[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 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.

BACKGROUND:Studies on the inhibitory mechanism of Avastin for proliferative diabetic retinopathy are mostly confined to the vascular endothelial growth factor, while connective tissue growth factor and angiogenesis inhibitor also play an important role in the disease.
OBJECTIVE:To evaluate the changes of cytokines in the aqueous undergoing Avastin intravitreous injection or not prior to pars plana vitrectormy in the treatment of proliferative diabetic retinopathy.
METHODS:Thirty patients with proliferative diabetic retinopathy (30 eyes) were selected and randomly divided into groups A, B and C, each group containing 10 patients. Group A was treated with single pars plana vitrectormy;and group B and group C had 0.05 mL/1.25 mg and 0.1 mL/1.25 mg Avastin via intravitreous injection at 7 days before pars plana vitrectormy, respectively. Aqueous humor samples were col ected during pars plana vitrectormy. The levels of vascular endothelial growth factor, connective tissue growth factor and pigment epithelium-derived factor were assessed by using ELISA technique.
RESULTS AND CONCLUSION:The levels of connective tissue growth factor and pigment epithelium-derived factor in the aqueous humor of two Avastin injection groups were increased significantly compared to the group with single pars plana vitrectormy. However, the level of vascular endothelial growth factor was lower in the Avastin injection groups. In addition, there was no significant difference between two Avastin injection groups. These results reveal that intravitreous injection of Avastin assisted vitrectomy for proliferative diabetic retinopathy is more effective than single pars plana vitrectormy treatment. The underlying mechanisms may be that Avastin could inhibit the intraocular angiogenesis by not only reducing the secretion of some growth factors but also increasing the levels of anti-angiogenesis related cytokines.

BACKGROUND:Studies on the inhibitory mechanism of Avastin for early diabetic retinopathy rats are mostly confined to the vascular endothelial growth factor, while connective tissue growth factor and pigment epithelium-derived factor also play an important role in the disease.
OBJECTIVE:To evaluate the changed cytokines of aqueous humor and significance of Avastin intravitreous injection in early diabetic retinopathy rats.
METHODS:Rat models of early diabetic retinopathy were established by inducing with streptozotocin for 10 weeks. Avastin (1.25 mg and 2.5 mg) and physiological saline were injected in the vitreous space.
RESULTS AND CONCLUSION:Enzyme linked immunosorbent assay results revealed that compared with the physiological saline injection group, mass concentration of vascular endothelial growth factor was reduced, mass concentrations of pigment epithelium-derived factor and connective tissue growth factor was increased in aqueous humor of the two Avastin injection groups (P<0.05), but no significant difference in concentration of above cytokines was detected between the two groups (P>0.05). Results verified that Avastin intravitreous injection decreased vascular endothelial growth factor levels, decreased pigment epithelium-derived factor mass concentration and increased connective tissue growth factor levels in rat models of early diabetic retinopathy.

【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.

OBJECTIVES: To analyse the genetic polymorphism of 21 autosome STR loci in Han population of Shandong Province and the cases with loci mutation or allelic loss typed by Goldeneye® DNA identification system 25A. METHODS: Totally 40 autosome STR loci types of 273 unrelated individuals in Han population of Shandong Province were typed by Goldeneye® DNA identification system 25A and 22NC, and the genetic polymorphism of 21 STR loci in those was analysed. Meanwhile, six cases with loci mutation were analysed by adding the tests with Goldeneye® DNA identification system 22NC, 20Y and 17X. Another three cases with allelic loss were tested by AmpFℓSTR® Identifiler® Plus PCR and analysed by gene sequencing. RESULTS: The genetic parameters of 21 autosome STR loci in Han population of Shandong Province were obtained. When STR loci were added up to 40, five of those with loci mutation met the identification requirements, and the results of X-STR or Y-STR types were consistent with that of STR loci. There was another duo case with one suspected loci mutation, biological source of six STR loci genotypes could not be found in the genotypes of supposed father. The Y-STR genotype of two individuals was identical that indicated both of them came from same paternal line. However, the fatherhood was excluded according to the autosome STR loci system. For two cases with allelic loss on D18S51, base mutation or loss were found in the primer binding domain of mother and child by gene sequencing. Another mother-child case with allelic loss on D13S317 was certified by AmpFℓSTR® Identifiler® Plus PCR kit. CONCLUSIONS The 21 autosome STR loci in Han population of Shandong Province have high polymorphism, which can be used in routine cases of paternity identification. For some duo cases with loci mutation, Goldeneye® DNA identification system 25A cannot satisfy the identification requirements, thus more autosome STR loci should be added properly. For the cases with allelic loss, the problem can be resolved by gene sequencing or using different merchant kits.
Asian People/genetics* ; China ; Gene Frequency ; Genetics, Population ; Genotype ; Humans ; Loss of Heterozygosity ; Microsatellite Repeats ; Mutation/genetics* ; Paternity ; Polymerase Chain Reaction ; Polymorphism, Genetic

AIM: To evaluate the safety and efficacy of ultrasonic cycloplasty(UCP)combined with anti-vascular endothelial growth factor(VEGF)+ panretinal photocoagulation(PRP)in the treatment of advanced neovascular glaucoma(NVG).METHODS: Retrospective study. A total of 45 patients(45 eyes)with advanced NVG who received surgery in our hospital from August 2020 to September 2022 were collected and divided into UCP+ anti-VEGF +PRP group(16 patients, 16 eyes), transscleral cyclophotocoagulation(TCP)+anti-VEGF+PRP group(20 patients, 20 eyes), UCP alone group(9 patients, 9 eyes). The intraocular pressure, pain scores, postoperative medication, effective rate, total success rate and the incidence of complications of the patients in the three groups were compared before surgery and at 1 d, 1 wk, 1 and 3 mo after surgery.RESULTS: There was no significant difference in preoperative intraocular pressure, pain scores and preoperative medication of patients in the three groups(all P>0.05). While there were statistical significance in the intraocular pressure and pain scores at 1 d, 1 wk, 1 and 3 mo after surgery(all P<0.01). The intraocular pressure of the UCP alone group(31.78±10.23 mmHg)was found to be higher than that of both the UCP+ anti-VEGF +PRP group(19.44±8.23 mmHg)and the TCP+ anti-VEGF +PRP group(20.80±10.27 mmHg)at 1 mo postoperatively(all P<0.017). The pain score of the TCP+ anti-VEGF +PRP group at 1 d and 1 wk postoperatively was higher than both the UCP+ anti-VEGF +PRP group and the UCP alone group(all P<0.017). The effective rates of UCP+ anti-VEGF +PRP group, TCP+ anti-VEGF +PRP group and UCP alone group were 81%(13/16), 75%(15/20)and 67%(6/9), respectively,(P=0.675), and the success rates were 69%(11/16), 50%(10/20), and 0(0/9), respectively(P=0.003). There was no significant difference in complications of patients in the three groups(P>0.05).CONCLUSION: UCP combined with anti-VEGF +PRP and TCP combined with anti-VEGF +PRP showed comparable efficacy in reducing intraocular pressure in advanced NVG. UCP combined with anti-VEGF+PRP was more effective in relieving pain and with no serious complications in advanced NVG. UCP alone can effectively control intraocular pressure and alleviate the pain of patients in the early postoperative period, but long-term control still requires anti-VEGF+PRP.

Objective To analyse the X-ray and CT manifestations of sacral primary tumor.Methods 10 cases with sacral primary tumors verified by pathology,including sacral chordoma(n=5),giant cell tumor(n=2),chondrosarcoma(n=2),neurilemmomas(n=1),were analysed retrospectively.Results (1)The manifestations of CT and X-ray of sacral chordoma presented as a midline destructive lesion,involving prominently at sacrum 3～5,with flocculent calcification and irregular bone crests.(2)Giant cell tumor involved predominately sacrum 1～3 with expansile destructive area and commonly in young adults.(3)Chondrosarcoma and neurilemmomas had difference characteristic on X-ray film and CT.One case with chondrosarcoma had bone destruction in sacrum 1～3 and giant mass grow into pelvis.Conclusion CT combined with X-ray film is helpful in diagnosis and differential diagnosis of sacral primary tumors.

Objective To explore the protective effect of trimetazidine on the myocardial tissue of rats by observing the oxidative stress, apoptosis and energy metabolism in myocardial tissue of rats after exhausted exercise. Methods Thirty healthy adult male Wistar rats were randomly assigned to three groups (10 each): quiet control group (C), exhausted exercise group (E) and exhausted exercise plus trimetazidine group (TE). Exhausted exercise model was established by forcing the rats swim ceaselessly, rats in group TE received hydrochloric acid trimetazidine (10mg/kg) for intervention. The myocardial tissue of rats was collected after the last exhausted exercise. The myocardial tissue of rats was evaluated by HE staining. The concentrations of superoxide dismutase (SOD), glutathione peroxidase 1 (GSH-Px) and malondialdehyde (MDA) in rats' myocardial cell mitochondria were detected by spectrophotometry. RT-PCR and immunohistochemistry were performed to quantify the mRNA and protein levels of Bax, Bcl-2 and uncoupling protein 2 (UCP2) in myocardial tissue of rats. Terminal-deoxynucleoitidyl Transferase Mediated Nick End Labeling (TUNEL) assay was employed to detect the apoptotic rate of myocardial tissue. Results Compared with group C, the SOD and GSH-Px activities in groups E and TE were reduced (P<0.01), while the MDA activity was increased (P<0.01), and more obvious changes were in group E. The SOD and GSH-Px activities decreased, while the MDA activity increased in group E than in group TE (P<0.01). Compared with group C, the apoptotic rate of myocardial tissue increased in groups E and TE (P<0.01), and the highest apoptotic rate was in group E (P<0.01). Furthermore, the mRNA and protein levels of Bax were significantly elevated in groups E and TE (P<0.01), and the highest expression was in group E (P<0.01). The mRNA and protein levels of BCL-2 were significantly reduced in groups E and TE (P<0.01), and the lowest expression was in group E (P<0.01). Meanwhile, compared with group C, the mRNA and protein expression of UCP2 was significantly increased in groups E and TE (P<0.01). After treatment with hydrochloric acid trimetazidine, the UCP2 expression in group TE was relative decreased than in group E (P<0.01). Conclusion Trimetazidine can reduce the oxidative stress and apoptotic rate of myocardial tissue of rats after exhausted exercise, indicating it plays an important protective role for the myocardial tissue of rats.