ObjectiveTo investigate whether elemene（ELE） enhances the anti-glioma efficacy of cabazitaxel（CTX）， and prepare a double-targeted cationic liposome（LIP） co-loaded with ELE/CTX for the treatment of glioma， and to achieve the effect of increasing the efficacy and reducing the adverse reactions. Pharmacodynamic tests in vitro were performed to explore the advantages and mechanism of its preparation. MethodELE/CTX@LIP was prepared by high speed shear combined with probe ultrasound， the particle size and potential were characterized by nano-particle size potentiometer， and high performance liquid chromatography（HPLC） was used to determine the encapsulation efficiency and drug loading capacity of CTX/ELE. The cytotoxicity of ELE/CTX in vitro was detected by cell proliferation and activity assay（CCK8）. JMP Pro 16 software was used to optimize the process parameters of ELE/CTX@LIP based on encapsulation efficiency. The optimal cationic material type， content and ratio were screened by in vitro cytotoxicity and in vitro cell uptake， on this basis， the dual-targeted cationic liposome T7/arginine glycine aspartate tripeptide sequence（T7/cRGD）-ELE/CTX@CLIP was prepared， the stability of morphology and particle size were characterized， and the effect of T7/cRGD-ELE/CTX@CLIP on the apoptosis inducing ability and cell cycle regulation ability of glioma cells was analyzed by cell cycle and apoptosis. ResultELE/CTX showed stronger anti-glioma activity on C6 and RG2 cells. The results of in vitro cytotoxicity and in vitro cell uptake showed that the amount of cationic material was 0.10% of the total content. The optimum ratio of T7， cRGD and phospholipids was 1∶1∶50. T7/cRGD-ELE/CTX@CLIP［1，2-dilinoleyloxy-3-dimethylaminopropane（Dlin-MC3-DMA）］ and T7/cRGD-ELE/CTX@CLIP［1，2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol 2000（DMG-PEG2000）］ showed multi-level spherical nanostructures with particle sizes of 146.0， 111.3 nm， respectively， and were stable in serum. In vitro cytotoxicity results showed that T7/cRGD-ELE/CTX@CLIP had higher cytotoxicity to glioma cells than single-targeted liposomes or dual-targeted non-cationic liposomes. T7/crGD-ELE/CTX@CLIP affected the apoptosis and cycle of glioma cells， the results showed that ELE/CTX combined with liposomes could more effectively activate the apoptosis channel and inhibit the proliferation of glioma cells， and the use of T7/cRGD short peptide and cation modification enhanced the ability of apoptosis induction. ELE/CTX could effectively block glioma cell cycle at G₂/M phase， and the effect was enhanced after T7/cRGD targeted modification. ConclusionELE can enhance the anti-glioma effect of CTX. The preparation parameters of ELE/CTX@LIP are stable and feasible. Combined with the in vitro efficacy test， the anti-glioma mechanism of T7/cRGD-ELE/CTX@CLIP is preliminarily revealed.

Objective:To investigate the impact of geniposide(GE)on lung injury in rats with acute respiratory distress syn-drome(ARDS)by regulating AMP-activated protein kinase(AMPK)/silencing information regulator 1(SIRT1)/nuclear factor κB(NF-κB)signaling pathway.Methods:The ARDS rat model was established by tracheal instillation of lipopolysaccharide(LPS).Fifty rats after modeling were randomly group into ARDS group,GE low-dose(GE-L,12.5 mg/kg GE)group,GE medium-dose(GE-M,25 mg/kg GE)group,GE high-dose group(GE-H,50 mg/kg GE)group and GE-H+Compound C(AMPK inhibitor,50 mg/kg GE+250 μg/kg Compound C)group,another 10 normal rats were used as the control group.After the intervention,the bronchoalveolar la-vage fluid(BALF)and lung tissue of the rats in each group were taken out,respectively,and the ratio of lung wet to dry weight(W/D)was detected;ELISA was used to detect the levels of inflammatory factors IL-6,interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)in BALF;the positive expressions of vascular cell adhesion factor(VCAM-1)and vascular endothelial cell growth factor(VEGF)in lung tissue were detected by immunohistochemistry;HE staining was used to observe the pathological changes of lung tissue;Western blot was used to detect the expression levels of AMPK/SIRT1/NF-κB pathway proteins in lung tissue.Results:The levels of W/D,IFN-γ,IL-6,TNF-α,p-NF-κB p65/NF-κB p65 and VCAM-1 in ARDS group were significantly higher than those in control group,the expressions of p-AMPK/AMPK,SIRT1 and VEGF were significantly decreased(P<0.05);after different doses of GE treatment,the levels of W/D,IFN-γ,IL-6,TNF-α,and the expressions of p-NF-κB p65/NF-κB p65 and VCAM-1 were gradually decreased compared with those in ARDS group;the expressions of p-AMPK/AMPK,SIRT1 and VEGF increased gradually(P<0.05);Compound C reversed the protective effect of GE-H on ARDS rats(P<0.05).Conclusion:GE can improve lung injury in ARDS rats and reduce levels of inflammatory factors,which may be related to activation of AMPK/SIRT1/NF-κB signaling pathway.

Objective:To investigate the clinical characteristics of patients with keratoconus, and to explore the factors influencing keratoconus severity.Methods:A cross-sectional study was performed.A total of 908 patients (1 476 eyes) with primary keratoconus were enrolled in Henan Eye Hospital from January 2019 to December 2021.The medical history data of patients were collected by face-to-face questionnaire survey.Refractive parameters were measured by subjective optometry.Intraocular pressure (IOP) was measured by a non-contact tonometer, and corrected IOP was calculated by Dresden formula.Corneal topography parameters was obtained using Pentacam HR.The subgroup analysis of clinical characteristics of all patients was performed by age (<21 years, 21~<31 years, ≥31 years) and gender.Disease severity was graded based on steep keratometry (Ks), namely mild (Ks<48 D), moderate (48 D≤Ks<55 D) and severe (Ks≥55 D). The influencing factors of disease severity in keratoconus were analyzed by ordered Logistic regression.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[5]). All subjects or guardians were informed of the purpose and significance of the study and written informed consent was obtained.Results:Of the 908 patients, 622 were with bilateral keratoconus and 286 were with unilateral keratoconus.The median age of onset was 20(17, 26) years, and the median age of diagnosis was 21(18, 27) years.The ratio of males to females was 3.05∶1.There were 9.80%(89/908) of the patients having a history of allergy, 25.55%(232/908) having a history of other systemic diseases, and 1.98%(18/908) having a family history of keratoconus.Of the 1 476 affected eyes, 27.57%(407/1 476) were diagnosed as severe keratoconus, and 61.94%(568/917) had a history of eye rubbing.The medians of sphericity, cylindricity, IOP, corrected IOP, Ks, thinnest corneal thickness (TCT), anterior corneal surface elevation (AE) and posterior corneal surface elevation (PE) were -4.00(-7.00, -1.75)D, -3.50(-6.00, -1.50)D, 12.00(10.30, 13.80)mmHg, 15.40(13.60, 17.00)mmHg, 49.85(46.40, 54.90)D, 460.00(425.00, 490.00)μm, 21.00(13.00, 34.75)μm, 51.00(33.00, 75.00)μm, respectively.The spherical refraction, IOP and corrected IOP were lower and the cylindrical refraction was higher in patients at age <21 years than in patients at age 21~<31 years, and the TCT of patients at age <21 years was higher than that at age ≥31 years, and the differences were statistically significant (all at P<0.05). Compared with female patients, male patients had younger onset age, lower spherical refraction, IOP and corrected IOP, as well as higher cylindrical refraction, AE and PE, showing statistically significant differences (all at P<0.05). The spherical refraction and IOP of male patients were lower than those of female patients at age <21 years, and the cylindrical refraction was higher in males than in females among the patients at age 21~<31 years, and the differences were statistically significant (both at P<0.05). Among the patients with onset age <21 years and diagnosis age <21 years, the ratio of males to females in patients with severe keratoconus was higher than those with mild and moderate disease, and the difference was statistically significant (both at P<0.05). Older age of onset was a protective factor for disease severity in keratoconus (odds ratio=0.981, 95% confidence interval: 0.963~0.999). Conclusions:The younger the onset age of keratoconus patients, the more severe the disease.Among the patients with severe keratoconus, there were more male patients, and males have a younger onset age and severer conditions.It is suggested that early screening of keratoconus in children and adolescents should be strengthened in clinical work, and more active prevention and treatment measures should be taken for younger patients, especially males.

Objective:To investigate the resting state functional connectivity changes of the " triple network model" composed of salient network (SN), executive control network (ECN) and default mode network (DMN) in patients with acute mild traumatic brain injury (mTBI).Methods:From August 2020 to December 2021, forty-five acute mTBI patients (mTBI group) and 40 healthy controls (HC group) with matched sex, age, and education were included.The Montreal cognitive assessment (MoCA) scale was used to evaluate the cognitive status of all subjects.The resting state network (RNS) was established based on independent component analysis (ICA), and the SN, ECN and DMN were extracted, then functional network connectivity (FNC) was analyzed.Subsequently, the correlation between functional connectivity abnormalities and the performance of cognitive impairment was analyzed.SPSS 19.0 was used for statistical analysis and double sample t test was used for comparison between the tow groups. Results:Compared with HC group, mTBI group had enhanced functional connectivity between SN(L-insula) (MNI: x, y, z=-36, 15, 0, t=3.693)and ECN (left superior parietal gyrus, L-SPG) (MNI: x, y, z=-33, -69, 54, t=3.333)(FDR adjust, P<0.05), and decreased functional connectivity between DMN(left superior frontal gyrus, L-SFG) (MNI: x, y, z=-30, 30, 42, t=-4.063)and DMN(L-angular gyrus)(MNI: x, y, z=-21, -66, 33, t=-4.101)(FDR adjust, P<0.05). For FNC analysis, functional network connectivity in SN(IC26)-DMN(IC8) was enhanced in the acute mTBI group and decreased between SN(IC26)-DMN(IC12) and ECN(IC3)-DMN(IC12). The changes of left superior parietal gyrus functional connection were negatively correlated with MoCA score ( r=-0.627, P<0.01), and SN (IC26) -DMN(IC12) connection was positively correlated with MoCA score ( r=0.411, P=0.005). Conclusions:In patients with acute mTBI, the resting functional connectivity changes within and between the networks of the " triple network model" composed of SN, ECN and DMN, and is related to the decline of cognitive function.This will help to better understand the neuropathological mechanism of acute mTBI and post-traumatic cognitive impairment, and may become an effective imaging marker for identifying and predicting cognitive impairment after mTBI.

Objective: To explore the cumulative effects of unintentional injury among children in rural area, in order to provide information for early intervention of unintentional injury. Methods: Through multistage clustering sampling method, 2 109 primary caregivers of students from 8 rural primary and elementary schools of Heilongjiang Province were recruited. Strengths and Difficulties Questionnaire (SDQ), Injury Behavior Checklist (IBC), Perceptions of Risks and Hazards were used to collect as the risk factors, while Perceptions of Risks and Hazards (PSAPQ), Home Observation for Measurement of the Environment (HOME) and Knowledge, Attitude and Practice for Children Unintentional Injury (KAP) were also used as the protective factors. Risk factors index (RFI) and protective factors index (PFI) were computed in the study. Results: The severity of unintentional injury were positively correlated with SDQ, IBC and perceptions of risks and hazards(r=0.15, 0.23, 0.12, P<0.01), and were negatively correlated with HOME, PSAPQ and KAP(r=-0.25, -0.14, -0.09, P<0.01). Hierarchical linear regression showed that the total scores of SDQ, IBC and environmental of HOME predicted the severity of unintentional injury which could explain 34% variant of unintentional injury. It also indicated that the severity of unintentional injury were positively correlated with RFI (β=0.21) and negatively correlated with PFI(β=-0.18), the interaction was significant(β=-0.11,R2=0.31)(P<0.01). Conclusion Both risk and protective factors of unintentional injury have cumulative effects on the severity of injury among rural children. The relationship between risk factors and injury could be mediated by protective factors.

Objective To investigate the expression of Sema4 D and Ki67 in pancreatic carcinoma and its relationship with clinicopathological features. Methods The expression of Sema4 D and Ki67 in 40 pancreatic carcinoma tissues and 10 adjacent tissues was measured by immunohistochemistry, and its relationship with the clinicopathological features of pancreatic carcinoma was analyzed. Comparison of categorical data between two groups was made by chi-square test. Spearman's rank correlation analysis was used to identify the correlation between these variables. Results Sema4 D and Ki67 were detected in 19 (47. 5％ ) and 18 (45. 0％ ), respectively, of the 40 pancreatic carcinoma tissues, with significantly higher positive rates than the adjacent tissues (χ2= 10. 040 and 10. 000, P = 0. 020 and 0. 022) . The abnormal expression of Sema4 D and Ki67 differed significantly between patients with different tumor stages and between patients with and without lymph node metastasis (χ2= 6. 352, 4. 912, 12. 031, and 6. 465, P = 0. 013, 0. 028, 0. 001, and 0. 025) . Sema4 D expression was positively correlated with Ki67 expression (r = 0. 347, P = 0. 028) . Conclusion The expression of Sema4 D and Ki67 is elevated in pancreatic carcinoma compared with the adjacent tissue. Their expression is positively correlated with each other and is associated with tumor staging and lymph node metastasis.

Objective To evaluate the analytical performance of four lipoprotein associated phospholipase A2(Lp-PLA2)activity reagents on Beckman AU5800 automatic biochemical analyzer. Methods The remaining serum samples of 214 patients and 140 apparently healthy individuals were collected from March to July 2017 in Peking Union Medical College Hospital.These samples were used for method comparison and reference interval evaluation.According to the guidelines of EP15-A,EP6-A,EP-17 and EP7-P from Clinical and Laboratory Standards Institute(CLSI)standards,the precision, linearity, sensitivity and common interferences(e.g free bilirubin, conjunct bilirubin, hemoglobin and chyle)were assessed.According to EP9-A2,method comparisons of differents regents(Evermed,DiaSys,Hengxiao and Zhongyuan were labeled as A,B,C and D,respectively)were conducted and the differences were estimated at medical decision levels(328U/L,391U/L and 485U/L).Results The precision of four reagents were acceptable.The repeatability(CV%)of A to D were 0.5%-1.7%, 0.7%-3.0%, 0.9%-2.0% and 0.5%-3.3%,respectively.The reproducibility(CV%)were 0.7%-2.9%, 1.4%-3.2%, 1.3%-1.9%and 0.8%-4.1%,respectively.Both of those achievedlaboratory defined quality objective(<5%).The linearity of A to D were 44 -1 992 U/L,39 -1 798 U/L,13 -540 U/L and 75-1 717U/L,respectively.The regression coefficient R2 was between 0.997 and 1.000, and the correlation coefficient(r)was between 0.998 and 1.000.The interference of chyle were acceptable among these four reagents andmet the manufacturer′s requirementsor clinical needs.In a low level of Lp-PLA2,bilirubin had an obvious interferenceonreagent C;B and C were negatively affected when the hemoglobin was 4.5 g/L; and D was positively affected when the hemoglobin was 2.45 g/L.The regression coefficients R2 of A,C,D compared with B were between 0.978 and 0.995,and the correlation coefficients(r)were between 0.989 and 0.998. The expected differences at medical decision levels ranged from -240 U/L to 113 U/L.For A to D,the Lp-PLA2 activity results of 131(93.6%), 140(100%), 82(58.6%), and 128(91.4%)cases were analysed within the manufacturer′s claimed reference intervals.Conclusion The precision and linearity of the four Lp-PLA2 activity detection reagents used in automatic biochemical analyzer are good, but the anti-interference ability needs to be improved.

Objective: To evaluate the effects of CYP2C19 genetic polymorphism on the plasma concentration of voriconazole in patients with hematological disease and the value of serial monitoring plasma concentrations in the treatment and prevention of invasive fungal disease (IFD). Methods: From January 2016 to December 2016, 65 hematological patients who received voriconazole intravenous administration for the treatment of invasive fungal disease were enrolled in this study. The population CYP2C19 polymorphism of voriconazole were performed using PCR-Pyrosequencing. The trough plasma concentrations of vriconazole (C_trough) was detected by ultra performance liquid chromatography tandem mass spectrometry. Results: Based on the genotype analysis, 65 subjects were identified as extensive metabolizers’ group (30 cases) and poor metabolizers’ group (35 cases). The C_trough of the 65 patients were detected for 169 times totally, and there was a significant difference of C_trough values between the two groups [0.98(0.38-2.08) mg/L vs 2.19(1.53-4.27) mg/L, z=10.286, P<0.001]. The medium of C_trough in 65 hematological patients were described. Lack of response to therapy was more frequent in patients with voriconazole levels <1.5 mg/L (50.0%) than in those with voriconazole levels >1.5 mg/L (20.5%) (P=0.052). And the risk of adverse events was more frequent in patients with voriconazole levels >5.5 mg/L (80.0%) than in those with voriconazole levels ≤5.5 mg/L (8.3%) (χ²=11.689, P=0.020). Conclusion Patients with CYP2C19 wild-type phenotype are extensive metabolizers, their C_trough of voriconazole are significantly lower than patients with CYP2C19 non-wild-type phenotype (poor metabolizers). Appropriate concentrations of vriconazole can improve the efficacy and safety during treatment.

AIM To observe the inhibitory effects of tricin on the airway inflammation in low concentration LPS induced asthma mice.METHODS The model of asthma mice with airway inflammation was induced by low concentration LPS plus OVA.There were four groups:blank control group,asthma model group,tricin treatment group and dexamethasone (positive drug) treatment group.The levels of inflammatory factors,including NO,TNF-α and IL-1β,in bronchoalveolar lavage fluid (BALF) were detected by ELISA;the pathology of lung tissue was observed by HE staining;the mRNA and protein expressions of TLR4,MyD88 and NF-κB p65 in alveolar macrophages were detected using real-time quantitative PCR (RT-PCR) and Western blot.RESULTS Tricin could significantly decrease the levels of NO,TNF-α and IL-1β in BALF of asthma mice and improve lung inflammation.It could also down-regulate the relative mRNA and protein expressions of TLR4,MyD88 and NF-κB p65.CONCLUSION Tricin has inhibitory effects on airway inflammation in asthma mice,whose mechanism may be related to the intervention of TLR4/MyD88/NF-κB pathway.

Objective To evaluate the application value of F wave by comparison with quantitative sensory testing(QST)and nerve conduction studies(NCS)in diabetes peripheral neuropathy (DPN). Methods Three hundred and fourteen diabetic patients with symptom of DPN were retrospectively analyzed, who were recruited and divided into four groups according the course of diabetes mellitus: ≤ 3 years group, 4-6 years group, 7-9 years group and >9 years group. The abnormal rates of QST, NCS and F wave were analyzed and compared. Results At earlier course of disease (≤ 3 years group, 4- 6 years group), the abnormal rate of QST [76.54%(62/81), 91.67% (88/96)]>F wave[(48.15%(39/81),64.58%(62/96)]>NCS[(32.10%(26/81),47.92%(46/96)], and there were significant differences(P<0.01).The abnormal rate of QST, F wave and NCS at 7-9 years group and>9 years group had no significant differences(P>0.05).The abnormal rate of QST in 4-6 years group[(91.67%(88/96)]was higher than that in≤3 years group[(76.54%(62/81)],and decreased in 7-9 years group and >9 years group. The abnormal rate of NCS was increased with course of disease, and the range was obviously between close together group. The abnormal rate of F wave was increased with course of disease,but in 4-6 years group the range was obvious and the range was slow down.At earlier course of disease (≤3 years group, 4-6 years group), the abnormal rate of only F wave significantly increased the abnormal rate of NCS + F wave from 30.86%(25/81), 44.79% (43/96)to 46.91%(38/81), 61.46%(59/96)respectively, but in 7-9 years group and>9 years group, the abnormal rate of only F wave increased the abnormal rate of NCS + F wave from 69.44%(50/72), 84.61%(55/65)to 72.22% (52/72),86.15%(56/65).Conclusions Although the abnormal rate of F wave is not as high as QST, it is still remarkable. Detection of F wave shows positive correlations with the course of disease, which can reflect the course of disease objectively and reliably.The added F wave detection can distinctly raise the abnormal rates on the base of NCS.