Objective To optimize scanning parameters of SPECIAL 1H-MR spectroscopy(MRS),and to observe the feasibility of combining with self-made simulated metabolite spectral data set(B set)of LCModel software for quantitative analysis of pig liver glucose(Glc)in vitro.Methods Metabolite mixture of cod liver oil,Glc and choline with different concentrations of Glc(10,20,30,40,50 mmol/L)and the fixed concentration of cod liver oil(0.125 mg/ml)and choline(100 mmol/L)were prepared with saline to simulate liver metabolism phantoms.There were 5 tube models for each Glc concentration,with 25 tube models configured.SPECIAL sequence was used to scan tube models with different parameters,and 1H-MRS images were obtained.The acquired SPECIAL data of tube models were analyzed using LCModel software and built-in metabolite basic set(A set)and B set,respectively.SPECIAL 1H-MRS images were generated,and signal-to-noise ratio(SNR),standard deviation％(SD％)and Glc signal intensity were obtained.The quality of SPECIAL 1H-MRS images were evaluated according to SNR and SD％,and the optimal scanning parameters were selected.The correlation of Glc signal intensity of phantoms obtained by combining optimal parameters with B set and Glc concentration were analyzed.The optimal SPECIAL sequence was used to scan pig liver in vitro(n=5).Then updated fat suppression(FS)-SPECIAL sequence scanning were performed,the corresponding 1H-MRS images were obtained based on B set,and the quality was observed.Results The optimal scanning parameters of SPECIAL sequence included TR 3 500 ms,TE 4.42 ms,TM 20.00 ms,and the number of repetitions(averages)was 256.SNR of phantoms SPECIAL 1H-MRS acquired with the optimal scanning parameters and B set was 40.5±1.1 and SD％was(13.5±1.0)％,with clearer spectral lines,smoother baselines and higher Glc peak resolution.There was positive correlation between Glc signal intensity obtained with 1H-MRS and Glc concentration of phantoms(r=0.997,P＜0.001).SNR of SPECIAL 1H-MRS of pig liver in vitro was 24.0±2.7 and SD％was(13.5±1.1)％,while SNR of FS-SPECIAL 1H-MRS was 29.5±2.3 and SD％was(4.0±0.8)％,the methylene peak was suppressed and the resolution of Glc peak was higher.Conclusion SPECIAL 1H-MRS with optimized parameters combining with self-made simulation data set of LCModel software could be used for accurately quantitative analysis of pig liver Glc in vitro.

Objective To optimize scanning parameters of SPECIAL 1H-MR spectroscopy(MRS),and to observe the feasibility of combining with self-made simulated metabolite spectral data set(B set)of LCModel software for quantitative analysis of pig liver glucose(Glc)in vitro.Methods Metabolite mixture of cod liver oil,Glc and choline with different concentrations of Glc(10,20,30,40,50 mmol/L)and the fixed concentration of cod liver oil(0.125 mg/ml)and choline(100 mmol/L)were prepared with saline to simulate liver metabolism phantoms.There were 5 tube models for each Glc concentration,with 25 tube models configured.SPECIAL sequence was used to scan tube models with different parameters,and 1H-MRS images were obtained.The acquired SPECIAL data of tube models were analyzed using LCModel software and built-in metabolite basic set(A set)and B set,respectively.SPECIAL 1H-MRS images were generated,and signal-to-noise ratio(SNR),standard deviation％(SD％)and Glc signal intensity were obtained.The quality of SPECIAL 1H-MRS images were evaluated according to SNR and SD％,and the optimal scanning parameters were selected.The correlation of Glc signal intensity of phantoms obtained by combining optimal parameters with B set and Glc concentration were analyzed.The optimal SPECIAL sequence was used to scan pig liver in vitro(n=5).Then updated fat suppression(FS)-SPECIAL sequence scanning were performed,the corresponding 1H-MRS images were obtained based on B set,and the quality was observed.Results The optimal scanning parameters of SPECIAL sequence included TR 3 500 ms,TE 4.42 ms,TM 20.00 ms,and the number of repetitions(averages)was 256.SNR of phantoms SPECIAL 1H-MRS acquired with the optimal scanning parameters and B set was 40.5±1.1 and SD％was(13.5±1.0)％,with clearer spectral lines,smoother baselines and higher Glc peak resolution.There was positive correlation between Glc signal intensity obtained with 1H-MRS and Glc concentration of phantoms(r=0.997,P＜0.001).SNR of SPECIAL 1H-MRS of pig liver in vitro was 24.0±2.7 and SD％was(13.5±1.1)％,while SNR of FS-SPECIAL 1H-MRS was 29.5±2.3 and SD％was(4.0±0.8)％,the methylene peak was suppressed and the resolution of Glc peak was higher.Conclusion SPECIAL 1H-MRS with optimized parameters combining with self-made simulation data set of LCModel software could be used for accurately quantitative analysis of pig liver Glc in vitro.

A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.

The purpose of this study was to investigate the damage mechanism of pathogenic E.coli on mouse brain microvascular endothelial cells(BMEC cells)and mouse alveolar macrophages(MH-S cells),as well as the lung and brain of healthy mice.In this study,BMEC cells and MH-S cells were infected with pathogenic E.coli strains,and cell morphological changes were observed.Plate counting method was used to detect the adhesion and invasion ability of the strains to cells and the number of bacteria in the lungs and brains of mice.RT-qPCR was used to detect the ex-pression of TNF-α,IL-1β and IL-6 genes in cells and mouse organs at different time periods.West-ern blot was used to detect the expression of p-NF-κB,p-JAK2 and p-STAT3 proteins related to inflammation in cells and mouse organs after infection.The results showed that the cell culture medium of the infection group was turbid,the cell vision became dark and blurred,some cells shrank and died,and more fragments were produced.The adhesion rate and invasion rate of BMEC cells at 3 h were significantly lower than those at 6 h(P＜0.050),and the adhesion rate and inva-sion rate of MH-S cells at 3 h were significantly higher than those at 6 h(P＜0.010).Infected mice had a large area of swelling and bleeding in the brain,and the lungs had different degrees of swell-ing and bleeding.The bacterial load in the brain and lung was the highest at 12 h.Compared with the control group,the mRNA expression levels of IL-1β,IL-6 and TNF-α in the infection group were significantly increased at 3 h and 6 h(P＜0.050),and the mRNA expression levels of inflam-matory factors in BMEC cells and MH-S cells were the highest at 6 and 3 h,respectively.The mR-NA expression of inflammatory factors in the brain and lung of infected mice showed a trend of in-creasing first and then decreasing with time,with the highest expression at 12 h after infection.The expression levels of p-NF-κB protein in BMEC cells,MH-S cells,lung and brain tissues of mice in the infection group were significantly higher than those in the control group(P＜0.001),and the expression levels of p-JAK2 protein and p-STAT3 protein were significantly lower than those in the control group(P＜0.050).The above results showed that pathogenic E.coli could adhere and invade BMEC cells and MH-S cells,colonize in lung and brain tissues of mice,promote the expres-sion of NF-κB protein in cells and tissues,inhibit the expression of JAK2 protein and STAT3 pro-tein,and then stimulate cells and tissues to produce inflammatory response.

Objective To compare genomic copy number variations (CNVs) among different subtypes of breast cancer and analyze specific CNVs in each subtype. Methods AIMS software was used for genotype breast cancer (BasL, Her2, LumA and LumB), and GISTIC2.0 software was used to analyze genome-wide CNVs in tumor tissues from TCGA. We collected and analyzed the information and samples of 324 cases of invasive breast cancer admitted to Jiangmen Central Hospital(JMCH). Fluorescence quantitative PCR was used to detect the CNV of ERBB2, TFDP1, MIR148B, CCND1, MDM2 and MIR139 genes in the tumor tissues, to verify TCGA analysis. Results 13q34 was specifically amplified and 12q13.13 was specifically deleted in BasL-type breast cancer. The 17q12 was specifically amplified in Her2 type. The 12q15 was specifically amplified and 11q13.4 was specifically deleted in LumB type, but LumA type had no specific amplification or deletion of chromosome region. The proportion of TFDP1 amplification or MIR148B deletion in BasL type were 57.8% (TCGA) and 71.4% (JMCH), which were significantly higher than other subtypes (P < 0.001). The proportion of ERBB2 amplification of Her2 type were 55.2% (TCGA) and 86.7% (JMCH), which were significantly higher than other subtypes (P < 0.001). The proportion of LumB type with CCND1 or MDM2 amplification or MIR139 deletion were 47.6% (TCGA) and 61.8% (JMCH), which were significantly higher than other subtypes (P < 0.05). Conclusion At the CNV level of breast cancer, Her2 type is characterized by the amplification of ERBB2, BasL type is characterized by the amplification of TFDP1 or the deletion of MIR148B, LumB type is characterized by the amplification of CCND1 or MDM2 or the deletion of MIR139, but LumA type lacks specific CNV characteristics.

Objective To observe the radiosensitization effect of Aidi injection on human lung adenocarcinoma A549 cells, and to analyze its possible mechanism.Methods ① A549 cells were treated with different concentrations of Aidi injection (1.875, 3.75, 7.5, 15, 30, 60 mg/mL) for 24 hours, and in the mean time, a blank control group was set up; the effect of Aidi injection on lung adenocarcinoma A549 cells proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay, and the 10% cell growth inhibitor concentration (IC10) was calculated. ② The experiments were divided into blank control, Aidi control, radiation and Aidi pretreatment groups. The Aidi control group was incubated for 24 hours by Aidi injection IC10; the radiotherapy group was given X-ray irradiation of 4 Gy followed by incubation for 24 hours; the Aidi pretreatment group was incubated for 24 hours by Aidi injection IC10 and then given X-ray irradiation of 4 Gy; the blank control group received equal volume of normal saline and was incubated for 24 hours. The survival fraction (SF) value was detected by cell colony formation assay; the protein levels of the serine phosphorylation at 139 locus of histone (γ-H2AX protein), the key protein in homologous recombination repair pathway (Rad51 protein) and the cell autophage characteristic protein (LC3 protein) were detected by Western Blot; the formation of autophagosome was observed by transmission electron microscope.Results Aidi injection possessed the suppression of the growth of human lung adenocarcinoma A549 cells, the proliferation of the cells in various Aidi groups was lower than that in the blank control group, with the increase in drug concentration, the A549 cell growth inhibition ratio (IR) was gradually increased, representing a dose dependent manner, and the IC10 was 3.09 mg/mL. Compared with the blank control group, the SF value in Aidi control group was not significantly different [(94.7±3.85)% vs. (100.0±0.00)%,P > 0.05], the SF value in radiation group was decreased [(71.8±5.9)% vs. (100.0±0.0)%,P < 0.05], and in Aidi pretreatment group, the value was further decreased compared with that in radiation group [(51.9±4.7)% vs. (71.8±5.9)%,P < 0.05]. Compared with the blank control group, the expression of γ-H2AX protein in the three treatment groups was significantly increased, the degree of increase in Aidi pretreatment group was the most obvious, and it was significantly higher than that in radiation group (gray value: 1.44±0.11 vs. 0.93±0.09,P < 0.05). But the expression of Rad51 protein was the highest in radiation group, and it was higher than that in Aidi pretreatment group (gray value: 1.37±0.07 vs. 0.78±0.04, P < 0.05). Compared with the blank control group, the LC3Ⅱ/LC3Ⅰ value in Aidi control group, radiation group and Aidi pretreatment group were increased, and the degree of increase in Aidi pretreatment group was the most significant (0.35±0.06, 0.37±0.07, 0.49±0.06 vs. 0.05±0.04, allP < 0.05). Under transmission electron microscope, compared with the blank control group, the autophagosome in all treatment groups was increased to some extent, and the degree of increase in Aidi pretreatment group was the most remarkable.Conclusion Aidi injection has the enhancing effect of radiosensitization on human lung adenocarcinoma A549 cells, and its mechanism is possibly related to the up-regulation of A549 cell autophagy level.