Objective:To develop an EGFR-targeting nanobody engineered exosome drug delivery system and evaluate its antitumor efficacy for colorectal cancer.Methods:The HEK293T cell line stably expressing the pan-cancer inhibitor miR-204-5p, previously established by our group, was selected as a tool cell line to prepare miR-204-5p-enriched exosomes. Using metabolic glycoengineering combined with bioorthogonal reaction strategy, these exosomes were modified with the EGFR-specific nanobody 7D12. Western blot, electron microscopy, and dynamic light scattering were used to characterize the engineered exosomes. The tumor target potential of engineered exosomes was evaluated using immunofluorescence and RT-qPCR. The in vitro anti-tumor activities of engineered exosomes were evaluated using cell growth curves, colony formation, Transwell, and apoptosis analyses. The in vivo anti-tumor activity and safety of engineered exosomes were evaluated using a nude mouse xenograft tumor model. Results:The particle size of 7D12-hExo was (116.8±36.8) nm, with a potential of around -10 mV, and there was no significant change compared with the unmodified hExo. Immunofluorescence assay showed that the fluorescence intensity of the hExo group, 7D12-hExo group, and 7D12+7D12-hExo group were 48.4±3.9, 141.0±6.6, and 38.7±3.2 in EGFR + HCT116 cells, respectively. Compared with the hExo group, the fluorescence intensity of HCT116 cells in the 7D12-hExo group was significantly enhanced ( P＜0.05). Compared with the 7D12-hExo group, the fluorescence intensity in HCT116 cells in the 7D12+7D12-hExo group was significantly decreased ( P＜0.05). However, there was no significant difference in the uptake of hExo and 7D12-hExo in EGFR - SW620 colorectal cancer cells. The number of cell clones, invasion, and migration of HCT116 cells in the hExo (204) group was 215.0±14.0, 862.3±61.4, and 1 197.0 ± 36.7, respectively, with an apoptosis rate of (14.1±1.4)%. The number of cell clones, invasion, and migration of HCT116 cells in the 7D12-hExo (204) group was (65.0±15.1), (232.0±27.9), (725.7±32.7), respectively, with an apoptosis rate of (29.3±1.0)%. The 7D12-hExo (204) significantly inhibited the proliferation, invasion, and migration ability of HCT116 cells ( P＜0.05), resulting in promoting the apoptosis of HCT116 cells ( P＜0.05). Nude mouse experiments showed that 7D12-hExo (204) significantly inhibited the growth of tumors transplanted with HCT116 cells, with the inhibition rate being 82.8%. However, there was no significant change in mouse weight, and H&E staining of major organs such as heart, liver, spleen, lung, and kidney did not show any abnormalities. Conclusion:Naturally miR-204-5p-loaded exosomes were successfully modified with nanobody 7D12, which can efficiently deliver miR-204-5p into EGFR + tumor cells, thereby exerting good anti-tumor therapeutic effects.

Objective:To develop an EGFR-targeting nanobody engineered exosome drug delivery system and evaluate its antitumor efficacy for colorectal cancer.Methods:The HEK293T cell line stably expressing the pan-cancer inhibitor miR-204-5p, previously established by our group, was selected as a tool cell line to prepare miR-204-5p-enriched exosomes. Using metabolic glycoengineering combined with bioorthogonal reaction strategy, these exosomes were modified with the EGFR-specific nanobody 7D12. Western blot, electron microscopy, and dynamic light scattering were used to characterize the engineered exosomes. The tumor target potential of engineered exosomes was evaluated using immunofluorescence and RT-qPCR. The in vitro anti-tumor activities of engineered exosomes were evaluated using cell growth curves, colony formation, Transwell, and apoptosis analyses. The in vivo anti-tumor activity and safety of engineered exosomes were evaluated using a nude mouse xenograft tumor model. Results:The particle size of 7D12-hExo was (116.8±36.8) nm, with a potential of around -10 mV, and there was no significant change compared with the unmodified hExo. Immunofluorescence assay showed that the fluorescence intensity of the hExo group, 7D12-hExo group, and 7D12+7D12-hExo group were 48.4±3.9, 141.0±6.6, and 38.7±3.2 in EGFR + HCT116 cells, respectively. Compared with the hExo group, the fluorescence intensity of HCT116 cells in the 7D12-hExo group was significantly enhanced ( P＜0.05). Compared with the 7D12-hExo group, the fluorescence intensity in HCT116 cells in the 7D12+7D12-hExo group was significantly decreased ( P＜0.05). However, there was no significant difference in the uptake of hExo and 7D12-hExo in EGFR - SW620 colorectal cancer cells. The number of cell clones, invasion, and migration of HCT116 cells in the hExo (204) group was 215.0±14.0, 862.3±61.4, and 1 197.0 ± 36.7, respectively, with an apoptosis rate of (14.1±1.4)%. The number of cell clones, invasion, and migration of HCT116 cells in the 7D12-hExo (204) group was (65.0±15.1), (232.0±27.9), (725.7±32.7), respectively, with an apoptosis rate of (29.3±1.0)%. The 7D12-hExo (204) significantly inhibited the proliferation, invasion, and migration ability of HCT116 cells ( P＜0.05), resulting in promoting the apoptosis of HCT116 cells ( P＜0.05). Nude mouse experiments showed that 7D12-hExo (204) significantly inhibited the growth of tumors transplanted with HCT116 cells, with the inhibition rate being 82.8%. However, there was no significant change in mouse weight, and H&E staining of major organs such as heart, liver, spleen, lung, and kidney did not show any abnormalities. Conclusion:Naturally miR-204-5p-loaded exosomes were successfully modified with nanobody 7D12, which can efficiently deliver miR-204-5p into EGFR + tumor cells, thereby exerting good anti-tumor therapeutic effects.

Objective:To compare the antiplatelet effects of vicagrel and clopidogrel in patients with different cytochrome P450 (CYP) 2C19 genotypes.Methods:This is a post-hoc analysis of a phase Ⅱ clinical trial of vicagrel, which included patients with coronary heart disease who underwent percutaneous coronary intervention from August 2018 to June 2019 in 18 centers. Patients were categorized based on the presence of CYP 2C19 *2 or *3 loss-of-function (LOF) alleles into LOF carrier group ( n=111) and non-LOF carrier group ( n=90). Each group included patients received vicagrel 5 mg, 6 mg, 7.5 mg, or clopidogrel 75 mg for 28 days per study protocol. P2Y 12 reaction units (PRU) were measured using VerifyNow at baseline, 6 to 8 hours after loading dose, 7 to 10 days after randomization, and 28 days after randomization and the percentage inhibition of platelet aggregation (%IPA) was calculated. The primary endpoint was %IPA on day 28. Within the patients from the General Hospital of Northern Theater Command, 8 to 12 patients in each study arms were enrolled in a prespecified pharmacokinetic sub-study, measuring the time to reach maximum plasma concentration (T max), peak plasma concentration (C max), and area under the plasma concentration-time curve (AUC). Results:Among 201 patients, the age was (58.8±8.5) years, and 139 (69.2%) were male. In non-LOF carriers, there was no significant differences in PRU values and %IPA between the vicagrel 5 mg, 6 mg, 7 mg, and clopidogrel groups at all time points (all P>0.05). In LOF carriers, %IPA was significantly higher in the vicagrel-treated groups than in the clopidogrel group at 6-8 hours after loading dose (22.9 (14.2, 31.5)% vs. 19.8 (11.0, 28.6)% vs. 29.5 (20.9, 38.0)% vs. 12.9 (3.9, 21.9)%, P=0.038) and 7-10 days after randomization (22.4 (14.2, 30.5)% vs. 34.4 (26.1, 42.6)% vs. 39.8 (31.8, 47.9)% vs. 24.7 (16.3, 33.2)%, P=0.001), with a trend towards higher %IPA in the vicagrel-treated groups at day 28 (30.4 (21.3, 39.6)% vs. 36.5 (27.2, 45.7)% vs. 40.8 (31.8, 49.8)% vs. 30.7(21.2, 40.2)%, P=0.056). Pharmacokinetic results of 35 patients showed that the C max and AUC of the active metabolite M15-2 of vicagrel was similar to that of clopidogrel in non-LOF carriers, but AUC between vicagrel 5 mg, 6 mg, 7 mg and clopidogrel were significantly different in LOF carriers ((5.6±0.6) h·μg -1·L -1 vs. (6.8±2.7) h·μg -1·L -1 vs. (9.2±3.3) h·μg -1·L -1 vs. (4.2±1.9) h·μg -1·ml -1, P=0.020). Conclusion:Vicagrel and clopidogrel have similar antiplatelet effects in non-LOF carriers, but vicagrel exhibits superior antiplatelet effects in LOF carriers.

Objective To investigate the effects of real-time feedback devices on chest compression quality test in non-medical staff during cardiopulmonary resuscitation (CPR) training.Methods A total of 120 volunteers were recruited and trained according to American Heart Association Guidelines Update for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care set in 2015.CPR performance with compression for six minutes was tested on a manikin.Volunteers were randomized into 3 groups.Group A was tested without any feedback.Group B was self-corrected in compression quality(include compression depth、rate and rebound of chest wall) using a real-time feedback device (Link CPR).Group C was guided with a metronome.All compression data were collected via WiFi signal and stored.Results Significantly better mean chest compression depth was achieved in group B than that in group A and C(5.38 ± 0.483 cm vs.4.42 ± 0.572cm and 4.25 ± 0.843 cm,P ＜ 0.05).Significantly better compression rate were observed in both group B and C than that in group A (113.4 ± 5.9 and 109.0 ± 6.8 compressions/min vs.129.6 ± 8.3 compressions/min,P ＜ 0.05).Significantly less rebounding were observed in both group B and C compared with group A (56.10 ± 32.3 and 68.30 ± 28.8 compressions vs.174.30 ± 38.8compressions,P ＜ 0.05).Pearson correlation analysis confirmed the compression rate was positively correlated with the numbers of rebounding (r=0.776,P＜0.01).Significant statistical difference in accuracy was observed among the groups (9.8％ vs.72.9％ vs.58.5％,P ＜ 0.05).Conclusions In CPR training test real-time feedback device contributes to the improvement of chest compression quality through self-adjustment of compression depth,rate and rebound.

Objective To observe the effects of electroacupuncture (EA) on c-Jun amino terminal kinase (JNK) signaling pathway and inflammatory cytokines interferon-γ (IFN-γ) in substantia nigra (SN) cells of rotenone-induced rats model of Parkinson's disease (PD),and explore the underlying mechanism of EA on PD.Methods A total of 32 male Sprague-Dawley mice were randomly and evenly divided into a normal group,a shamoperation group,a model group and an EA group.Model group and EA group were injected intradermally with rotenone (1 mg/kg,dissolved in DMSO and saline,concentration:O.25 mg/ml) on the nape of neck.Sham-operation group was injected the same dose of DMSO and saline.Normal group had no intervention.EA group was applied to Fengfu (DU16) and Taichong (LR3) acupoints after the establishment of PD model in rats.Behavioral assessment was conducted after the treatments,the rats were sacrificed for sampling substantia nigra tissue to detect the expressions of tyrosine hydroxylase (TH),p-c-Jun amino terminal kinase (p-c-Jun) and interferon-γ(IFN-γ) protein with Western blotting (WB).Results Model rats showed significant PD syndrome characteristics,comparing with normal group and sham group,the difference was not statistically significant (P ＞ 0.05).The results of open box test showed that the scores of model group rats decreased significantly in terms of the horizontal movement [(19.12 ±2.34) points] and vertical locomotor activity [(5.27 ± 1.04) points] when compared with normal group and sham group,the difference was statistically significant (P ＜ 0.05).After EA treatment,locomotor activity of rats increased significantly when compared with model group (P ＜ 0.05),however,the normal group and sham group was not statistically and significantly different in locomotor activity (P ＞ 0.05).Compared with normal group,the expression of TH protein in (0.183 ± 0.0213) reduced significantly and the expressions of p-c-Jun (0.388 ± 0.0283) and IFN-γ protein(0.453 ± 0.0332) increased significantly in model group,the difference was statistically significant (P ＜0.05).Compared with normal group,the expression of TH protein(0.324 ± 0.0538) reduced and the expressions of p-c-Jun(0.207 ± 0.0592) and IFN-γ protein (0.239 ± 0.0215) increased in EA group,the difference was not statistically significant (P ＞ 0.05).Compared with model group,the expression of TH protein increased significantly in EA group(P ＜ 0.05),the expressions of p-c-Jun and IFN-γ protein reduced significantly in EA group(P ＜ 0.05).Conclusion EA therapy may reduce the expression of IFN-γ protein in SN of PD rats model by regulating the expression of JNK/mitogen-activated protein kinase (MAPK) pathway,which may delay the process of PD.

BACKGROUND:The carriers made from biodegradable and biocompatible polymeric materials represent an exciting approach to increase the bioavailability and stability of oral y administered insulin by the chemical reaction or physical encapsulation of insulin.
OBJECTIVE:To mainly review the research progress of the material types, preparation methods, physicochemical characteristics, in vitro release kinetics, and bioavailability of polymeric materials adopted as oral insulin carriers.
METHODS:The first author searched PubMed, Elsevier and CNKI databases for articles (2002-01/2013-02) concerning the polymeric materials and oral insulin carriers with the key words of“polymeric biomaterials, oral insulin, carrier”in English and in Chinese.
RESULTS AND CONCLUSION:Currently, there are mainly two kinds of polymeric biomaterials used for oral insulin delivery systems, that is, natural polymeric biomaterials (such as chitosan and alginates) and synthetic polymeric biomaterials. The most commonly used synthetic polymeric materials for the preparation of these vehicles are polyesters, polyacrylates and their copolymers, which are wel known for their good biodegradability, biocompatibility, and physiological properties. Although researchers have tried to develop promptly oral insulin formulation using various technologies, the reports about clinical application or commercial success have not been seen because of several questions such as polymer material as a carrier, the lower bioavailability of insulin, the quality standards and stability of the formulation. Hence, future studies wil focus on the development of a new type of polymer-based material as carriers by choosing the new materials or modifying physical and chemical characteristics of existing polymers, to avoid gastrointestinal destruction of the insulin and increase bioavailability of insulin in the body, so as to obtain the good control ed release rate and effect.

Objective To explore changes of gray matter volume in patients with obsessivecompulsive disorder (OCD) in Chinese Han population using optimized voxel-based morphometry (VBM) ,and investigate its relationship with clinical symptoms. Methods Twenty patients with OCD and 20 age,sex and handedness matched healthy controls were scanned using 3D-T1 images on a 3.0 T MR system. The high resolution T1WI was preprocessed according to the optimized VBM protocol in Statistical Parametric Mapping (SPM5). Two-sample t test was performed to characterize the differences of the gray matter volume (GMV) between the OCD patients and healthy controls, and the correlations between the GMV and symptom severity and cumulative illness duration were examined using Pearson correlation in SPSS 16. 0, respectively.Results Compared to controls, OCD patients demonstrated increased GMV in left thalamus, right thalamus and left cerebellum after false discovery rate (FDR) correction. No areas of significantly decreased GMV was observed in OCD patients in relative to healthy controls. The mean eigenvalue ranged from 0. 5782 to 0. 889 representing the left thalamus volume of OCD patients was 0. 6813 ± 0. 0718, and that ranged from 0. 5546 to 0. 9062 was 0. 6869 ± 0. 0808 tor right thalamus. The mean eigenvalues were positively correlated in bilateral thalamus (r = 0. 94, P ＜ 0. 01). Conclusion Using optimized VBM, the current research indicates that the pathophysiology of OCD is associated with GMV abnormalities not only in corticostriato-thalamo-cortical (CSTC) circuit, but also in the cerebellum.

Objective To determine the prevalence of cervical type-specific human papillomavirus (HPV)infection as well as risk factors associated in Tibet Autonomous Region of China.Methods A cluster sampling study was performed in Lasa,Rikaze and Naqu of Tibet.An epidemiological questionnaire was applied and 3036 cervical specimens were obtained for liquid-based cytology and HPV DNA detection.Statistical analysis included Wald Chi-square and stepwise logistic regression model.Results The overall HPV prevalence of involved 3036 women was 9.19%(279/3036),of which 7.05%(214/3036)of the women were infected by high-risk types (including 14 sorts of types) and 2.14%(65/3036)by low-risk types(including 6 sorts of types).There were no significant differences of HPV prevalence between age groups(P=0.936),race(P=0.718)and areas(P=0.746),respectively.Twenty-one types of HPV were detected,of which HPV16(1.52%) was the most common type,followed by HPV33(1.42%).HPV58(1.22%),HPV52(1.15%),and HPV31(1.05%).HPV type distribution was varied by age.Of the 279 HPV infected women.14.3%(40/279)exhibited multiple HPV infections.Independent risk factors for HPV infection were smoking(P=0.027),number of sex partners(P=0.198)and early age of first intercourse(P=0.237).Conclusion The overall prevalence of HPV infection in Tibet Autonomous Region is lower than that in China or abroad,in which the most common genotype is HPV16 and the independent risk factors for HPV infection included early age of first intercourse,smoking,and number of Bex partners.

Objective To Evaluate the cognitive function in essential hypertensive patients complicated with metabolism diseases. Methods Eighty five essential hypertensive patients(EH)and other seventy four essential hypertensive patients complicated with metabolic disorder (EH+MD) were enrolled. Patients of EH+MD were categorized into glucose metabolism disorder (n=27), hyperlipidemia (n=15) and gluco-lipid metabolism disorder (n=22). Cognitive function was evaluated using "Clinical memory measuring scale" and "CISA intelligence scale". Results Direction and association memory, free imaging and portrait memory were better in EH than that in EH+MD (P

Objective To analysis the parameters of left ventricular hypertrophy(LVH) in patients with primary aldosteronism(PA).Methods Two hundred fifty patients were diagnosed as primary aldosteronsim in this study.One hundred fouty two PA patients undergone the adrenal venous sampling(AVS).Adenomas) APA) were diagnosed in 68 patients and idiopathic hyperaldosteronism(IHA) in 74 patients.During the same period, clinical characteristics and cardiovascular events of this group were compared with those of 246 patients with essential hypertension(EHT) randomly matched for age,gender,and systolic and diastolic blood pressure.Echocardiographic examinations and biochemical and hormonal assays were conducted in all subjects.Results 1) Left ventricular end-diastolic dimension(LVEDd) was increased in PA patients compared with values in EHT patients(PA;49.6?4.3 vs EHT:48.3?4.2 mm,P0.05);however,in grade 2 and 3 hypertension,the LVMI in patients with PA was increased compared with values in patients with EHT(P