[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 study the reasons for ambiguous blood matching in a patient who received multiple blood transfusions,and explore how to reduce the risk of producing homologous antibodies in patients who received multiple blood transfusions.Methods The antibody specificity was detected by serum antibody screening and identification,polyethylene glycol(PEG)immunoglobulin assay and dimercaptoethanol method based on clinical data analysis.Results The unexpect-ed antibodies in the patient's serum were anti-C and anti-Wra,and anti-Jkb was detected after using polyethylene glycol im-munoglobulin enhancement experiments.The anti-Wra was detected as IgM by dimercaptoethanol method,with a titer of 16,anti-C of 8,and anti-Jkb＜1.Conclusion One case of ambiguous blood matching caused by anti-C,anti-Jkb combined with anti-Wra was discovered.By strengthening the experiments,the detection efficiency of Kidd system antibodies was improved,enabling timely provision of matching blood to patients.

Objective:To explore the effect of intervention model based on comprehensive nutrition management on glucose and lipid metabolism and pregnancy outcome in patients with gestational diabetes mellitus (GDM).Methods:104 GDM patients admitted to Shanxi Bethune Hospital from February 2022 to March 2023 were randomly divided into control group and experimental group, with 52 cases in each group. The control group implemented routine management measures and nutrition guidance, while the experimental group implemented an intervention model based on comprehensive nutrition management on the basis of routine management. The indexes of glucose and lipid metabolism (glycosylated hemoglobin, fasting blood glucose, 2 h postprandial blood glucose, total cholesterol, triglyceride and low density lipoprotein cholesterol), pregnancy outcome, self-management ability and self-efficacy were compared between the two groups before and after intervention.Results:Before the intervention, there was no significant difference in general situation, glucose and lipid metabolism index, self-management ability and self-efficacy between the two groups ( P>0.05). After the intervention, the level of glucose and lipid metabolism index in the experimental group was significantly lower than that in the control group, with statistical significance ( P<0.05). The incidence of adverse pregnancy outcome in the experimental group was significantly lower than that in the control group, with statistical significance ( P<0.05). The scores of self-management ability and self-efficacy in the experimental group were significantly higher than those in the control group, with statistical significance ( P<0.05). Conclusion:The intervention model based on comprehensive nutrition management can effectively improve the glucose and lipid metabolism index and pregnancy outcome of GDM patients, and significantly improve their self-management ability and self-efficacy related to nutrition management, which has high clinical application and promotion value.

Objective:To observe the safety and efficacy of 0.01% atropine eye drops in the prevention of myopia onset in schoolchildren.Methods:A randomized double-blind controlled study was conducted.Sixty Chinese Han children (60 eyes) with binocular spherical equivalent (SE) between + 0.50 D and -0.75 D (pre-myopia) by cycloplegic autorefraction treated in The First Affiliated Hospital of Zhengzhou University were enrolled from July to October 2020.Aged 6-12 years old, the children were divided into 0.01% atropine group and control group according to a random number table, with 30 cases (30 eyes) in each group.The children were given one drop of 0.01% atropine or placebo eye drops in both eyes once a night.The SE, axial length (AL), accommodative amplitude and pupil diameter were compared before and after 3-month, 6-month of treatment between the two groups.Discomforts were recorded.This study adhered to the Declaration of Helsinki.The study protocol was approved by an Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2020-KY-286). Written informed consent was obtained from guardian of each subject.Results:After treatment, 26 and 25 subjects completed the 6-month follow-up in 0.01% atropine group and control group, respectively, among which 3 subjects in 0.01% atropine group accounting for 11.5% and 9 in control group accounting for 36.0% developed myopia, showing a statistically significant difference ( χ2=4.238, P=0.040). There were significant differences in the overall comparison of SE and AL at different time points between before and after treatment ( Ftime=10.981, 81.854; both at P<0.001). At 3 and 6 months after treatment, there were significant increases in the SE and AL of control group and AL of 0.01% atropine group compared with respective baseline values (all at P<0.05). There was no significant difference in SE at 3 and 6 months after treatment compared with baseline SE in 0.01% atropine group (both at P>0.05). At 6 months after treatment, the change in SE in 0.01% atropine group was (-0.15±0.26)D, which was significantly less than (-0.34±0.35)D in control group, and the change in AL in 0.01% atropine group was (0.17±0.11)mm, Which was significantly shorter than (0.28±0.14)mm in control group, with significant differences between them ( t=2.207, P=0.032; t=3.127, P=0.003). There were significant differences in pupil diameter at different time points between before and after treatment ( Ftime=2.263, P=0.032). At 3 and 6 months after treatment, the pupil diameter was increased in comparison with baseline in 0.01% atropine group (both at P<0.05). There were significant differences in accommodative amplitude at different time points between before and after treatment in the two groups ( Fgroup=0.882, P=0.042; Ftime=0.337, P=0.033). The accommodative amplitude at 3 and 6 months after treatment were decreased in comparison with baseline in 0.01% atropine group and control group at corresponding time points (all at P<0.05). Within a month after treatment, photophobia in bright sunlight occurred in 5 cases in 0.01% atropine group, accounting for 16.7%(5/30), and 2 cases in control group, accounting for 6.7%(2/30), showing no significant difference ( χ2=0.647, P=0.421). No near-vision blur and other uncomfortable symptoms was found in the two groups. Conclusions:After 6-month application of 0.01% atropine eye drops, the prevalence of myopia in pre-myopia schoolchildren decreases and the changing rate of SE and AL slows down.The accommodative amplitude is slightly reduced and pupil diameter is slightly increased, with no obvious effects on study and life.

Objective:To investigate the effect of long non-coding RNA 068 (lncRNA 068) on the migration of a melanoma cell line A375, and to explore its mechanism of action.Methods:From December 2015 to November 2020, 21 patients with pathologically confirmed cutaneous melanoma were collected from Department of Dermatology, Affiliated Hospital of Nantong University, and quantitative PCR (qPCR) was performed to determine the expression of lncRNA 068 in melanoma and paracancerous tissues. LncRNA 068 was overexpressed or knocked down via lentiviral transfection in A375 human melanoma cells in the following experiments. Specifically, A375 cells were divided into lentiviral vector (LV) -green fluorescent protein (GFP) group and LV-lncRNA 068 group to be transfected with a GFP-expressing LV and a LV containing lncRNA 068 respectively in the overexpression experiment, and were divided into LV-LacZ short hairpin RNA (shRNA) group and LV-lncRNA 068 shRNA group to be transfected with a LV containing the reporter gene LacZ-specific shRNA and a LV containing the lncRNA 068-targeting shRNA respectively in the low-expression experiment, with the LV-GFP group and LV-LacZ shRNA group serving as the control groups. Transwell and scratch assays were performed to evaluate cell migration, EdU cell proliferation assay and cell counting kit-8 (CCK8) assay to determine the proportion of proliferative cells and cell viability respectively, and immunofluorescence staining was conducted to evaluate epithelial-mesenchymal transformation in the above groups. Lentivirus-transfected A375 cells from the above groups were inoculated into the axillae of BALB/c nude mice, and tumor volume was measured and calculated every 3 days. After 30 days, all mice were sacrificed, and tumor tissues were resected to measure the tumor volume and weight. Cultured B16F10 cells were subcutaneously inoculated into the back and foot of BALB/c nude mice to construct mouse models of subcutaneously transplanted B16F10 melanoma. After 2 weeks, the mice were sacrificed, and qPCR and Western blot analysis were performed to determine the mRNA expression of inflammatory factors in transplanted B16F10 melanoma and paracancerous tissues, and expression of IκB kinase (IKK) /P65 signaling pathway-related proteins, respectively. Comparisons between 2 groups were done by t test, and comparisons of tumor volume and weight at different time points among groups were done by repeated measures analysis of variance. Results:qPCR showed that the relative expression of lncRNA 068 was significantly lower in human melanoma tissues and transplanted B16F10 murine melanoma tissues (0.414 ± 0.109, 0.717 ± 0.041, respectively) than in the corresponding paracancerous tissues (1.050 ± 0.103, 1.011 ± 0.023, t = 19.48, 10.83, respectively, both P < 0.001). Transwell and scratch assays both showed that the cellular migratory ability was significantly lower in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.01), and significantly higher in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Immunofluorescence assay showed significantly increased fluorescence intensity of E-cadherin and decreased fluorescence intensity of N-cadherin in the LV-lncRNA 068 group compared with the LV-GFP group (both P < 0.001), but significantly decreased fluorescence intensity of E-cadherin and increased fluorescence intensity of N-cadherin in the LV-lncRNA 068 shRNA group compared with the LV-LacZ shRNA group (both P < 0.05). In vivo tumor formation experiment in nude mice showed that there were no significant differences in the volume or weight of melanoma between the LV-lncRNA 068 group and LV-GFP group (both P > 0.05), as well as between the LV-lncRNA 068 shRNA group and LV-LacZ shRNA group (both P > 0.05). As qPCR and Western blot analysis showed, the mRNA and protein expression of interleukin-10 (IL-10) and claudin-1 in A375 cells were significantly higher in the LV-lncRNA 068 group than in the LV-GFP group (both P < 0.05), but significantly lower in the LV-lncRNA 068 shRNA group than in the LV-LacZ shRNA group (both P < 0.05). Compared with the paracancerous tissues, B16F10 melanoma tissues showed significantly decreased mRNA expression of IL-10 ( P < 0.01), but significantly increased mRNA expression of IL-6 and tumor necrosis factor-α, as well as protein expression of phosphorylated P65 and phosphorylated IKK ( P < 0.01) . Conclusion:Overexpression of lncRNA 068 can inhibit the migration of A375 melanoma cells, and may affect the development of inflammation and inhibit the epithelial-mesenchymal transformation by inhibiting the IKK/P65 signaling pathway.

Knee osteoarthritis (KOA) is a kind of degenerative osteoarthrosis, which usually involves articular cartilage, subchondral bone, meniscus and quadriceps femoris, which is equivalent to the change of "tendon" in traditional Chinese medicine. The occurrence and development of the change of "tendon" can be explained by the theory of "bone-restoration and tendon-softening" in traditional Chinese medicine. If the force line of the lower limb of the knee joint is normal, it can be indicated by "bone-restoration" in traditional Chinese medicine, that is, the blood around the knee joint runs smoothly, and the "tenons" such as meniscus, ligaments, cartilage, etc. are softened by the blood. This condition of the knee joint can be manifested by the balance of the surrounding soft tissue state. If the force line of the lower limb of the knee joint changes, showing a medial offset and misalignment of the knee joint, it can be indicated by "bone is out of alignment" in traditional Chinese medicine, that is, the structure of the soft tissue around the knee joint and its mechanical characteristics have changed. This condition of the knee joint can be manifested by "the tendons lose their flexibility". In this paper, the theories of modern biomechanics and "bone-restoration and tendon-softening" in traditional Chinese medicine were comprehensively analyzed, the characteristics of the "tenons" when knee osteoarthritis occurs were analyzed, and the characteristics of the occurrence and development of knee osteoarthritis from the perspective of Chinese and Western medicine were discussed.

Porcelain tooth technology is widely used in the treatment of oral diseases, but there are few reports on the possible occupational hazard factors in the process of porcelain tooth production. Porcelain teeth production will produced a large amount of silica dust and metal dust during the grinding process. The technical workers who have been engaged in this work for a long time are very prone to pneumoconiosis due to their poor personal protection awareness. This paper analyzed the clinical data of a pneumoconiosis patient engaged in porcelain tooth making, and analyzed the possible occupational hazard factors in the process of porcelain teeth production, so as to improve the understanding of relevant enterprises, technical workers and medical personnel on the disease and reduce the risk of porcelain teeth production workers suffering from pneumoconiosis.

Porcelain tooth technology is widely used in the treatment of oral diseases, but there are few reports on the possible occupational hazard factors in the process of porcelain tooth production. Porcelain teeth production will produced a large amount of silica dust and metal dust during the grinding process. The technical workers who have been engaged in this work for a long time are very prone to pneumoconiosis due to their poor personal protection awareness. This paper analyzed the clinical data of a pneumoconiosis patient engaged in porcelain tooth making, and analyzed the possible occupational hazard factors in the process of porcelain teeth production, so as to improve the understanding of relevant enterprises, technical workers and medical personnel on the disease and reduce the risk of porcelain teeth production workers suffering from pneumoconiosis.

Objective:To investigate the changes of expressions of NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in the peripheral blood mononuclear cells and downstream factors interleukin-1β (IL-1β) and interleukin-18 (IL-18) in serum in the children with asthma, and to explore their significances on assessing the condition of the children.Methods:A total of 176cases of children with asthma were divided into acute exacerbation group (n=91) , chronic persistent group (n=49) and clinical remission group (n=36) according to the clinical manifestation.During the same period, 60healthy children were selected from the outpatient physical examination center as control group.The pulmonary function of children was checked with lung function instrument.The expression levels of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC) and cysteinyl aspartate-specific proteinase-1 (Caspase-1) mRNA in peripheral blood mononuclear cells of the subjects in various groups were detected by using real-time quantitative PCR.The serum levels of IL-1βand IL-18of the subjects in various groups were detected by using enzyme-linked immunosorbent assay (ELISA) .Results:Compared with control group, the forced expiratory volume in 1second percentage of predicted value (FEV1％) and fixed ratio of forced expiratory volume in the first second/forced vital capacity (FEV1/FVC) of the children in acute exacerbation, chronic persistent and clinical remission groups were decreased (P＜0.05) ;acute exacerbation group＜chronic persistent group＜clinical remission group, and there were significant differences between various groups (P＜0.05) .The levels of NLRP3, ASC and Caspase-1mRNA in the peripheral blood mononuclear cells and serum levels of IL-1βand IL-18in the children with asthma were higher than those in control group (P＜0.01) .The expression levels of NLRP3, ASC and Caspase-1mRNA in the peripheral blood mononuclear cells of the children in acute exacerbation group were higher than those in chronic persistent and clinical remission groups (P＜0.05) , and the expression levels of NLRP3, ASC and Caspase-1mRNA in chronic persistent group were higher than those in clinical remission group (P＜0.05) .Pearson correlation analysis showed that the expression level of NLRP3mRNA in the peripheral blood mononuclear cells of asthmatic children was positively correlated with the expression levels of ASC, Caspase-1 mRNA and the serum levels of IL-1βand IL-18 (P＜0.05) , while it was negatively correlated with FEV1％and FEV1/FVC;the expression level of ASC mRNA was positively correlated with the expression level of Caspase-1mRNA and the serum levels of IL-1βand IL-18 (P＜0.05) , while it was negatively correlated with FEV1％and FEV1/FVC (P＜0.05) ;the expression level of Caspase-1 mRNA was positively correlated with the expression level of Caspase-1mRNA and the serum levels of IL-1βand IL-18 (P＜0.05) , while it was negatively correlated with FEV1％and FEV1/FVC (P＜0.05) ;the serum level of IL-1βwas negatively correlated with FEV1％and FEV1/FVC (P＜0.05) , and the serum level of IL-18was negatively correlated with FEV1％and FEV1/FVC (P＜0.05) .Conclusion:The expression levels of NLRP3inflammasome and the downstream factor IL-1βand IL-18in peripheral blood of the children with asthma are increased, and they are related to the clinical stage of the children with asthma.NLRP3inflammasome pathway might promote the pathogenesis of asthma in the children.

Objective To analyze the clinical value of digital breast tomosynthesis (DBT) compared with digital mammography (DM) and ultrasound for diagnosing non-calcified masses in dense breasts.Methods Images taken with DBT,DM and ultrasound of 1144 patients with non-calcified masses in dense breasts were retrospectively analyzed using breast imaging reporting and data system (BI-RADS).Taking histopathologic results as golden standards,the detection rate and diagnostic accuracy,sensitivity,specificity,false negative and BI-RADS category were evaluated and compared statistically.Results The detection rate of DBT,DM and ultrasound for non-calcified massed in dense breasts was 86.62％ (991/1 144),77.80％ (890/1 144) and 99.65％ (1 140/1 144),respectively (P＜0.05),while the diagnostic accuracy was 83.92％ (960/1 144),75.00％ (858/1 144) and 94.67％ (1 083/1 144),respectively (P＜0.01).The sensitivity of DBT,DM and ultrasound was 89.39％ (312/349),79.93％ (231/289) and 92.70％ (432/466),the specificity was 81.51％ (648/795),73.33％ (627/855) and 96.02％ (651/678),while the false negative rate was 10.60％(37/349),20.07％ (58/289) and 7.30％ (34/466),respectively.No significant difference was found for benign lesions among three examination methods (P=0.75),while there was significant difference for malignant lesions among three examination methods (P＜0.01),and the differences of ultrasonography with DM and DBT,DBT and DM in the for BI-RADS category of malignant lesions were statistically significant (all P＜0.016 7).Conclusion For suspected masses in dense breasts,DBT shows significant advantage than DM,while DBT has the similar advantage compared with ultrasound for the detection and diagnosis of non-calcified masses in dense breasts.