[Study on the inhibitory and pro-apoptotic effects of different concentrations of total tanshinone alone and in combination with tyrosine kinase inhibitors on human myeloid leukemia cell lines].

Objective: To explore the effect and investigate the molecular mechanism of different concentrations of total tanshinones alone and in combination with tyrosine kinase inhibitors (TKIs) on the proliferation inhibition and apoptosis of human myeloid leukemia cell lines. Methods: K562 and Kasumi-1 cell lines were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, and the TKIs-resistant strain K562/T315I cell line was constructed in Molecular Medicine Research Center, Beijing Lu Daopei Institute of Hematology. Logarithmic growth phase cells were taken and divided into intervention groups with total tanshinone of 0, 2.19, 4.38, 8.75, 17.50 and 35.00 µg/ml intervention groups, which were inoculated in 96-well plates at a density of 1×104 cells/well and exposed to the drug for 24 h, and a control group treated with dimethyl sulfoxide was also set up simultaneously. All experiments were repeated independently 3-5 times. The proliferative activity of the cells was assessed using the CCK-8 assay, the apoptotic rates were measured by flow cytometry, and the expression levels of apoptosis-regulating proteins Bcl-2 and Bax were analyzed by Western blotting. The cell lines treated and untreated with total tanshinone were subjected to transcriptome sequencing and gene set enrichment analysis to identify differentially expressed genes. Results: The half-inhibitory concentration (IC50) values of 8.75 µg/ml total tanshinone at 24 h for K562, K562/T315I and Kasumi-1 cells were (4.11±0.02), (4.95±0.04) and (3.98±0.01) µg/ml, respectively. When combined with 0.25 µmol/L imatinib, 8.75 µg/ml total tanshinone could enhance the induction of apoptosis effects on K562 and K562/T315I cell lines. After being treated with 4.38, 8.75, and 17.50 µg/ml of total tanshinone for 24 h, compared with the control group, total tanshinone upregulated the expression level of Bax protein, downregulated the expression level of Bcl-2 protein, and decreased the Bcl-2/Bax ratio (all P<0.05). Total tanshinone inhibited the proliferation-related signaling pathway and DNA damage repair pathway of myeloid leukemia cell lines, and activated the signaling pathway that induces apoptosis in leukemia cells. Conclusion: Different concentrations of total tanshinoneinhibites proliferation and promote apoptosis in K562, Kasumi-1 and TKIs-resistant K562/T315I cell lines, and further enhance the anti-leukemic effect when combined with TKIs.

[Safety of delayed vaccination with the national immunization program vaccines in children aged 0-6 years from 2019 to 2021 in Xuhui District, Shanghai City in China].

Objective: To understand the incidence of delayed vaccination with the national immunization program vaccines among children aged 0-6 years in Xuhui District, Shanghai, and to evaluate the safety of delayed vaccination. Methods: A stratified random sampling was used to obtain six vaccination clinics in Xuhui District, Shanghai. The vaccination records of children 0-6 years from these six vaccination clinics were collected from the Shanghai Immunization Program Information Management System. Adverse events following immunization (AEFI) data were collected from the China Information System for Disease Control and Prevention. Descriptive epidemiology was used to analyze the data. Children were divided into the timely vaccination group and delayed vaccination group according whether they were delayed in vaccination (received one month or more after the recommended age among children aged ≤1 year; received three months or more after the recommended age among children aged >1 year). The safety of four vaccination methods-individual vaccination, simultaneous vaccination, routine vaccination and combined vaccination-were further compared. Differences between groups were compared using chi-square test. Results: From 2019 to 2021, six vaccination clinics in Xuhui District administered 124 031 doses of the national immunization program vaccines among children aged 0-6 years, and delayed vaccinations accounted for 25.99% (32 234/124 031) of these doses. In 2020, the delayed vaccination rate during the first-level COVID-19 public health emergency response period in Shanghai was significantly higher than that in the same period in 2019 (34.70% vs. 24.19%, χ2=136.23, P<0.05). The delayed vaccination rate during the COVID-19 vaccination campaign in 2021 was significantly higher than that in the same period in 2019 (25.27% vs. 22.55%, χ2=82.80, P<0.05). From 2019 to 2021, a total of 475 cases of AEFI were reported in six vaccination clinics, with a reported incidence of 382.97 per 100 000 doses, including 421 cases of common adverse reaction (88.63%, 339.43 per 100 000 doses), 51 cases of rare adverse reaction (10.74%, 41.12 per 100 000 doses) and 3 cases of coincidences (0.63%, 2.42 per 100 000 doses). The reported incidence of AEFI among delayed vaccinations was significantly lower than that among timely vaccinations (291.62 per 100 000 doses vs. 415.05 per 100 000 doses). The incidence of AEFI for the four delayed vaccination methods (individual vaccination, simultaneous vaccination, routine vaccination and combined vaccination) was lower than that for timely vaccination. There were significant differences between the groups except for the routine vaccination group (χ2=9.82, P<0.05; χ2=5.46, P<0.05; χ2=2.97, P>0.05; χ2=11.89, P<0.05). Conclusions: In Xuhui District of Shanghai, 25.99% of doses of the national immunization program vaccines administered to children 0-6 years were delayed. Delayed vaccination does not increase the risk of AEFI compared with timely vaccination.

[Early results of left ventricular assist device implantation for the treatment of heart failure].

Twenty-four male patients who underwent left ventricular assist device (LVAD) implantation due to advanced heart failure in Union Hospital, Fujian Medical University from June 2019 to June 2022 were retrospectively included. The age of patients was 32-61 (48.4±8.4) years. Everheat-Ⅰ, HeartCon and Corheart 6 left ventricular assist systems were used in 10, 6 and 8 cases, respectively. All patients were discharged successfully without mechanical failure, thrombosis or secondary thoracotomy for hemostasis. Early postoperative hemodynamics were significantly improved, left ventricular systolic diameter was reduced, left ventricular ejection fraction was gradually improved, and no hemolysis occurred. The patients were followed up for 3 to 39 (17.9±8.6) months, the cardiac function was restored to grade Ⅰ to Ⅱ, and the 6-minute walking test distance increased significantly. Therefore, satisfactory early results can be achieved with left ventricular assist device implantation for the treatment of heart failure.

Observation of a singular Weyl point surrounded by charged nodal walls in PtGa.

Constrained by the Nielsen-Ninomiya no-go theorem, in all so-far experimentally determined Weyl semimetals (WSMs) the Weyl points (WPs) always appear in pairs in the momentum space with no exception. As a consequence, Fermi arcs occur on surfaces which connect the projections of the WPs with opposite chiral charges. However, this situation can be circumvented in the case of unpaired WP, without relevant surface Fermi arc connecting its surface projection, appearing singularly, while its Berry curvature field is absorbed by nontrivial charged nodal walls. Here, combining angle-resolved photoemission spectroscopy with density functional theory calculations, we show experimentally that a singular Weyl point emerges in PtGa at the center of the Brillouin zone (BZ), which is surrounded by closed Weyl nodal walls located at the BZ boundaries and there is no Fermi arc connecting its surface projection. Our results reveal that nontrivial band crossings of different dimensionalities can emerge concomitantly in condensed matter, while their coexistence ensures the net topological charge of different dimensional topological objects to be zero. Our observation extends the applicable range of the original Nielsen-Ninomiya no-go theorem which was derived from zero dimensional paired WPs with opposite chirality.

[Pay close attention to personnel training in molecular laboratories for hematological neoplasms].

There has consistently growing value of molecular detection to deliver clinical results for diagnosis, classification, prognosis, curative effect prediction and monitoring minimal residual disease of hematological neoplasms. Currently, some clinical laboratories are adopting hematological neoplasms-associated molecular detection in in our country. The increasing demands for clinical testing require that the particular attention need to be paid to the building of the team of professional staff. At present, it is vital to cultivate two kinds of specialized personnel: the technical personnel who are educated in clinical medicine and skilled in developing molecular tests; and the trained managers who are adept at quality control for molecular testing.

[Transcriptome sequencing-based classification and quantification of IKZF1 transcript isoforms in B-cell acute lymphoblastic leukemia].

Objective: To classify and quantify IKZF1 mutant transcripts in B-cell acute lymphoblastic leukemia (B-ALL) by RNA sequencing (RNA-seq) and bioinformatics analysis. Methods: A cohort of 263 B-ALL cases was enrolled at Hebei Yanda Ludaopei Hospital from September 2018 to September 2020. An integrated bioinformatics pipeline was developed to adapt the classification and quantification of IKZF1 transcripts from RNA-seq and was applied to sequencing data of these cases. The IKZF1 mutant transcripts classified by RNA-seq analysis were compared with the qualitative reverse transcription PCR (RT-PCR). Results: IKZF1 mutant transcripts were identified in 53 B-ALL patients by RT-PCR and Sanger sequencing, among which IK6 and IK10 transcripts accounted for 67.9% (36/53) and 28.3% (15/53) respectively. Additionally, 2 patients were double positive for IK6 and IK10. RNA-seq analysis identified 51 patients with IKZF1 mutant transcripts. Compared with the RT-PCR result, the detection sensitivity and specificity of RNA-seq analysis reached 94.3% (50/53) and 99.5% (209/210), respectively. Among the 50 patients with IKZF1 mutant transcripts both in RNA-seq and RT-PCR analysis, the ratio of mutant transcripts to total IKZF1 transcripts in 6 patients was 0.14 (0.11, 0.35), which was significantly lower than that of the other 44 patients [0.88 (0.35, 0.97), Z=-3.945,P<0.001]. IKZF1 mutations mostly occurred in Ph+and Ph-like B-ALL, characterized by abnormal JAK-STAT pathway, and B-ALL with PAX5 translocation. Conclusions: Through the optimized bioinformatics analysis process, RNA-seq data can be used to classify and quantitatively analyze IKZF1 transcripts in B-ALL. Furthermore, the relative expression of mutant IKZF1 transcripts was found to cluster into two groups, and IKZF1 mutation was found often accompanied with PAX5 translocations.

Unravelling the role of arbuscular mycorrhizal fungi in mitigating the oxidative burst of plants under drought stress.

With continued climate changes, soil drought stress has become the main limiting factor for crop growth in arid and semi-arid regions. A typical characteristic of drought stress is the burst of reactive oxygen species (ROS), causing oxidative damage. Plant-associated microbes, such as arbuscular mycorrhizal fungi (AMF), can regulate physiological and molecular responses to tolerate drought stress, and they have a strong ability to cope with drought-induced oxidative damage via enhanced antioxidant defence systems. AMF produce a limited oxidative burst in the arbuscule-containing root cortical cells. Similar to plants, AMF modulate a fungal network in enzymatic (e.g. GmarCuZnSOD and GintSOD1) and non-enzymatic (e.g. GintMT1, GinPDX1 and GintGRX1) antioxidant defence systems to scavenge ROS. Plants also respond to mycorrhization to enhance stress tolerance via metabolites and the induction of genes. The present review provides an overview of the network of plant - arbuscular mycorrhizal fungus dialogue in mitigating oxidative stress. Future studies should involve identifying genes and transcription factors from both AMF and host plants in response to drought stress, and utilize transcriptomics, proteomics and metabolomics to clarify a clear dialogue mechanism between plants and AMF in mitigating oxidative burst.

[ABO gene subtypes and gene expression analysis in three cases of hematological malignancies patients].

Objective: To explore the application and discovery of genotyping, gene sequencing, and gene expression analysis in the determination of ABO blood group subtypes and antigen expression abnormalities in hematological malignancies patients. Methods: From June 2019 to May 2020, three clinical cases were found with forward and reverse ABO typing discrepancy or atypical serologic agglutination pattern in the laboratory and blood transfusion department of Hebei Yanda Ludaopei Hospital were selected. Sequence-specific primer PCR (PCR-SSP) and Sanger sequencing of ABO gene coding regions were performed to determine the ABO genotypes, and whole transcriptome sequencing was used to analyze ABO and FUT1 gene expression levels. Results: A 12-year-old female acute lymphoblastic leukemia patient was determined as O.01.02 and BA.04 sub-genotype, corresponding to the serological B(A) subtype, and her ABO gene expression was normal (354.80). A 41-year-old female acute myeloid leukemia patient was determined as A1.02 and B.01 genotype, corresponding to the serological A(1)B phenotype, and her ABO gene expression was significantly reduced (45.70). A 42-year-old male with myelodysplastic syndrome and myelofibrosis was determined as A1.02 and A2.05 sub-genotype, corresponding to the serological A(1) and A(2) phenotype, respectively, and his ABO expression was negative. FUT1 expression was in the normal range in all three cases. The clinical blood product infusion strategy was formulated according to the genotype and the corresponding immunological subtype, and no significant transfusion-related adverse reactions occurred. Conclusion: Blood group sub-genotypes or aberrant gene expression can lead to ambiguities in serological blood group determination in hematological malignancies patients. ABO genotyping and gene expression analysis can help in this scenario and escort blood product infusion safety.

Observation of Weyl Nodes in Robust Type-II Weyl Semimetal WP_{2}.

Distinct to type-I Weyl semimetals (WSMs) that host quasiparticles described by the Weyl equation, the energy dispersion of quasiparticles in type-II WSMs violates Lorentz invariance and the Weyl cones in the momentum space are tilted. Since it was proposed that type-II Weyl fermions could emerge from (W,Mo)Te_{2} and (W,Mo)P_{2} families of materials, a large number of experiments have been dedicated to unveiling the possible manifestation of type-II WSMs, e.g., surface-state Fermi arcs. However, the interpretations of the experimental results are very controversial. Here, using angle-resolved photoemission spectroscopy supported by the first-principles calculations, we probe the tilted Weyl cone bands in the bulk electronic structure of WP_{2} directly, which are at the origin of Fermi arcs at the surfaces and transport properties related to the chiral anomaly in type-II WSMs. Our results ascertain that, due to the spin-orbit coupling, the Weyl nodes originate from the splitting of fourfold degenerate band-crossing points with Chern numbers C=±2 induced by the crystal symmetries of WP_{2}, which is unique among all the discovered WSMs. Our finding also provides a guiding line to observe the chiral anomaly that could manifest in novel transport properties.

[Trend and characteristics of fall in elderly adults based on data from national injury surveillance sentinel hospitals in Anhui province, 2006-2014].

OBJECTIVE: To understand the incidence pattern of fall among older adults in Anhui province during 2006-2014, and provide scientific basis for intervention strategies and decision-making. METHODS: The incidence data of fall in elderly adults from 6 sentinel hospitals in 2 national injury surveillance areas in Anhui province between 2006 and 2014 were collected for this descriptive epidemiological analysis of the overall incidence trend, demographic characteristics, case distribution and clinical characteristics of fall cases in the elderly. RESULTS: The fall case number, the proportions of fall cases to injury cases and to overall fall cases in the elderly all increased from 2006 to 2014. The fall was the first cause of the elderly injury during the past 9 years. The sex ratio was 0.74 and the cases in males decreased with age. In both males and females, the cases in retirees and the jobless accounted for the highest proportions. The annual incidence peak was during August to October, and two daily incidence peaks were during 9 am-11 am and 16 pm-18 pm. The elderly falls mainly occurred at home(62.07%), and more females were affected than males(70.27% vs. 50.97%). Leisure activity and homework related falls accounted for 49.18% and 28.67% respectively, and more housework related falls occurred in females than in males. Contusion/abrasion was the first injury caused by fall in males(42.63%)and fracture was the first injury caused by fall in females(47.27%). Head was the first injury site in males(38.04%)and leg was the first injury site in females(29.29%). Most injuries caused by fall were mild(55.12%)and moderate(41.84%)in severity, but the proportions of moderate and severe cases increased gradually with age. CONCLUSION: Fall in the elderly has become a public health problem. It is necessary to take targeted prevention and control measures according to the gender and age distributions of the fall in the elderly.

Knockdown of porcine endogenous retroviruses by RNA interference in Chinese experimental miniature pig fibroblasts.

BACKGROUND: The clinical application of porcine-derived xenotransplants is limited by the potential risk of infection due to the presence of porcine endogenous retrovirus (PERV) in tissues, organs, and cells. The establishment of pig fibroblasts with low PERV expression and without PERV-C can provide a nuclear donor to generate a safer transgenic pig. METHODS: In this study, we obtained Chinese Experimental Miniature Pig fibroblasts (CEMPF) with low expression of PERV and none of PERV-C. We designed small interfering RNA (siRNA) expressed as short hairpin RNAs (shRNA) based on the highly conserved gag and pol regions of PERV and screened for the most effective siRNA to inhibit PERV expression. The selected shRNA-pol3 fragment was introduced into the CEMPF to obtain an engineered CEMPF stably expressing shRNA-pol3. RESULTS: The PERV mRNA expression level in the engineered CEMPF was only 7.9% of that observed in fibroblasts from wild-type CEMPF, PERV P15E protein expression was significantly reduced. HEK293 cells cocultured with the supernate of the engineered CEMPF showed no PERV infection. CONCLUSIONS: Engineered CEMPF, which possess no risk of PERV-A/C infection, can serve as a nuclear donor to generate xenograft donor pigs.

Genetic dissection of silicon uptake ability in rice (Oryza sativa L.).

The adequate presence of silicon (Si) in rice plants can enhance their yield and improve their tolerance to various biotic and abiotic stresses. In this study Si uptake abilities were compared between the japonica rice cultivar (cv.) Kinmaze and the indica rice cv. DV85 under three Si concentrations (0.16, 0.4, and 1.6mM) at different time points from 1 to 12h. The results showed that the phenotypic values of two traits-Si uptake by individual plants (SP, Si uptake by all roots of a plant) and Si uptake per unit root dry weight (SR=SP/root dry weight)-of Kinmaze were significantly higher than those of DV85 (P<0.01). Meanwhile, a kinetic study indicated that the Si transporters in Kinmaze and DV85 had the same affinity for silicic acid, but with different Vmax values, indicating that Kinmaze had more Si transporters in the roots than DV85. This may be the main reason for the difference in Si uptake ability between Kinmaze and DV85. In addition, a mapping population consisting of 81 recombinant inbred lines (RILs) derived from the cross between Kinmaze and DV85 was used to detect quantitative trait loci (QTLs) underlying SP and SR. The RILs follow a continuous one-peak distribution and show transgressive segregation in both directions for SP, SR, and root dry weight (RDW). Three QTLs for SP, four for SR, and three for RDW were detected. This can explain 7.16-17.15% of the phenotypic variation (PVE). Thus, the results obtained in this study provide a better understanding of the mechanism of rice Si uptake ability and the basis for fine-mapping the genes involved.

Chromosome variation in dividing protoplasts and cell suspensions of wheat.

The cytology of bread wheat (Triticum aestivum) suspension lines, recycled lines (selected for high division frequency) and their dividing protoplasts, have been examined. Extensive numerical and structural chromosome variation was present in all the lines. The most frequently observed chromosome numbers were around 2n=32, indicating that considerable chromosome loss from the normal wheat complement (2n=6x=42) had occurred during selection of the lines. Chromosome aberrations also indicated loss of chromosome arms and chromosome segments. The implications of this variation for studies on transformation and for the potential regeneration of whole plants from protoplasts of bread wheat are discussed.