[Objective] To investigate the differences in metabolomics between apheresis platelets stored at 4℃ and at 22℃ with agitation, aiming to provide a theoretical basis for the cold storage of apheresis platelets. [Methods] Samples were collected at four time points (d1, d5, d10, d15) for platelets stored at 4℃ (experimental group) and two time points (d1, d5) for platelets stored at 22℃ with agitation (control group). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology was used to detect changes in platelet metabolome levels under different storage conditions. Platelet functional activity was assessed by thromboelastography (TEG) for maximum amplitude (MA) values and flow cytometry for CD62P activation rates. [Results] Metabolites in the glycolytic pathway, key metabolites in the tricarboxylic acid cycle (citrate, α-ketoglutarate), metabolites in the purine metabolism pathway (adenine, inosine monophosphate, guanine, etc.) and amino acid metabolites significantly decreased by d5 in the control group, whereas they remained stable in the experimental group. The content of fatty acid metabolites, such as prostaglandin G2, 13(S)-HOTrE, and linoleic acid, significantly increased in the control group. Statistically significant differences in MA values were observed between the two groups at d1 and d5 (P<0.05). However, in the experimental group, as the storage time extended, the MA values at d10 and d15 showed no significant difference compared to the control group at d5 (P>0.05). The CD62P activation rate between the two groups was statistically significant (P<0.05). Additionally, the CD62P activation rate of platelets in the 22℃ group increased rapidly from d1, while it rose gradually in the 4 ℃ group. [Conclusion] Platelets stored at 4 ℃ exhibit more stable metabolic activity and slower functional deterioration, which is beneficial for extending the effective storage period of platelets.

[Objective] To determine the blood group of a patient with ABO forward and reverse typing discrepancies using PacBio third-generation sequencing (TGS) technology, and to explore the application of serological methods and molecular biological methods in identifying chimeric blood groups. [Methods] The blood group serology testing was utilized. PCR amplification and Sanger sequencing of exons 1-7 of the ABO gene were conducted. The full-length sequencing of the ABO gene and haplotype analysis were carried out by PacBio third-generation sequencing technology. Short tandem repeat typing was also performed. [Results] Serological testing suggested a suspected B subtype, which appeared mixed field of vision with anti-B antibodies and showed 2+mf strength agglutination. Sanger sequencing revealed a homozygous ABO^* O. 01. 01 genotype with the c. 261delG mutation in exon 6. PacBio TGS identified a predominant ABO^* O. 01. 01/ABO^* O. 01. 01 genotype and a low proportion of ABO^* B. 01. Nine locis of twenty short tandem repeat (STR) locis showed three or four types of genotypes in STR analysis, confirming chimerism. [Conclusion] The sample was a B/O blood group chimerism. The low proportion of ABO^* B. 01 chimerism was the true cause for the serological mixed field of vision. The PacBio third-generation sequencing technology can not only determine the ABO gene haplotype but also detect a low proportion gene chimerism in ABO blood groups.

【Objective】To explore the relationship between the miR- 132 expression in serum and cognitive deficits of OSA. 【Methods】 66 Chinese adults age 30 to 60 years old were enrolled and categorized into two groups based on Montreal Cognitive Assessment（MoCA）scores： OSA patients with cognitive impairment（OSAI，n=36），OSA patients without cognitive impairment（OSAN，n=30），and thirty adults without OSA as healthy control group（HC，n=30）. Out- of- center cardiopulmonary sleep testing （OCST） and MoCA assessment were performed and the relative expression of miR-132 in serum was detected by PCR.【Results】No significant difference was observed in age，education，gender and hypertension（P>0.05）. The relative expression level of miR-132 was significantly up-regulated in OSAI patient′s serum compared to the OSAN and HC patients （P<0.001），and had a positive correlation with MoCA score （r=- 0.726，P<0.001）. ROC analyses showed that the areas under the curve（AUC）were statistically significant from the line of identity in OSA with cognitive impairment（AUC=0.935，95% CI：0.890- 0.981，P<0.001）and in OSA（AUC=0.787，95% CI：0.695-0.879，P<0.001）.【Conclusions】Elevated serum miR-132 expression levels are closely related to the diagnosis of OSA and its cognitive dysfunction. Detection of serum miR- 132 may be a potential indicator of cognitive dysfunction and diagnosis in OSA patients.

OBJECTIVE: To explore the effects of acupoint application on pulmonary function and quality of life in patients with pneumoconiosis. METHODS: By random number table method,74 pneumoconiosis patients were randomly divided into two groups: acupoint application group and control group,37 cases in each one. The control group was given conventional treatment. Based onconventional treatment,the acupoint application group was given acupoint application with Chinese herbal medicine on the first day of Chu Fu,Zhong Fu,Mo Fu every year for three years. The changes of pulmonary function was observed in each group. The quality of life of patients was investigated by the World Health Organization Quality of Life-BREF. RESULTS: Before treatment,the pulmonary function indicators such as vital capacity,forced vital capacity(FVC),forced expiratory volume in first second(FEV1),FEV1/ FVC% 、peak expiratory flow( PEF),maximal voluntary ventilation and quality of life score between the two groups had no statistical significance( P > 0. 05). Compared with the same group before treatment,all indexes of pulmonary function,except FEV1/ FVC% and PEF,and life quality score were improved in patients in the control group after treatment( P < 0. 05). After treatment,all indexes of pulmonary function and life quality score were improved in patients in acupoint application group( P < 0. 01). The remaining 5 lung function indexes,except PEF,and the quality of life of the patients in acupoint application group were higher than those in the control group( P < 0. 05). CONCLUSION: Acupoint application therapy can improve lung ventilation function and improve quality of life in patients with pneumoconiosis.

【Objective】 To explore the precautions of pre-transfusion examination in patients with antibodies to erythrocyte protective solution, discrepant ABO blood typing results, and positive unexpected antibodies, so as to ensure the safety of blood transfusion. 【Methods】 The screen cells were divided into two groups according to the presence or absence of washing reagent red blood cells in normal saline. One group had untreated forward typing cells, antibody screening cells and identification panel, and the other group had saline-washed reverse typing cells, antibody screening cells and identification panel. The experiments were carried out by microcolumn gel method, saline medium method and polyamine method to analyze the effect of red blood cell preservation solution on serum agglutination reaction of specific patients. 【Results】 Among the 8 patients, forward typing was AB (+ ) in 1 patient, B (+ ) in 4, and A(+ ) in 3, and the reverse typing were interfered. The plasma of 8 patients agglutinated with unwashed reverse typing cells (saline tube method), screen cells and identification panels (saline tube method plus cassette method), while not agglutinated with the polybrene method. The interference was eliminated as using washed reverse typing cells (salinetube method), screen cells and identification panels (saline tube method plus cassette method). 【Conclusion】 The erythrocyte preservation solution affected patients’ blood group typing, but not affected the outcome of blood transfusion as no adverse reactions occurred.

Objective To evaluate the efficacy and safety of permissive hypercapnia ventilation in mechanically ventilated newborns.Methods PubMed,Embase,the Cochrane Library,China National Knowledge Infrastructure (CKNI),Wanfang Data and Chinese BioMedical Literature Database (CBM) were searched up until March 31,2013.Randomized controlled trials (RCTs) comparing permissive hypercapnia (PHC) group with normocapnia (NC) group in mechanically ventilated newborns were included.The primary outcomes included the incidence of ventilator associated lung injury (VALI),bronchopulmonary dysplasia (BPD),intraventricular hemorrhage (IVH),periventricular leukomalacia (PVL),patent ductus arteriosus (PDA),neonatal necrotizing enterocolitis (NEC),neurodevelopmental injury and the mortality rate.Secondary outcomes included the duration of ventilatory support and the length of hospital stay.The Cochrane Handbook 5.1.0 was used to evaluate the methodological quality and RevMan 5.1 software from Cochrane Collaboration was used for meta-analysis.The fixed effects model or the random effects model was adopted according to the result of heterogeneity.Results (1) A total of 325 articles were searched,and eight RCTs involving 605 newborns (302 newborns in PHC group while 300 newborns in NC group) which met the inclusion criteria were selected.In seven studies,the partial pressure of carbon dioxide (PCO2) was controlled at ＜ 65 mmHg (1 mmHg=0.133 kPa) and pH at ≥ 7.2 in PHC group.In one study,PCO2 was ＞ 52 mmHg and pH＞7.2,without descripition of the upper limit of PCO2.(2) Four articles described the method of random allocation in detail; three described allocation concealment; all eight studies used blinding method for research subjects; two used blinding method for outcome assessment; all eight studies reported complete data; and three articles described the source of other bias.(3) All eight studies reported the incidence of VALI (I2=56％,P=0.03).The random effects model was used for the meta-analysis,and there was significant difference between PHC group and NC group (RR=0.52,95％CI:0.29-0.93,P=0.03).According to the gestational age,the eight studies were divided into two subgroups.One subgroup,including three studies with an average gestational age of 25 weeks (I2=0％,P=0.46),showed no significant difference in the incidence of VALI between PHC and NC group (RR=1.05,95％CI:0.72-1.54,P=0.78).The other subgroup,including five studies with gestational age of ＞27 weeks (I2=0％,P=0.68),showed significant difference in the incidence of VALI between the two groups (RR=0.27,95％CI:0.14-0.50,P＜0.01).The in-hospital mortality and duration of ventilation showed significant difference between the two groups (in-hospital mortality:RR=0.40,95％CI:0.22-0.74,P＜0.01; duration of ventilation:difference in means=-0.75,95％CI:-1.04--0.46,P＜0.01).There was no significant difference in the incidence of BPD,IVH,PVL,PDA,NEC and neurodevelopmental impairment between the two groups (all P＞0.05).Conclusions PHC ventilation in mechanically ventilated newborns can decrease the incidence of VALI,the in-hospital mortality and the duration of ventilation,while its protective efficacy against BPD is not remarkable.It does not increase the risk of IVH,PVL,PDA,NEC and neurodevelopmental injury,when the PCO2 is ＜ 65 mmHg and pH ≥ 7.2.

Objective: To investigate the effects of epimedium water extract (EWE) on the ability of osteogenic differentiation and the expression of TGF-β1 and BMP-2 in inducing mesenchymal stem cells (MSCs) differenting into osteoblast and to further explain its mechanism. Methods: MSCs were isolated and purified by differential time adherent method; the most effective concentration of EWE on the osteogenic differentiation of MSCs was confirmed by the activity of alkaline phosphatase (ALP) and positive rates of ALP staining; according to the differently induced condition, MSCs were divided into four groups: control, classic (induced by the classic osteoblast-induced system), EWE (induced by the most effective concentration of EWE on the osteogenic differentiation), and EWE + classic (induced by the combination of classic osteoblast-induced system and the most effective concentration of EWE on osteogenic differentiation) groups. ALP, type I collagen (Col I), bone gla protein (BGP), and calcium nodes in each group were detected and compared to indicate the osteogenic differentiation of each group. TGF-β1 and BMP-2 in each group were detected by ELISA. Results: The most effective concentration of the EWE on the osteogenic differentiation of MSCs was 500 μg/mL. The classic, EWE, and EWE + classic groups could promote the osteogenic differentiation of MSCs and increase the expression of TGF-β1 and BMP-2. Conclusion: The EWE could promote the osteogenic differentiation of MSCs. The increase of the expression of TGF-β1 and BMP-2 in the induced groups of EWE may be the mechanism of improving the differentiation of MSCs into osteoblast.

OBJECTIVE: To develop an HPLC method for simultaneous determination of multiple-components in Hedyotis diffusa Willd. METHODS: The HPLC analysis was carried out on a C18 column (4.6 mm×250 mm, 5 μm) by gradient elution with acetoni-trile-water[both containing 0.1‰ (V/V) acetic acid] as mobile phase at a flow rate of 0.8 mL·min-1, the column temperature at 35°C, and the detection wavelength was set at 238 nm. External standard method and quantitative analysis of multi-components by single marker (QAMS) method were adopted for simultaneous determination of six components in Hedyotis diffusa Willd, respectively. RESULTS: The linear ranges for asperulosidic acid, quercetin-3-O-[2-O-(6-O-E-feruloyl)-β -D-glucopyranosyl]-β-D-glucopyrano-side, kaempferol-3-O-[2-O-(6-O-E-feruloyl)-β-Z) -gfucopyranosyl]-β-D-galactopyranoside, (E)-6-O-p-coumaroyl scandoside methyl ester, (E)-6-O-feruloyl scandoside methyl ester, (Z)-6-O-p-coumaroyl scandoside methyl ester were 2.34-93.50, 2.61-104.33, 0.67-26.69, 3.42-136.84, 0.65-26.07, and 1.10-44.17 μg·mL-1 (r<0.9993), respectively. The RSD values of precision, reproducibility, and sample stability were not more than 2.2%. The average recoveries of the six components were 99.8%-101.1% with RSDs not more than 1.2%. The P values of external standard method and QAMS by paired t-test were greater than 0.05. CONCLUSION: There is no significant difference in the content analysis results of the two methods, which can both used for simultaneous determination of the four iridoids and two flavonoids in Hedyotis diffusa Willd.

AIM:To observe the changes of macular morphology and microcirculation in myopic maculopathy(MM), and investigate theirs correlation and effects on vision.METHODS: Case-control study. A total of 165 patients(189 eyes)with high myopia and 154 healthy volunteers(154 eyes)from October 2016 to December 2018 were selected. According to the classification of Meta-analysis for pathologic myopia(META-PM), participants were divided into M0 group(category 0, 41 eyes), M1 group(category 1, 53 eyes), M2 group(category 2 and 3, 52 eyes), and myopic choroidal neovascularization(mCNV)group(43 eyes). All participants underwent optical coherence tomography angiography(OCTA)examination. Morphological and microcirculation parameters of retina at different layers were compared between groups. Pearson correlation was used to assess the correlation between morphological and microcirculation parameters. Correlations between vision and other parameters were analyzed using multiple linear regression analysis.RESULTS:Foveal full retinal thickness(FRT)and outer retinal thickness(ORT)were all lower in M0, M1 and M2 groups than those of control group(all P<0.01). Foveal superficial capillary plexus vessel density(SVD)and deep capillary plexus vessel density(DVD)were all lower in M2 and mCNV groups than those of the control group(all P<0.01). Parafoveal FRT and ORT were all lower in M0, M1, M2 and mCNV groups than those of the control group(all P<0.01). Parafoveal inner retinal thickness(IRT), SVD and DVD were all lower in M2 and mCNV groups than those of the control group(all P<0.01). Subfoveal choroidal thickness(SFCT)and choroid capillaries vessel density(CVD)were all lower in M0, M1, M2 and mCNV groups than those of the control group(all P<0.01). Foveal vessel density of retina and choroid were positively correlated with its thickness in patients with MM without CNV(all P<0.05). Multivariate analysis showed that axial length(AL), diffuse or patchy chorioretinal atrophy were influencing foctors of best corrected visual acuity(BCVA; all P<0.01).CONCLUSION:Retinal morphological changes precede microcirculation changes in MM. Most of all, ORT changes precede IRT changes. Foveal vessel density of retina and choroid were positively correlated with its thickness. The main influencing factors of BCVA were AL and types of MM.

Objective: To investigate the effects of transcription factor 21 (TCF21) on the proliferation, apoptosis, invasion and migration and angiogenesis of human breast cancer cells (MDA-MB-231). Methods: TCF21 overexpressing plasmid was transfected into human breast cancer cell line MDA-MB-231 by liposome transfection, and Western blot was used to detect whether the transfection was successful. The effects of TCF21 overexpression on proliferation, apoptosis, invasion and migration and angiogenesis of MDA-MB-231 were detected by MTT, DAPI, Transwell and CAM. Results: The expression of TCF21 protein in Western blot showed that the TCF21 overexpression plasmid was successfully transfected into MDA-MB-231 cells. Overexpression of TCF21 inhibited the proliferation of tumor cells and inhibit the proliferation of 48 h cells after transfection. Overexpression of TCF21 inhibited tumor invasion and migration and angiogenesis, and promoted apoptosis of tumor cells. The results of Western blot showed that the protein expression of MMP-9, VEGFA and p-VEGFR2 decreased significantly after overexpression of TCF21. Conclusions TCF21 can inhibit the proliferation, invasion and migration, angiogenesis and apoptosis of MDA-MB-231 cells. It is suggested that TCF21 can be used as a potential site for clinical diagnosis and treatment of breast cancer.