The protein kinase A/protein kinase G/protein kinase C-family (AGC kinase family) of eukaryotes is involved in regulating numerous biological processes. The 3-phosphoinositide- dependent protein kinase 1 (PDK1), is a conserved serine/threonine kinase in eukaryotes. To understand the roles of PDK1 homologous genes in cell death and immunity in tetraploid Nicotiana tabacum, the previuosly generated transgenic CRISPR/Cas9 lines, in which 5-7 alleles of the 4 homologous PDK1 genes (NtPDK1a/1b/1c/1d homologs) simultaneously knocked out, were used in this study. Our results showed that the hypersensitive response (HR) triggered by transient overexpression of active Pto (Pto^Y207D) or soybean GmMEKK1 was significantly delayed, whereas the resistance to Pseudomonas syrangae pv. tomato DC3000 (Pst DC3000) and tobacco mosaic virus (TMV) was significantly elevated in these partial knockout lines. The elevated resistance to Pst DC3000 and TMV was correlated with the elevated activation of NtMPK6, NtMPK3, and NtMPK4. Taken together, our results indicated that NtPDK1s play a positive role in cell death but a positive role in disease resistance, likely through negative regulation of the MAPK signaling cascade.
Nicotiana/virology* ; Disease Resistance/genetics* ; Plant Diseases/immunology* ; Plants, Genetically Modified/genetics* ; Gene Knockout Techniques ; Plant Proteins/genetics* ; CRISPR-Cas Systems ; Protein Serine-Threonine Kinases/genetics* ; 3-Phosphoinositide-Dependent Protein Kinases/genetics* ; Pyruvate Dehydrogenase Acetyl-Transferring Kinase ; Tobacco Mosaic Virus/pathogenicity*

AIM:To analyze the compliance rate and possible influencing factors for blood concen-tration of meropenem in patients with central ner-vous system(CNS)infections,and to provide basis for optimizing the administration of meropenem in the patients.METHODS:A retrospective analysis was performed in patients who were treated with meropenem and monitored the concentration(Cmin)from January 2021 to December 2022.The basic clinical data of the patients were recorded,and the achievement of meropenem blood concentration was counted.Univariate and binary logistic regres-sion analysis were used to analyze the influencing factors for the e substandard steady-state blood trough concentration of meropenem.The ROC curve was drawn to predict the clinical characteris-tics.RESULTS:Forty-five cases were included.The median Cmin of meropenem was 4.14(0.82,16.29)mg/L,and 51.11％reached the target range with 4 mg/L.Binary logistic regression analysis showed that serum creatinine value and using mannitol were risk factors for the substandard steady-state blood trough concentration of meropenem(P＜0.05).When serum creatinine value was less than 84 μmol/L,the area under the ROC curve was the largest(0.916),the sensitivity was 0.783,the speci-ficity was 0.955,and the Youden index was 0.738.CONCLUSION:The Cmin compliance rate of merope-nem blood concentration in patients with CNS in-fection is low,especially in patients with hyperfunc-tion of kidney and those who using mannitol.It is necessary to conduct drug concentration monitor-ing to optimize the administration of meropenem in patients with CNS infection.

AIM:To investigate the effect of hypoxia-preconditioned human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSC-Exos)on pulmonary vascular endothelial-mesenchymal transition(EndMT)in hypoxic pulmonary hypertension(HPH).METHODS:(1)Primary hUCMSCs were isolated and cultured by tissue adhesion method,and hUCMSC-Exos were extracted by ultrafiltration and identified.(2)Twenty-four SPF male SD rats were ran-domly divided into normoxia(N)group,hypoxia(H)group,hypoxia+normoxic hUCMSC-Exos group and hypoxia+hypoxia-preconditioned hUCMSC-Exos group,with 6 rats in each group.The rats in H group and intervention groups were placed in a cabin that simulated the hypoxic environment at an altitude of 5 000 m,and normoxic hUCMSC-Exos,hypoxia-precon-ditioned hUCMSC-Exos or equivalent volume of PBS were injected through the tail vein on the 3rd,5th,7th,10th and 14th days in hypoxia environment.After 21 d of modeling,the right ventricular systolic pressure(RVSP)and right ven-tricular hypertrophy index(RVHI)of the rats were detected,and the pathological changes of lung tissues were observed by HE staining.(3)After starvation for 12 h,human pulmonary arteriole endothelial cells(HPAECs)were randomly di-vided into normoxic control(N-Con)group,hypoxic model(H-Con)group,hypoxia+normoxic hUCMSC-Exos group and hypoxia+hypoxia-preconditioned hUCMSC-Exos group.The migration ability and tube formation ability of HPAECs were detected by Transwell assay and tube formation experiment.The expression of CD31 and α-smooth muscle actin(α-SMA)in HPAECs was detected by immunofluorescence double-staining.The protein levels of CD31,VE-cadherin,α-SMA and vimentin in pulmonary vessels and HPAECs were assessed by Western blot.RESULTS:(1)The HPH rat model was suc-cessfully established after 21 d of hypoxia,and EndMT occurred in pulmonary vessels.Compared with N group,the levels of RVSP,RVHI,percentage of vascular wall area(WA%)and percentage of vascular wall thickness(WT%)in H group were significantly increased(P＜0.01),pulmonary vascular wall thickening and the protein levels of CD31 and VE-cad-herin were significantly decreased(P＜0.01),while the protein levels of α-SMA and vimentin were significantly increased in pulmonary vessels(P＜0.05 or P＜0.01).Compared with H group,the RVSP,RVHI,WA%and WT%(P＜0.01)were significantly decreased(P＜0.05 or P＜0.01),and pulmonary vascular remodeling was attenuated after normoxic or hypoxia-preconditioned hUCMSC-Exos intervention.After hypoxia-preconditioned hUCMSC-Exos intervention,HPH pul-monary vascular remodeling and EndMT formation were significantly inhibited.(2)After 48 h of hypoxic treatment,the migration,tubule formation and EndMT of HPAECs were induced.Compared with H-Con group,cell migration and tube formation were significantly decreased after hypoxia-preconditioned hUCMSC-Exos intervention(P＜0.01).The protein levels of CD31 and VE-cadherin were increased,while the protein levels of α-SMA and vimentin were decreased(P＜0.05 or P＜0.01).CONCLUSION:Hypoxia-preconditioned hUCMSC-Exos attenuate the formation of HPH pulmonary vascu-lar remodeling by inhibiting pulmonary vascular EndMT.

Due to the virtues of no stutter peaks, low rates of mutation, and short amplicon sizes, insertion/deletion (InDel) polymorphism is an indispensable tool for analyzing degraded DNA samples from crime scenes for human identifications (Wang et al., 2021). Herein, a self-developed panel of 43 InDel loci constructed previously by our group was utilized to evaluate the genetic diversities and explore the genetic background of the Han Chinese from Beijing (HCB) including 301 random healthy individuals. The lengths of amplicons at 43 InDel loci in this panel ranged from 87 to 199 bp, which indicated that the panel could be used as an effective tool to utilize highly degraded DNA samples for human identity testing. The loci in this panel were validated and performed well for forensic degraded DNA samples (Jin et al., 2021). The combined discrimination power (PD) and combined probability of exclusion (PE) values in this panel indicated that the 43 InDel loci could be used as the candidate markers in personal identification and parentage testing of HCB. In addition, population genetic relationships between the HCB and 26 reference populations from five continents based on 19 overlapped InDel loci were displayed by constructing a phylogenetic tree, principal component analysis (PCA), and population genetic structure analysis. The results illustrated that the HCB had closer genetic relationships with the Han populations from Chinese different regions.
Beijing ; China ; Forensic Genetics/methods* ; Gene Frequency ; Genetics, Population ; Humans ; INDEL Mutation ; Phylogeny

Objective : To establish the liver conditional knockout mouse model ofNod2gene infected byKlebsiella pneumoniae(K.pneumoniae),and to explore the role and mechanism ofNod2gene in the process of liver abscess caused byK.pneumoniaeinfections. Methods : Nod2flox/floxmice were obtained by self-crossing ofNod2flox/+mice, andAlb-Cre+mice were hybridized withNod2flox/+to obtainNod2flox/+;Alb-Cre+mice, then the above two genotypes mice were crossed to obtain liver conditional knockout mice ofNod2gene(Nod2flox/flox;Alb-Cre+) and negative control mice in the same litter(Nod2flox/flox).The genomic DNA of mice toe was extracted and amplified by polymerase chain reaction(PCR).The genotypes of offspring were identified by agar-gel electrophoresis and the livers of mice were extracted.Real-time fluorescence quantitative PCR(RT-qPCR) and Western blot were used to verify the knockout efficiency ofNod2gene in the liver.Both experimental group and control group mice were infected withK.pneumoniae,and the survival rate and pathological changes of livers were observed at different time points, and mRNA expression levels of Tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β) and C-X-C motif chemokine ligand 1(CXCL1) in the livers of experimental group and control group were detected by RT-qPCR 24 h postK.pneumoniaeinfections. Results : The expression of NOD2 mRNA in the liver ofNod2flox/flox;Alb-Cre+mice decreased, and the Western blot results showed that the expression of NOD2 protein decreased.Compared with the control group, the survival rate of mice infected withK.pneumoniaein the experimental group decreased(median survival time=60.5 h,P=0.046 9) and the liver tissue showed more serious pathological damage, furthermore the mRNA expression levels of TNF-α,IL-1β and CXCL1 in the livers of experimental group were lower than those of the control group, and the difference was statistically significant(P<0.05). Conclusion NOD2 plays a protective role in the process of liver abscess induced byK.pneumoniaeinfections.

Objective To observe the oral part of the facial artery and facial vein and to provide anatomical data for clinical applica-tion. Methods The origin, branches, course, diameter, position of oral part of facial artery and facial vein were observed on 32 fixed cada-ves (64 sides). Results The position relation between the facial artery and facial vein is non-constant. Measure the distance from inferior border of mandible to corner of the mouth, angulus mandibulae, mental protuberance midpoint. It is (5. 49 ± 0. 63) cm, (2. 50 ± 0. 89) cm and (6. 20 ± 1. 68) cm in the left side respectively, and (5. 69 ± 0. 72) cm, (2. 56 ± 1. 08) cm and (6. 85 ± 1. 86) cm in the right side re-spectively. The diameter of facial artery in inferior border of mandible is (0. 33 ± 0. 08) cm in the left side and (0. 38 ± 0. 07) cm in the right side;while the diameter of facial vein is (0. 40 ± 0. 12) cm in the left side and (0. 42 ± 0. 18) cm in the right side. The facial artery and facial vein are not concomitant and they are not asymmetry also. The position of superior labial artery arteries is constant, but the position of inferior labial artery arteries have more variations. Conclusion The branches, course, diameter and position of oral part of facial artery and facial vein have a number of variations. The superior labial artery arteries could be positioned more easily than inferior labial artery arter-ies. Being familiar with their distribution is of great importance for clinical application.

An 18-year-old female presented with painful erythema and nodules on both legs for more than 2 years.Dermatological examination showed irregularly sized,mildly indurated,tender,deep subcutaneous nodules arising in diffused infiltrated dark erythematous patches in the inner and posterior region of the left leg.Histopathology showed no significant changes in the epidermis.There were perivascular lymphoid cell infiltrates in the dermis and subcutis.Multiple sites of necrosis of blood vessel walls with vascular occlusions were noted.The lumens of some blood vessels were filled with lymphocytes,among which were many atypical cells with hyperchromatic nuclei and pathologic mitotic figures.Immunohistochemistry showed that lymphocytes in the cavities of blood vessels were positive for CD3(+++),CD3ε(+++),CD2(+),CD56(+++),granzyme B(+++),perforin(+++),CD30(+),Ki67 (+++,100％ ),but negative for CD20,CD5,CD7,CD4,CD8,TdT,anaplastic lymphoma kinase,early membrane antigen (EMA) or pan cytokeratin (pCK).The endothelial cells lining the blood vessels stained positively for CD34.The intravascular lymphocytes were also positive for EBER1/2 by in situ hybridization.A diagnosis of cutaneous intravascular NK/T cell lymphoma was made.

0.98); immature cells would display in the WBC histogram when in higher proportion. Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to checked with microscopy for DC before reported.

0.99). DC: reproducibility was good for neutrophils, lymphocytes monocytes, eosinophils and basophils. Comparison of the results by instrument with manual for normal samples in morphology, the correlation was better for neutrophils, lymphocytes and eosinophils (r:0.968～0.983) ,good for monocytes(r=0.917), not good for basophils(r=0.659);The WBC scattergram would change and alarm flags would display when there are neutrophilic stab granulocytes, abnormal or atypical lymphocytes and immature cells in higher proportion.Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to check with microscopy for DC before reported when it were doubtted.

0.05).RBC and haematocrit(HCT)were significantly decreased(P