Objective To investigate the effect of dexamethasone on the toxicity of bupivacaine in murine neurons.Methods Murine neuroblastoma cell line N2a was obtained from ATCC cell bank (USA). The cells were cultured in 10% fetal cow serum/MEM culture medium and divided into 4 groups voup I control (Con); group II bupivacaine ( Bup); group Ⅲ dexamethasone (Dex) and group IV Dex + Bup. The culture medium contained bupivacaine 900 μmol/L in group Bup and dexamethasone 1 μmol/L in group Dex respectively. In group Dex + Bup ( IV ) Bup was added to the culture medium with a final concentration of 900 μmol/L at 12 h after pretreatment with Dex 1 μmol/L. The cells were inoculated in 24 well plates (0.5 ml in each well, 24 wells in each group) and 10 cm culture dishes (7 ml in each dish, 4 dishes in each group). The release rate of LDH was calculated and the morphology of the cells and nucleus condensation (by Hoechst 3334224 fluorescent staining) was detected at 9 h of incubation in 24 well plates. The mitochondrial transmembrane potential (by JC-1 assay) and phosphorylation of Akt and ERKs (by Western blot) were measured at 5 h of incubation in 24 well plates and in culture dishes respectively. ResultsBupivacaine caused severe damage to the N2a cells as evidenced by increase in LDH release and nucleus condensation (apoptosis), dephosphorylation of Akt and ERKs, decrease in mitochondrial transmembrane potential and severe morphological changes. Dexamethasone pretreatment significantly attenuated bupivacaine-induced neurotoxicity. Conclusion Dexamethasone can protect N2a cells from bupivacaine-induced neurotoxicity through stabilization of mitochondrial transmembrane potential and inhibition of dephosphorylation of Akt and ERKs.

Objective To retrospectively analyzed the current application of warfarin in hospitalized patients with non-valvular atrial fibrillationand ( NVAF), explore the key role of clinical pharmacists in warfarin medication. Methods A retrospective survey of anticoagulant therapy for 179 hospitalized patients with non-valvular atrial fibrillation in Renming Hospotal of Wuhan University from January to December 2013 was retrived,including the usage of warfarin for NVAF and new-onset atrial fibrillation,dosage,international normalized ratio(INR),hemorrhage event and so on.The simple factor like the age,complicated chronic diseases and previous cerebrovascular events on the use of warfarin was explored. Results The total response rate to anticoagulants was 85.6% for patients with high risk of stroke(27.3% with warfarin and 58.3% with antiplatelet therapy),who are recommended to use warfarin,patient were treated with anti-thrombotic therapy.The total of 19.1% of the patients with new-onset atrial fibrillation used warfarin as therapy.The whole monitoring rate of INR was 89.8%,and the good control rate was 11.9%. Univariate analysis showed that some high risk factors such as age and high blood pressure affected the usage of warfarin. Conclusion The anti-thrombotic therapy for NVAF patients in the hospital is good,but usage of warfarin for those with new-onset atrial fibrillation is low,which couldn't reach the INR standard. More attention should be taken by the clinic pharmacists in effective managing the use of anticoagulant to build a safe,economic and effective medication system for warfarin application.

Objective To investigate the existence of genetic divergence of sympatric populations of Anopheles sinensis of different feeding preferences based on the rDNA-ITS2 sequence differences. Methods A large number of wild anopheles popu-lations were trapped all night by man-baited net and calf-baited net that had been set up between high-density natural villages of An. sinensis populations and vector-breeding sites,from which two groups of An. sinensis were separated by morphological iden-tification and brought back to the lab for conventional breeding. A large closed greenhouse which temperature and humidity was appropriate was selected as research settings of mark-release-recapture methods by female mosquitoes ,in the center of which above An. sinensis populations baited by man and calf and respectively correspondingly marked by red and yellow phosphors were released in together,in each side of which An. sinensis were recaptured simultaneously by man-baited net and calf-baited net. An. sinensis populations trapped by man twice were brought back to the lab and bred with man-blood,correspondingly ones trapped by calf with calf-blood. Man-preferring and calf-preferring strains were screened respectively from An. sinensis which had been baited by man and calf by the mark-release-recapture methods after parent and F1 mosquitoes,and sequencing and aligning of both rDNA-ITS2 were conducted via PCR amplification. Results The recapture ratios of wild parental mosquitoes An. sinensis of man-preferring group by man-baited net and calf-baited net were 54.07%(339/627)and 45.93%(288/627)re-spectively,and ones of calf-preferring group by man-baited net and calf-baited net were 58.01%(409/705)and 41.99%(296/705)respectively. Two groups of parental mosquitoes trended towards selecting the original blood hosts in host-seeking prefer-ence(χ2=19.42,P<0.01). The recapture ratios of F1 mosquitoes An. sinensis of man-preferring group by man-baited net and calf-baited net were 63.43%(765/1 206)and 36.57%(441/1 206),and ones of calf-preferring group by man-baited net and calf-baited net were 68.22%(1 039/1 523)and 31.78%(484/1 523). Two groups of F1 mosquitoes had more significant characteris-tics of selecting the original blood hosts in host-seeking preference(χ2=271.69,P<0.01)and showed the genetic differentia-tion phenomenon,but the results of sequencing and aligning of the rDNA-ITS2 via PCR amplification showed no difference in base sequence between the two strains and both were 469 bp. Conclusions The genetic divergence based on the rDNA-ITS2 se-quence does not happen in An. sinensis sympatric populations of different feeding preferences.

Objective To investigate the expressions of fatty acid-binding protein 5 (FABP5)and dihydroli-poamide dehydrogenase(DLD)in skin lesions of symmetrical acrokeratoderma(SAK), and to explore their significance. Methods Biopsy specimens were obtained from skin lesions on the wrists and perilesional skin of 9 patients with SAK, and from normal skin in the wrists of 9 healthy volunteers (control group). Reverse transcription PCR (RT-PCR)and immunohistochemical staining were performed to measure the expressions of FABP5 and DLD in these specimens. Results RT-PCR showed no significant differences in the mRNA expressions of FABP5 or DLD between lesional, perilesional and normal control skin specimens(both P > 0.05). Immunohistochemically, there was a significant increase in the extent and intensity of staining for FABP5 in SAK lesions. Concretely speaking, FABP5 was strongly expressed in the stratum corneum, granular and spinous layers in SAK lesions, but weakly expressed in the stratum corneum, granular and spinous layers in perilesional skin, and only in spinous and basal layers in normal control skin. The expression of DLD decreased in SAK lesions, and was observed only in the stratum corneum and spinous layer in a few cases of SAK. However, the full-thickness epidermis stained positive for DLD in perilesional skin, with the nuclei and cytoplasm both stained deep brown. Conclusion The overexpression of FABP5 in SAK lesions may participate in dysdifferentiation of keratinocytes, while the down-regulation of DLD expression suggests an imbalance in energy metabolism.

Objective To investigate the training expectations and training strategies of orthopedic clinical specialist nurses (OCSN).Methods Totally 5 020 orthopedic nurses from 342 hospitals from 30 provinces and cities were selected bv convenience sampling.Participants were investigated hy WeChat platform with self-designed questionnaire.Results A total of 4 982 effective questionnaires were collected.Among investigated nurses,13.7％ of them received orthopedic specialist nurses training;91.4％ believed that training should be carried out.Qualifications should be:college degree and above,senior nurse and above,at least 3 ～5 years of nursing experience,1～3 years of nursing experience in orthopedic department.For curriculum,public courses expected to be arranged were communieation skills,nursing teaching,nursing management,and nursing research;expected professional courses were functional exercises,pain management,extremity injury nursing,common treatment techniques in orthopedic department,traction nursing,and position nursing.It was suggested that training was divided into subspecialties and off-duty;classroom teaching,teaching rounds,case discussion and experience exchange were expected training methods;"theory-practice-theory-practice" was most preferable traiuing mode.The preferred teachers were orthopedic doctors,orthopedic head nurses,orthopedic specialist nurses,and senior orthopedic nurses.The length of training was expected to be three months,time for clinical practice should be greater than or equal to theoretical teaching,and the practice bases should be tertiary hospitals.Evaluation should be performed before completion,recertification could be later than completion,and the interval time of recertification should be within 5 years.Conclusion Training needs of orthopedic nurses for OCSN are strong.Training expectations(contents,length,modes and teachers) of orthopedic nurses should be considered when designing systematic training program on OCSN.

Objective To investigate the correlation between carotid intima-media thickness (CIMT) and left atrial and left ventricular enlargement in patients with acute cerebral infarction. Methods A total of 224 patients with acute cerebral infarction were included. Based on the thickness of CIMT, it was divided into three groups which were normal CIMT group, thickening CIMT group, and carotid plaque (CP) group, with 57, 97, and 70 patients included respectively. Clinical data were collected, and carotid artery color Doppler ultrasound, cardiac color Doppler ultrasound and other examinations were determined to carry out relevant statistical analysis.Results The left anterior-posterior diameter (LAD) , left atrial diameter index (LADI) , left ventricular end-diastolic septal thickness (IVSD) , and left ventricular mass index (LVMI) in the CP group were all higher than those in the normal CIMT group and thickening CIMT group (P < 0.05). The percentage of the left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF) took in the thickening CIMT group were both higher than those in the CP group (P < 0.05). Multi-factor logistic regression analysis indicated that there were statistically significant differences in age, homocysteine and left LVMI (P < 0.05). In the Pearson correlation analysis, CIMT and LADI were positively correlated (r= 0.184, P < 0.01) , and there was a positive correlation between CIMT and LVMI (r = 0.236, P < 0.01). Conclusions Left ventricular enlargement is one of the highrisk factors for CIMT abnormalities in patients with acute cerebral infarction. Left atrial and left ventricular enlargement are closely correlated to the severity of CIMT in patients with acute cerebral infarction, indicating that abnormal CIMT in patients with acute cerebral infarction has a certain predictive effect on left atrial and left ventricular enlargement.

Objective To summarize the clinical characteristics of children with coronary artery aneurysms (CAA) and thrombosis in Kawasaki disease (KD),in order to explore the safe and effective thrombolytic therapy and its prognosis.Methods The clinic,treatment and follow-up data of 210 patients with KD between January 2006 and December 2016 were retrospectively reviewed in the Department of Pediatrics,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology.The clinic signs and laboratory data for CAA with thrombosis were analyzed.The characteristics of CAA were monitored by tltrasound.All KD patients with thrombus received intravenous antithrombotic therapy,including urokinase,heparin,and oral Warfarin,and anti-platelet treatment.The effectiveness of antithrombotic treatment was evaluated by measuring the ability to dissolve the thrombus.Results Fourteen cases in 210 patients with KD developed CAA,and had associated thrombosis.In these 14 patients,the largest diameter of CAA was 18.5 mm,and the average value was 12.6 mm.There was no special blood analysis in CAA with thrombus.Moreover,typical KD symptoms and acute myocardial infarction were not found in CAA with thrombosis.Thrombus occurred in giant aneurysms,and 2 patients had multiple thrombosis.After thrombolytic therapy,12 cases in the 14 patients had successful thrombolysis,2 patients had thrombus organization and coronary artery stenosis.Conclusions Neither clinical features nor laboratory data could reliably predict CAA associated thrombosis.Thrombus was easily formed in giant CAA.Frequent and periodly follow-up are important to detect thrombosis in KD patients with giant coronary artery.Therapy with adequate intravenous antithrombotic therapy and anti-platelet treatment earlier can effectively dissolve thrombus in KD patients,and avoid deterioration.

Objective:To study the clinical and molecular characteristics of a family with familial hypercholesterolemia (FH) with LDLRAP1 and ABCG8 gene abnormality.Methods:In September 2020, one case of FH was included in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine; peripheral venous blood samples of members of the family were collected to detect serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) indicators; use high-performance liquid chromatography to detect serum stigmasterol and sitosterol content; perform second-generation gene sequencing to detect gene mutations in probands and family members; use Pymol software to detect gene mutations point for pathogenicity analysis, and use Uniprot Modelling software to perform protein structure modeling.Results:The patient presented with anemia, multiple xanthomas and early-onset acute coronary syndrome. The coronary angiography showed severe coronary artery lesions; abdominal ultrasound showed splenomegaly; blood smear showed shaped erythrocytes and large platelets. The level of serum TC, LDL-C, stigmasterol and sitosterol was 8.54 mmol/L (2.3-5.7 mmol/L), 4.84 mmol/L (range of normal value 1.3-4.3 mmol/L), 44 μmol/L (1.0-10 μmol/L), 28 μmol/L (1.0-15 μmol/L), respectively; LDLRAP1 gene mutation was found: exon4 c.415C>T:p.Q139X; the truncated protein formed by this homozygous mutation lost multiple stable protein structure regions, which can not have a normal function. At the same time, ABCG8 gene mutations were also found: exon13 c.1895T>C (p.V632A) and exon8 c.1199C>A:p.T400K . Two cases of family members had a mild increase in HDL-C (Ⅱ5: 2.33 mmol/L, Ⅱ6∶2.96 mmol/L), 3 cases carrying the ABCG8 gene mutations had a slight increase in stigmasterol (Ⅱ8: 23 μmol/L, Ⅱ7: 24 μmol/L, Ⅰ2: 18 μmol/L) and sitosterol (Ⅱ8: 41 μmol/L, Ⅱ7: 33 μmol/L, Ⅰ2: 45 μmol/L), suggesting that its association with the concentration of plant sterols. Conclusions:FH patients with LDLRAP1 and ABCG8 gene abnormalities may have abnormal plant sterol concentrations, and their clinical manifestations are more complicated. Therefore, family history, LDL-C, plant sterol levels, and genetic test results should be considered comprehensively.

Erythrocytes-camouflaged nanoparticles is an in vivo delivery system that uses erythrocytes or erythrocyte membrane nano vesicles as carriers for drugs, enzymes, peptides and antigens. This system has the advantages of good biocompatibility, long circulation cycle and efficient targeting. This review summarizes the type of carriers, their development history, the application of delivery strategies as well as their limitations and future challenges. Lastly, future directions and key issues in the development of this system are discussed.
Pharmaceutical Preparations ; Drug Delivery Systems ; Vaccines ; Erythrocytes ; Nanoparticles

The pathogenesis theory of"spleen deficiency and stasis toxin"in gastric cancer holds that spleen is the source of generation and transformation of qi and blood,that spleen deficiency is the internal basis of disease and throughout the disease.Stasis toxin is based on spleen deficiency,which is the fundamental pathogenesis of gastric cancer.In the pathological process of gastric cancer,a variety of metabolic substances in tumor cells and tumor microenvironment,mainly glucose metabolic reprogramming,undergo metabolic changes to reconstruct the phenotype and function of tumor-related macrophages,which is consistent with the pathogenesis theory of"spleen deficiency and stasis toxin".Therefore,this article focused on the reprogramming of glucose metabolism in tumor microenvironment to drive the phenotypic remodeling of tumor-related macrophages,explored the scientific connotation of the pathogenesis theory of"spleen deficiency and stasis toxin"of gastric cancer,and provided references for the theoretical and clinical research on the treatment of gastric cancer by TCM.