cases (39.0%) of nausea and vomiting, 709 cases (35.4%) of rash, and 670 cases (33.4%) of lower extremity edema. The score of the Morisky Medication Adherence Scale showed that 392 cases (19.6%) had poor adherence, 1023 cases (51.0%) had moderate adherence, and 590 cases (29.4%) had good adherence. Univariate analysis showed that gender, age, work status, economic income, residence, education level, marriage, the duration of taking medication and adverse reactions were associated with adherence to adjuvant imatinib therapy (all P<0.05). Multivariate analysis showed that female (OR=1.264, P=0.009), non-retirement (OR=1.454, P=0.001), monthly income ≤4000 yuan (OR=1.280, P=0.036), township residents (OR=1.332, P=0.005), unmarried or divorced or widowed (OR=1.362, P=0.026), the duration of imatinib medication >36 months (OR=1.478, P<0.001) and adverse reactions (OR=1.719, P=0.048) were independent risk factors for poor adherence to adjuvant imatinib. Among patients undergoing complete resection, 324 (19.7%) had poor adherence, 836 (50.9%) had moderate adherence, and 482 (29.4%) had good adherence. Meanwhile, 55 patients with good adherence (11.4%) developed recurrence after surgery, 121 patients with moderate adherence (14.5%) developed recurrence, 61 patients with poor adherence (18.8%) developed recurrence, and the difference was statistically significant (P=0.017). Conclusions: The adherence to adjuvant therapy with imatinib in Chinese GIST patients is relatively poor. Females, non-retirement, monthly income ≤4000 yuan, township residents, unmarried or divorced or widowed, the duration of imatinib medication >36 months, and adverse reactions are independently associated with poor adherence of GIST patients. Those with poor adherence have a higher risk of recurrence after surgery. Positive interventions based on the above risk factors are advocated to improve the prognosis of patients with GIST.

Effects of intermittent hypoxia stimulation with different frequencies on HT22 cell viability and expression of Hif-1α and p-NF-κB / 生理学报

Intermittent hypoxia (IH) could induce cognitive impairment through oxidative stress and inflammation. However, the degree of cell damage is closely related to the IH stimulus frequency. IH stimulation with different frequencies also induces opposite results on neuronal cell lines. Therefore, this study was aimed to compare the effects of IH stimulation with three different frequencies on murine hippocampal neuronal HT22 cell activity, and to explore the molecular mechanism of the IH stimulus frequency-related neuron injury. HT22 cells were cultured and divided into control group and three IH stimulation groups with different frequencies. Oxygen concentration in the chamber was circulated between 21% and 1% (IH1 group, 6 cycles/h; IH2 group, 2 cycles/h; IH3 group, 0.6 cycle/h). Cell morphology was observed at 6, 12, 24 and 48 h of IH treatment. Cell viability was determined by the CCK-8 kit, lactate dehydrogenase (LDH) content in cell supernatant was determined by LDH kit, oxidative stress level was detected by the reactive oxygen species (ROS) probe, and protein expression levels of hypoxia inducible factor-1α (Hif-1α) and phosphorylated nuclear factor κB (p-NF-κB) were detected by Western blot. The results showed that, compared with control group, cell number and activity in the three IH groups were decreased, LDH content and ROS levels were increased with the prolongation of IH stimulation time, and the changes were most obvious in the IH1 group among those of the three IH groups. Hif-1α expression and the p-NF-κB/NF-κB ratio were also up-regulated with the prolongation of IH stimulation time, and the changes of IH1 group were the most significant. These results suggest that IH stimulation induces oxidative stress injury in HT22 cells, which is related to increased Hif-1α expression and NF-κB phosphorylation. Moreover, the higher frequency of IH stimulation induces more serious cell injury.

Research progress on chemical composition and pharmacological effects of Periplocae Cortex and predictive analysis on Q-marker / 中国中药杂志

Periplocae Cortex is a traditional Chinese medicine in China, which is mainly produced in northeast China, north China, northwest China, southwest China. In recent years, the increasing in-depth research resulted in the discovery of anti-tumor and cardiac pharmacological activities of Periplocae Cortex, which has broad application prospects. On the basis of summarizing chemical components and pharmacological effects, combined with the theoretical system of Q-marker, the quality control components of Periplocae Cortex were predicted from the aspects of the correlation between chemical composition and traditional medicinal properties, traditional efficacy, and new clinical use, plasma composition, measurable composition, storage time by analyzing literature. Among the components, periplocoside, periplocin, periplogenin, 4-methoxy salicylaldehyde showed significant activity, which provides a scientific basis for quality evaluation of Periplocae Cortex.

Analysis of the correlation between the extreme temperature of incision surface and non-union of incision in internal fixation of calcaneal fracture / 中国基层医药

To explore the correlation between the extreme temperature of incision surface and non-union of incision in internal fixation of calcaneal fracture.Methods From January 2016 to December 2017, 142 patients with closed unilateral calcaneal fracture treated with open reduction and internal fixation in the Integrated Chinese and Western Medicine Hospital of Wenzhou were enrolled.The incision temperature was collected by infrared radiation thermometers intermittently for 4 -5 times, and the extreme values were used for further analysis.The primary endpoint of the study was the incision healing 14 days after surgeries.The patients′age,gender,inflammatory indicators and body mass index (BMI) were evaluated,as well as related factors such as diabetes and arteriosclerosis. Results All patients completed the follow -up observation 14 days after operation,and finally achieved the wound healing in 105 cases,and the wound non -healing in 37 cases.The average extreme value of low temperature was (34.91 ±0.47)℃in the union group and (34.13 ±0.60)℃in the disunion group,and the average extreme value of high temperature was (36.61 ±0.55)℃in the union group and (36.90 ±0.52)℃in the disunion group,there were statistically significant differences between the two groups ( t ＝7.152,-2.762, P ＝0.000, 0.007 ).Logistic regression analysis results showed that age [β(regression coefficient )＝-0.111],extreme high( β＝2.728) or very low (β＝-3.854 ), arteriosclerosis ( β＝4.597 ), history of diabetes ( β＝2.234 ) were risk factors for incision nonunion(P＝0.046,0.003,0.001,0.002,0.024).Conclusion Extreme incision temperature is observed to be associated with an increased risk of operative incision disunion after open reduction and internal fixation for calcaneal fractures.

Electroacupuncture ameliorated neuropathic pain induced by chronic constriction injury via inactivation of PI3K/AKT pathway

@#Objective: To explore the analgesic effect of electroacupuncture on neuropathic pain and the role of PI3K/AKT pathway in microglial activation in spinal cord. Methods: Chronic constriction injury model was established using Sprague-Dawley rats to mimic neuropathic pain. The pain intensity was then detected by paw withdrawal threshold and thermal withdrawal latency. Real-time quantitative PCR (qRT-PCR), Western blot were used to evaluate the activation of PI3K/AKT pathway as well as microglia in L4-5 of all rats. Proinflammatory cytokines TNF-α and IL-6 were analyzed by Enzyme-linked immunosorbent assay (ELISA). Results: It was demonstrated that chronic constriction injury-induced hyperalgesia was significantly improved by LY294002 intrathecal administration and electroacupuncture stimulation at “Zusanli” 足三里 (ST36) and “Yanglingquan” 阳陵泉 (GB34). Also, treatment with electroacupuncture significantly reduced the activation of microglia and downregulated the levels of TNF-α and IL-6, which was similar to the outcomes of LY294002 intrathecal administration. Furthermore, the expression and phosphorylation of PI3K/AKT signaling was markedly suppressed by electroacupuncture treatment. Conclusions: These findings indicated that electroacupuncture exhibited the analgesic effect on CCI rats by inhibiting the activation of microglia and production of proinflammatory cytokines in spinal cord through blocking the microglial PI3K/AKT signaling activation

Study on the mechanism of intense pulsed light therapy for evaporative dry eye disease / 国际眼科杂志(Guoji Yanke Zazhi)

@#Dry eye disease(DED)is closely related to inflammatory diseases of the skin near the eyelids, such as ocular rosacea, <i>etc</i>. At present, intense pulsed light(IPL)is one of the main treatments for rosacea. Recent studies have found that patients with meibomian gland dysfunction(MGD)have alleviated the symptoms and signs of dry eyes during IPL treatment, but its specific mechanism is not clear. The purpose of this paper is to review the possible mechanisms of IPL treatment of dry eye and to discuss its rationality.

Preparation and Quality Evaluation of Pogostone Transfersomes / 中国药房

OBJECTIVE： To prepare pogostone transfersomes， and to evaluate its quality. METHODS： Film dispersion method was used to prepare pogostone transfersomes. Using the accumulative penetration volume （Qn） and accumulative penetration ratio （PR） of pogostone as evaluation indexes， the types of surfactant， formulation were screened in respects of the dosage of surfactant and the dosage of pogostone. The pogostone transfersomes were prepared with optimal formulation； the morphology， particle size distribution and Zeta potential were observed and the entrapment efficiency was measured. RESULTS： The optimal formulation was as follows as the sodium cholate was selected as surfactant； the dosage of sodium cholate was 0.25 g； the dosage of pogostone was 15 mg. The optimal pogostone transfersomes were ivory-white suspension； average particle size was （115.6±3.65） nm （RSD＝3.20％，n＝3）； PDI was 0.185±0.008 （RSD＝4.30％， n＝3）； Zeta potential was （－13.76±0.225） mV （RSD＝1.70％，n＝3）； entrapment efficiency of pogostone was （46.01±0.40）％ （RSD＝0.87％，n＝3）； Qn was （378.76±0.61）   μg/cm2 （RSD＝0.20％，n＝3）； PR was （89.02±0.96）％ （RSD＝1.10％，n＝3）. CONCLUSIONS： Prepared pogostone transfersomes are in line with quality requirements， which can provide reference for the further study of new dosage form of pogostone.

Site-targeted imaging enhancement of viable myocardium after ischemia-reperfusion by a novel nano-scale ultrasound contrast agent: a vivo study / 中华超声影像学杂志

Objective@#To prepare a kind of lipid nanoparticle ultrasound contrast agents with the ability to target to viable myocardium for diagnosis.@*Methods@#The agent was a biotinylated, fluorescent-labelled, lipid-coated, liquid perfluorocarbon emulsion. Physico-chemical properties of the agent were measured, including size distribution, Zeta Potential, concentration and so on. Ischemia-reperfusion models were created in rats, and then exposed to biotinylated anti-MCP-1 monoclonal antibody, rhodamine avidin and biotinylated, FITC-labelled nanoparticles, respectively. Echocardiography was taken before and after injection. Frozen sections of their hearts were observed under fluorescence microscope.@*Results@#The particle diameter, zeta potential and concentration of lipid nanoparticles were (172.30±52.06)nm, (-33.10±6.50)mV and (2.28±0.46)×1011/ml, respectively. From the short-axis view, the myocardium under endocardium of anterior wall was enhanced obviously. While myocardium of other walls were still. The lipid nanoparticles located in the myocardium of anterior wall and gave out bright green and red fluorescence under fluorescence microscope, while neither lipid nanoparticles nor fluorescence were found in other sites of ventricular myocardium.@*Conclusions@#The viable myocardium can be targeted and acoustically enhanced by the self-made nano-scale ultrasound contrast agent. This new agent has potential to improve sensitivity and specificity for noninvasive identifying viable myocardium.

Site‐targeted imaging enhancement of viable myocardium after ischemia‐reperfusion by a novel nano‐scale ultrasound contrast agent :a vivo study / 中华超声影像学杂志

Objective To prepare a kind of lipid nanoparticle ultrasound contrast agents with the ability to target to viable myocardium for diagnosis . Methods T he agent was a biotinylated ,fluorescent‐labelled ,lipid‐coated , liquid perfluorocarbon emulsion . Physico‐chemical properties of the agent were measured ,including size distribution ,Zeta Potential ,concentration and so on . Ischemia‐reperfusion models were created in rats ,and then exposed to biotinylated anti‐MCP‐1 monoclonal antibody ,rhodamine avidin and biotinylated ,FITC‐labelled nanoparticles ,respectively . Echocardiography was taken before and after injection . Frozen sections of their hearts were observed under fluorescence microscope . Results T he particle diameter ,zeta potential and concentration of lipid nanoparticles were ( 172 .30 ± 52 .06) nm ,( －33 .10 ± 6 .50) mV and ( 2 .28 ± 0 .46 ) × 1011/ml ,respectively . From the short‐axis view ,the myocardium under endocardium of anterior wall was enhanced obviously . While myocardium of other walls were still . T he lipid nanoparticles located in the myocardium of anterior wall and gave out bright green and red fluorescence under fluorescence microscope ,w hile neither lipid nanoparticles nor fluorescence were found in other sites of ventricular myocardium . Conclusions The viable myocardium can be targeted and acoustically enhanced by the self‐made nano‐scale ultrasound contrast agent . T his new agent has potential to improve sensitivity and specificity for noninvasive identifying viable myocardium .

Fabrication of open porous microtissue based on micro-stencil array chip / 中华整形外科杂志

Objective@#To explore an open porous microtissue which is suitable for the growth of bone marrow mesenchymal stem cells (BMSCs) , and the optimal size of this microtissue.@*Methods@#Polymethyl methacrylate (PMMA) micro-stencil array chips with pore diameters of 1500 μm and 800 μm were constructed by laser drilling technique. The gelatin solution was chemically crosslinked and uniformly spread on the micro-stencil array chips. After freeze-drying for 16 h, the microgels in the chip were ejected by the ejector-pin array. SEM images were taken to observe the microstructure of microgels. Pore size and porosity of two kinds of microgels were calculated using Image J Pro software. The P3 BMSCs were seeded on microgels at the concentration of 5×106/ml. After 7 days culture in vitro, the microtissues were obtained. Thereafter, the viability of seeded BMSCs was assessed using live/dead cell assay. The growth of the cells on the microtissues and the secretion of ECM were observed by scanning electron microscopy (SEM).@*Results@#The SEM images showed that the open porous structured microgels were successfully fabricated using the micro-stencil array chip. The pore size and porosity of 1 500 μm microgels were about 1.5 times that of the 800 μm microgels. After 7 days culture in vitro, the number of cells 1 500 μm microgels were more compared to 800 μm microgels, in the live/dead cell assay. Fluorescent quantitative analyses showed that the viability of BMSCs on 1 500 μm microtissues was 4 times that of 800 μm microtissues. SEM images showed that the cells on 1 500 μm microtissues grew well and surrounded by ECM.@*Conclusions@#The micro-stencil array chips were successfully used to construct microgels with open porous, which could be further used to fabricate open porous microtissues. The 1 500 μm microtissues were more favorable for BMSCs, compared to 800 μm microtissues.

Analysis of the correlation between the extreme temperature of incision surface and non-union of incision in internal fixation of calcaneal fracture / 中国基层医药

Objective@#To explore the correlation between the extreme temperature of incision surface and non-union of incision in internal fixation of calcaneal fracture.@*Methods@#From January 2016 to December 2017, 142 patients with closed unilateral calcaneal fracture treated with open reduction and internal fixation in the Integrated Chinese and Western Medicine Hospital of Wenzhou were enrolled.The incision temperature was collected by infrared radiation thermometers intermittently for 4-5 times, and the extreme values were used for further analysis.The primary endpoint of the study was the incision healing 14 days after surgeries.The patients′ age, gender, inflammatory indicators and body mass index (BMI) were evaluated, as well as related factors such as diabetes and arteriosclerosis.@*Results@#All patients completed the follow-up observation 14 days after operation, and finally achieved the wound healing in 105 cases, and the wound non-healing in 37 cases.The average extreme value of low temperature was (34.91±0.47)℃ in the union group and (34.13±0.60)℃ in the disunion group, and the average extreme value of high temperature was (36.61±0.55)℃ in the union group and (36.90±0.52)℃ in the disunion group, there were statistically significant differences between the two groups(t=7.152, -2.762, P=0.000, 0.007). Logistic regression analysis results showed that age[β(regression coefficient)=-0.111], extreme high(β=2.728) or very low (β=-3.854), arteriosclerosis(β=4.597), history of diabetes(β=2.234) were risk factors for incision nonunion(P=0.046, 0.003, 0.001, 0.002, 0.024).@*Conclusion@#Extreme incision temperature is observed to be associated with an increased risk of operative incision disunion after open reduction and internal fixation for calcaneal fractures.

The effects of Xiaotan Huayu Liqiao formula on mesenteric artery function in rats exposed to chronic intermittent hypoxia / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of Xiaotan Huayu Liqiao formula (the Chinese Medicine) on mesenteric artery function in rats exposed to chronic intermittent hypoxia (CIH), and to explore the related mechanism.@*METHODS@#Forty-eight male SD rats were randomly divided into four groups as Normoxia, CIH, Formula+CIH and formula group. Rats were exposed to normoxia in the Normoxia and Formula group, or intermittent hypoxia in CIH or Formula+CIH group. Xiaotan Huayu Liqiao formula was given at 24g/kg by intragastric administration before intermittent hypoxia exposure. The pathological changes of mesenteric artery were determined by HE staining, and the relaxation of mesenteric artery (induced by acetylcholine(ACh) and L-arginine(L-Arg)) was recorded by microvascular ring technique. Serums of all rats were collected (0 d and 21 d) and the content of NO was detected by ELISA. The levels of endothelial nitric oxide synthase (eNOS) and p-eNOS were measured by Western blot.@*RESULTS@#Compared with Normoxia group, the mesenteric arterial endothelial injury and media thickening were observed and the relaxation of mesenteric artery was significantly reduced in rats exposed to CIH. The level of NO in serum and the ratio of p-eNOS/eNOS were also decreased in the CIH group. Xiaotan Huayu Liqiao formula administration improved the pathologic changes and dilatation function of mesenteric artery, increased the levels of NO and p-eNOS. Compared with Normoxia group,all the results were not observed significant difference in Formula group.@*CONCLUSION@#Xiaotan Huayu Liqiao formula increased the bioavailability of NO, and ameliorated the CIH induced mesenteric artery function injury.

Study on cardiovascular injury of anesthesia rat induced by chronic intermittent hypoxia / 局解手术学杂志

Objective To investigate the effect of chronic intermittent hypoxia(CIH)on the cardiovascular system in anesthetized rats. Methods A totally of 72 male SD rats were randomly divided into three groups(n=24),namely the ormoxia control group(control group), the normoxia anesthesia group(model group)and the chronic intermittent hypoxia group(CIH group).Rats of the control group breathe nor-mally.The model group was given intraperitoneal injection of 10%hydration with 0.3 mL/kg,and the CIH group was given chronic intermit-tent hypoxic stimulation with 8 h/d in addtion to the model group.The difference of ultrasonic echocardiography data,blood pressure,endothe-lin type-1,and endothelial nitric oxide synthase(eNOS)in rats of the three groups were compared.After 28 days,these rats were sacrificed to observe the changes of myocardial cell structure.Results In the control group,the myocardial morphology was normal,the cells arranged e-venly,and there was no swelling and inflammation.In the model group,the myocardial cells were evenly arranged without hypertrophy and in-flammatory changes.In the CIH group,the myocardial cells in the hypoxic group were not evenly arranged,and hypertrophy,swelling,deform-ation,hyperchromia,and obvious inflammatory changes of the myocardial tissue were observed.In the control group,myocardial cell nucleus and cytoplasm were uniformly arranged,and there was no obvious changes in the model group.On the contrary,myocardial cell morphology changed obviously in the CIH group,with the cell morphology and size of the inhomogeneity increased, and the color of the apoptosis cells changes from light to dark.The tail artery systolic pressure of rats in CIH group was significantly higher than that of the control group and the model group,and the LVEF of CIH group was significantly lower than that of the other two groups(P<0.05).Ultrasound detection value sug-gests that the LVID and left ventricular of rats in the CIH group were slightly larger,and the diastolic function was normal.The LVDs of the model group and the CIH group were both higher than that of the control group with statistically significant difference(P<0.05).The RBC, HCT,dp/dtmax,and -dp/dtmax in the CIH group were significantly higher than those of the control group and the model group(P<0.05). Serum levels of endothelin-1 in CIH group were significantly higher than that of control group and model group,while the summation of serum NO2-/NO3-and eNOS in CIH group were significantly lower than those of the control group and the model group(P<0.05).Conclusion Chronic intermittent hypoxia can cause cardiac dysfunction in anesthetized rats,which may lead to the imbalance of serum endothelin-1 and NO levels,leading to endothelial dysfunction and myocardial injury.

The role of adventitia in hypoxic vascular remodeling / 生理学报

As an important site for the production, storage and release of key regulators for vascular function, the vascular adventitia is thought to be a damage sensing tissue in the vascular wall under certain conditions. The adventitial cells are usually the first ones to respond to vascular stress or injury, and consequently affect the structure and function of blood vessel wall. Growing lines of evidence have shown that the vascular adventitia exhibits the earliest and most prominent changes in vascular remodeling due to hypoxia and related pulmonary hypertension and atherosclerosis. In particular, fibroblasts play an important role in the adaptation and regulation to local microenvironmental changes. This review focuses on the role of vascular adventitia in hypoxia-induced vascular remodeling and the underlying molecular mechanisms.

Bosentan ameliorates hypertension in rats exposed to chronic intermittent hypoxia through inhibiting renal sympathetic nerve activity / 生理学报

The purpose of this study is to investigate the effect of the oral endothelin antagonist Bosentan on blood pressure and renal sympathetic nerve activity (RSNA) in rats exposed to chronic intermittent hypoxia (CIH), and to explore the sympathoexcitation mechanism of endothelin-1 (ET-1) in CIH-induced hypertension. Twenty-four male SD rats were randomly divided into normoxia, CIH and Bosentan groups. Rats in the normoxia group were exposed to normoxic environment, and rats in CIH or Bosentan group were exposed to intermittent hypoxia for 3 weeks. Bosentan was given at 50 mg/kg by intragastric administration before intermittent hypoxia exposure in Bosentan group. Systolic blood pressure (SBP) was measured by BP-2000, and the change of RSNA to sodium nitroprusside (SNP) or phenylephrine (PE) was recorded by PowerLab signal acquisition system. Serums of all rats were collected and the contents of ET-1 and norepinephrine (NE) were measured by ELISA. Results showed that blood pressure was gradually increased following CIH exposure compared with the normoxia group during the 3 weeks (P < 0.01, P < 0.01, P < 0.001). The basal RSNA was increased and baroreflex sensitivity was decreased in rats exposed to CIH. Furthermore, the blood pressure was positively correlated with the level of ET-1 in serum in rats exposed to CIH (r = 0.833, P = 0.01). Bosentan administration significantly decreased SBP and basal RSNA, increased the baroreflex sensitivity, and decreased serum NE level in rats exposed to CIH. These results suggest that ET-1 is related with blood pressure elevation in rats exposed to CIH, and Bosentan reverses CIH-induced hypertension by decreasing RSNA.