Objective: To explore the effect of HIF-1α on osteogenic-angiogenic coupling response in bone mesenchymal stem cells (BMSCs) and provide new concepts for engineered bone tissue in vitro. Methods: With the approval of the hospital’s experimental animal ethics committee, BMSCs were harvested from Wistar rats. The lentivirus carrying hypoxia-inducible factor-1α (HIF-1α) and empty lentivirus were stably transfected into the third generations of BMSCs to form LV-HIF-1α-BMSCs and LV-BMSCs. Meanwhile, BMSCs without transfection of lentivirus were used as a blank control. Then, the effect of HIF-1α transfection was verified by qPCR and Western Blot. LV-HIF-1α-BMSCs were induced to differentiate into endothelium-like cells (iECs). The morphology was observed by optical microscopy, the differentiation rate was detected by cellular flow CD31, and the Transwell test was used to detect the migration ability. At the same time, LV-HIF-1α-BMSCs and LV-BMSCs were continuously cultured to form osteogenic cell sheets (OCTs), which were stained by alkaline phosphatase on day 14 and alizarin red staining on day 21, and counted for mineralization capacity. Finally, iECs were implanted into OCTs to form prevascularized osteogenic cell sheets (P-OCTs), immunofluorescence CD31 was performed to detect the formation of vascular networks, and the results were recorded on days 1, 3, 7, and 14. Meanwhile, osteopontin (OPN) and osteocalcin (OCN) were detected by western blot to verify their ability for osteogenic differentiation on days 1, 7, and 14. Results: The optimal multiplicity of infection (MOI) for lentiviral transfection was 30, and the transfection efficiency was >80%. The results of qPCR and western blot showed that compared with the LV-BMSCs group and BMSCs group, the LV-HIF-1α-BMSCs group had stable and high expressions of HIF-1α (P<0.05). LV-HIF-1α-BMSCs showed an enhanced ability to differentiate into endothelial cells, with a differentiation rate as high as 91.81%. Transwell assay verified that HIF-1α could recruit iECs in vitro. Alkaline phosphatase staining and alizarin red staining confirmed that OCTs formed by LV-HIF-1α-BMSCs had a statistically significant osteogenic differentiation ability compared with LV -BMSCs control group (P<0.05). When iECs were implanted into the LV-HIF-1α-BMSCs group OCTs to form P-OCTs, iECs substantially proliferated and rapidly fused, and formation of the progressive lumen was revealed by immunofluorescent CD31 staining. The expressions of OPN and OCN were significantly enhanced compared with those of the LV-BMSCs control group; OCN was the highest on day 7, and OPN was the highest on day 1 (P<0.05). Conclusion BMSCs transfected by HIF-1α have good osteogenic-angiogenic effect after induction and differentiation, which provides experimental foundation for optimizing the construction of three-dimensional prevascularized bone tissue.

Objective To investigate the impact of digital subtraction angiography (DSA)-guided neurointerventional thrombus removal combined with intravenous thrombolysis by tirofiban on the rate of recanalization of blood vessels, endothelial function, hemodynamics, and the degree of neurological deficit in patients with acute cerebral infarction. Methods Eighty patients with acute cerebral infarction were selected as study subjects and randomly divided into observation group and control group using the random number table method, with 40 patients in each group. The control group was treated with DSA-guided neurointerventional thrombus removal, and the observation group was treated with intravenous thrombolysis with tirofiban on the basis of the control group. The rates of recanalization of blood vessels, endothelial function index levels, hemodynamic index levels, and the degree of neurological deficit were compared between the two groups. Results The rates of recanalization of blood vessels in the observation group and control group were 90.00%(36/40) and 65.00%(26/40), respectively, with a significant difference (P < 0.05). After treatment, the serum endothelin-1 (ET-1) levels in both groups were lower than those before treatment, and the nitric oxide (NO) levels were higher than those before treatment, and the ET-1 level in the observation group was lower than that in the control group, and the NO level was higher than that in the control group (P < 0.05). After treatment, the carotid extracranial resistance and characteristic impedance in both groups were lower than those before treatment, and the mean blood flow and mean blood flow velocity were higher than those before treatment, and the carotid extracranial resistance and characteristic impedance in the observation group were lower than those in the control group, and the mean blood flow and mean blood flow velocity were higher than those in the control group (P < 0.05). After treatment, the National Institutes of Health Stroke Scale (NIHSS) scores in both groups were lower than those before treatment, and the NIHSS score in the observation group was lower than that in the control group (P < 0.05). Conclusion DSA-guided neurointerventional thrombus removal combined with intravenous thrombolysis by tirofiban can effectively increase the rate of recanalization of blood vessels in patients with acute cerebral infarction, improve endothelial function, alleviate neurological deficit symptoms, increase blood flow velocity and blood flow at ischemic lesion sites, and reduce carotid extracranial resistance and characteristic impedance.

Objective To analyze the epidemiological characteristics of an outbreak of carbapenem resistant Klebsiella pneumoniae(CRKP)hospital infection in the neonatal intensive care unit(NICU),and to explore possible causes of the outbreak.Methods Case analysis,epidemiological studies and environmental detection methods were used to investigate the detection of CRKP in the NICU of a hospital from March 11,2022 to May 3,2022,and the causes and corresponding prevention and control measures were analyzed.Results This CRKP outbreak in the NICU occurred in two phases.The first phase was from March 11,2022,to March 30,2022,during which 3 people contracted CRKP hospital infections.The second phase was from April 13,2022,to May 3,2022,during which 4 people contracted CRKP hospital infections.The source of the hospital infection outbreak was patient No.1.All 13 CRKP cases showed sensitivity only to amikacin and tigecycline,with intermediate sensitivity to levofloxacin,and high resistance to other antibiotics.All 13 CRKP cases carried resistant genes for metalloenzymes.After the outbreak,the ward environment was sampled three times,with 35 samples taken in total,and 14 of them tested positive for CRKP.After strict environmental cleaning,disinfection,and multiple control measures,no new hospital infection cases similar to this outbreak were detected after May 13,2022.Conclusion The outbreak of NICU hospital infections may be due to the contamination of the ward environment,medical equipment,and inadequate hand hygiene of medical staff,leading to its spread.Therefore,strict cleaning and disinfection of the environmental surfaces,strengthening hand hygiene of medical staff,and implementing multiple measures can effectively control hospital infections of CRKP.

Objective:This multicenter clinical evaluation analyzed the clinical performance of five fast nucleic acid detection systems for 2019-nCoV.Methods:Clinical performance of the five fast nucleic acid detection reagents approved in China was evaluated in the present study. Fifty-seven throat swabs samples from COVID-19 patients and fifteen throat swabs samples from healthy people were collected from the First Affiliated Hospital of Zhejiang University school of Medicine, Tongji Hospital of Tongji Medical College of HUST, and National Institute of Viral Disease Control and Prevention of CDC to evaluate the positive coincidence rate, negative coincidence rate, total coincidence rate, the detection time and retest rate as well as the relation between positive intensity and positive coincidence rate of the five fast nucleic acid detection systems in November 2020.Results:The positive coincidence rates of the five kits were 92.59% (50/54), 83.64% (46/55), 98.25% (56/57), 94.44% (51/54) and 98.18% (54/55); and the negative coincidence rates were 93.33% (14/15), 93.33% (14/15), 86.67% (13/15), 100% (14/14) and 93.33% (14/15); and the total coincidence rates were 92.75% (64/69), 85.71% (60/70), 95.83% (69/72), 94.20% (65/69) and 97.14% (68/70), respectively. The positive coincidence rate of the five kits reached 100% for the strong-positive (90/90) and medium-positive samples (84/84), but only 82.18% (83/101) for weak-positive samples (cycle threshold value>33), and the retest rate of two kits were 15.28% (11/72) and 12.50% (9/72), which were both higher than 10%. Total time from sample extraction to amplification was between 32.33-65.33 minutes for these five kits.Conclusion:The five fast nucleic acid detection reagents have good performance and can be used as a supplement to routine nucleic acid detection reagents.

Background: Patients with acute gouty arthritis experienc severe pain and often have concomitant limited joint movement. High-dose intake of anti-inflammatory analgesics often results in adverse reactions, such as nausea, vomiting, and diarrhea, and might increase the risk of gastrointestinal bleeding. In order to reduce the adverse reactions by oral intake, local use of externally useddrugs with the same active ingredients as the oral intake is an option in clinical practice. However, externally useddrugs take effect slowly and efficacy to reduce painful feelingis unsatisfactory. Therefore, it is demanding to develop a safe and effective treatment plan. In this regard, researchers have put up a hypothesis. The skin of the human body has a dense stratum corneum, which is difficult for externally used drugs to penetrate effectively; if the keratin barrier of the skin around affected joints can be broken without further damaging the skin, externally applied drugs may safely and effectively alleviate the local symptoms of acute gouty arthritis. The nano plum blossom needle is made of a modern new material. The needle body is thin and the diameter of the needle tip is only 300 nm. The needle is used clinically to break through the keratin barrier of the human skin and promote the absorption of external medicine. Therefore, this trial was designed to preliminarily evaluate the safety and effectiveness of using the nano plum blossom needle for the introduction of diclofenac diethylamine emulgel in the treatment of acute gouty arthritis. Methods: A randomized controlled trial including 83 patients with acute gouty arthritis was conducted, who was assigned to three groups, the intervention group, control group 1, and control group 2 in a ratio of 2:2:1, respectively. The nano plum blossom needle was used for the introduction of diclofenac diethylamine emulgel in the intervention group; the nano plum blossom needle was used as a placebo along with the local use of diclofenac diethylamine emulgel in control group 1; and the nano plum blossom needle was used alone in control group 2. The treatments were applied once a day until the pain was completely relieved and the course of treatment lasted up to 7 days. The primary outcome measurement was the visual analog scale for evaluating the degree of pain. The secondary outcome indicators included scoring of the symptoms and signs with comprehensive consideration of the joint skin color, local tenderness, and degree of joint motion, the time and dose of emergency medication usage during trial, and adverse events. Discussion: This trial couldprovide preliminary evidence for evidence-based practice of using nano plum blossom needle transdermal drug delivery technology for diclofenac diethylamine emulgel for the treatment of acute gouty arthritis, and provide a reference for sample size calculation and experimental design of future clinical trials verifying the effectiveness and safety of such a technical scheme in a larger target population. Registration information This study has been registered in the China Clinical Trial Registry Centerwith the registration code of ChiCTR-IOR-17012154.

Qinling is a treasury of medicine with abundant resources of herbs. Due to the goal of enriching teaching contents, quality of teaching and creating a Chinese materia medica teaching system. Based on the Qinling plant gene database, our education team presents a series of innovative thoughts, such as multidisciplinary contact, inheriting and carrying forward the folk medicine, attaching importance to the practical application of herbs in Qinling and theory with practice to enrich teaching contentsand creating Shanxi characteristic teaching system of Chinese materia madica.That makes a contribution to raise the level of teaching in modern traditional Chinese medicine and promote the development of medicine in Shannxi.

OBJECTIVE:To formulate the quality standard of Cnidium monnieri liniment (called "liniment" for short)preliminarily. METHODS:The property of liniment was observed. TLC was used for qualitative identification of osthole in liniment;relative density,alcohol content and pH value were also determined. The osthole and imperatorin in liniment were analyzed qualitatively by HPLC. RESULTS:3 batch of samples were reddish brown liquid and fragrant smelling. In TLC of test sample,the same color fluorescent spots were found in corresponding position as chromatogram of control sample. The linear range of osthole and imperatorin were 0.101 2-0.910 8(r=0.999 6)and 0.006 2-0.124 4 mg/L(r=0.999 6). RSDs of precision tests were 1.38% and 0.79%(n=6). RSDs of stability tests were 0.33% and 0.41%(n=6). RSDs of reproducibility tests were 0.83% and 1.98%(n=6),respectively. Average recoveries rate were 98.73%(RSD=1.29%,n=6)and 99.25%(RSD=1.22%,n=6).Results of content determination showed that the content of osthole and imperatorin were 2.20-2.35 mg/mL and 0.310-0.340 mg/mL,respectively. CONCLUSIONS:Established method is simple,rapid,accurate and can be used for quality control of C. monnieri liniment.

Objective:To study the valuation and uncertainty evaluation of the purity determination of aminopyrine by two different principle methods. Methods:According to Technical Norm of Primary Reference Material and Technical Norm of Measurement,HPLC and acid-base titration were selected for studying the valuation of the purity determination of aminopyrine, and the uncertainty evaluation of the two different principle methods was systematically evaluated. Results:By using the two different principle methods,the standard value and uncertainty of aminopyrine content was 99.66% ± 0.08%(k = 2,P = 0.95). Conclusion:The valuation and uncertainty evaluation of the purity determination of aminopyrine by using HPLC and acid-base titration are accurate and reliable,which can avoid the defects by using single analysis method,and is helpful to improve the level of quality evaluation and control of aminopyrine. The study provides scientific basis for the development of aminopyrine purity reference materials.

Objective: To establish a GC method for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.Methods: The sample was dissolved in water, alkalified by sodium hydroxide solution and extracted by methylene chloride.An HP-5 gas chromatography column (50 m×0.32 mm, 1.05 μm) was used.The column temperature was programming increased, and the initial temperature maintained at 120 ℃ for 3 min, and then raised to 220 ℃ at a rate of 10℃·min-1 and maintained for 7 min.A hydrogen flame ionization detector (FID) was used and the split ratio was 1∶1.The inlet temperature was 230 ℃ and the detector temperature was 260 ℃.The injection volume was 1 μl and the carrier gas was nitrogen with high purity at a flow rate of 3.0 ml·min-1.Adamantane was used as the internal standard, and the internal standard method was used for the calculation.Results: The calibration curve was linear over the range of 0.05-1.0 mg·ml -1 (r=0.999 7).The detection limit and the limit of quantification was 1.1 ng and 3.3 ng, respectively.The average recovery was 100.2% (RSD =0.73%, n=9).Conclusion: The method has the advantages of simple operation, small extraction process toxicity, little environmental pollution, high accuracy and high specificity, and can be used for the determination of content and content uniformity of memantine hydrochloride dispersible tablets.