BACKGROUND: In China, lung cancer remains the cancer with the highest incidence and mortality rate. Among early-stage lung adenocarcinomas (LUAD), the micropapillary (MPP) component is prevalent and typically exhibits high aggressiveness, significantly correlating with early metastasis, lymphatic infiltration, and reduced five-year survival rates. Therefore, the study is to explore the similarities and differences between MPP and non-micropapillary (non-MPP) components in malignant pulmonary nodules characterized by GGOs in early-stage LUAD, identify unique mutational features of the MPP component and analyze the relationship between the ZNF469 gene, a member of the zinc-finger protein family, and the prognosis of early-stage LUAD, as well as its correlation with immune infiltration. METHODS: A total of 31 malignant pulmonary nodules of LUAD were collected and dissected into paired MPP and non-MPP components using microdissection. Whole-exome sequencing (WES) was performed on the components of early-stage malignant pulmonary nodules. Mutational signatures analysis was conducted using R packages such as maftools, Nonnegative Matrix Factorization (NMF), and Sigminer to unveil the genomic mutational characteristics unique to MPP components in invasive LUAD compared to other tumor tissues. Furthermore, we explored the expression of the ZNF469 gene in LUAD using The Cancer Genome Atlas (TCGA) database to investigate its potential association with the prognosis. We also investigated gene interaction networks and signaling pathways related to ZNF469 in LUAD using the GeneMANIA database and conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Lastly, we analyzed the correlation between ZNF469 gene expression and levels of immune cell infiltration in LUAD using the TIMER and TISIDB databases. RESULTS: MPP components exhibited a higher number of genomic variations, particularly the 13th COSMIC (Catalogue of Somatic Mutations in Cancer) mutational signature characterized by the activity of the cytidine deaminase APOBEC family, which was unique to MPP components compared to non-MPP components in tumor tissues. This suggests the potential involvement of APOBEC in the progression of MPP components in early-stage LUAD. Additionally, MPP samples with high similarity to APOBEC signature displayed a higher tumor mutational burden (TMB), indicating that these patients may be more likely to benefit from immunotherapy. The expression of ZNF469 was significantly upregulated in LUAD compared to normal tissue, and was associated with poor prognosis in LUAD patients (P<0.05). Gene interaction network analysis and GO/KEGG enrichment analysis revealed that COL6A1, COL1A1, COL1A2, TGFB2, MMP2, COL8A2 and C2CD4C interacted with ZNF469 and were mainly involved in encoding collagen proteins and participating in the constitution of extracellular matrix. ZNF469 expression was positively correlated with immune cell infiltration in LUAD (P<0.05). CONCLUSIONS The study has unveiled distinctive mutational signatures in the MPP components of early-stage invasive LUAD in the Asian population. Furthermore, we have identified that the elevated expression of mutated ZNF469 impacts the prognosis and immune infiltration in LUAD, suggesting its potential as a diagnostic and prognostic biomarker in LUAD.
Humans ; Lung Neoplasms/genetics* ; Adenocarcinoma of Lung/genetics* ; China ; Prognosis ; Transcription Factors

Objective:To analyze the clinical characteristics and prognosis of patients infected with novel coronavirus Omicron variant in Shanghai, as to provide a reference for epidemic prevention, clinical diagnosis, and treatment.Methods:Altogether 4 264 novel coronavirus Omicron variant-infected patients with positive results of nucleic acid admitted to Shanghai New International Expo Center N3 Mobile Cabin Hospital from April 2 to May 7, 2022, were included. The demographic and baseline clinical characteristics, treatment strategy, prognosis, and different factors affecting the length of hospital stay were analyzed.Results:A total of 4 264 novel coronavirus variant Omicron-infected cases were collected, including 3 111 cases (73.0%) asymptomatic infections and 1 153 cases (27.0%) mild infections. The overall median age was 45 (33, 55) years old with a range from 2 years old to 81 years old. The male to female ratio was 1.37∶1. Altogether 3 305 cases (77.5%) had been vaccinated, of which 3 166 cases completed more than 2 doses. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical manifestations of these infected patients. During the course of the disease, patients with asymptomatic infection were mainly treated with traditional Chinese medicine (TCM, 55.1%) and clinical observation (36.8%), and those with mild infection were mainly treated with TCM (42.2%) or integrated Chinese and Western medicine (30.4%). All patients were cured and discharged. The overall median length of hospital stay and the negative conversion time of nucleic acid were 9 (6, 10) days and 8 (5, 9) days, respectively. Compared with the asymptomatic infected patients, the hospitalization duration and the nucleic acid negative conversion time of the mildly infected patients were slightly longer [days: 10 (8, 11) vs. 9 (5, 10); 8 (6, 10) vs. 7 (4, 9), both P < 0.001]. Multiple linear regression analysis showed that the increasing age and mild infection were associated with longer hospitalization duration, and the treatment of TCM or integrated Chinese and Western medicine was associated with shortened length of hospital stay (all P < 0.05). Conclusions:The current novel coronavirus Omicron variant epidemic in Shanghai mainly caused asymptomatic and mild infections. The young and middle-aged population had a relatively high infection rate. The upper respiratory tract symptoms such as cough and expectoration were the most common clinical symptoms. Elderly and confirmed patients had prolonged hospitalization duration, while for patients receiving TCM treatment, the hospitalization duration was shortened.

Objective: To optimize the formula of bromhexine hydrochloride dry powder inhalations (BH DPIs). Methods: BH DPIs were prepared by freezing-drying combined with an air-jet milling method. Three factors, including the weights of mannitol (X1), leucine (X2) and poloxamer 188 (X3) in the formula were known to be associated with the quality of BH DPIs. A central composite design was used to investigate the effects of the three factors on the response angle (Y1), fine particle fraction (FPF, Y3) and aerody-namic diameter (Y4). Response surface and overlay contour plot were delineated according to the best-fit mathematic models. Opti-mum formula was selected by overlay contour plot. Results: The quantitative relationships between the three factors and the three re-sponses were obtained. The optimal formula was mannitol﹕leucine﹕poloxamer 188 (2. 4: 2. 22: 0. 05) in the excipients. The pre-dicted and observed values of the optimum formula were similar. Conclusion: The multi-objective simultaneous optimization of the for-mula of BH DPIs is achieved by central composite design-response surface methodology.

Objective To observe the myocardial protective effect of dexmedetomidine in patients undergoing open-heart surgery under cardiopulmonary bypass (CPB).Methods 50 patients of open-heart surgery under CPB were randomly divided into two groups equally, namely observation group and control group.Observation group was treated with injection of dexmedetomidine at 0.5 μg/kg for 15 min, and then maintained at 0.4 μg/kgoh.The control group was given equal volume of normal saline.Concentrations of IMA and cTnI were determined before anaesthesia (t0), after 30 minutes of CPB (t1) and after surgery (t2).Results IMA and cTnI concentrations of t1 and t2 in the observation group were significantly lower than those in the control group (P<0.05).Conclusion Dexmedetomidine has obvious protective effect on myocardium, which can reduce open-heart surgery of patients with myocardial ischemia reperfusion injury (MIRI).

Objective:To explore the effects and mechanism of liraglutide on nitrogen monoxide (NO) release in human umbilical vein endothelial cells in the state of hypoxia and high glucose.Methods:A model of hypoxia and high glucose was established by using isolation and culture of primary human umbilical vein endothelial cells (HUVECs) in vitro.HUVECs were incubated with liraglutide and/or exendin (9-39) for 4 h.The metabolic ability of cells was detected by MTT assay,the activity of lactate dehydrogenase (LDH) was measured by a colorimetric method,and the levels of extracellular NO were measured by a nitrate reductive enzymatic method.The endothelial nitric oxide synthase (eNOS) mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR).Results:Compared with the model group,liraglutide could significantly increase cell metabolic ability,reduce LDH release,increase NO release and eNOS mRNA expression (P<0.05 or P<0.01).The above effects of liraglutide were partly inhibited by glucagon like peptide-1 (GLP-1) receptor antagonist exendin (9-39)(P<0.05).Conclusion:Liraglutide can improve endothelial relaxation function in HUVECs in the state of hypoxia and high glucose in vitro.The effect might be related to up-regulating eNOS mRNA expression and promoting NO release.

Objective To evaluate effects of 4-hydroxyphenyl retinamide (4-HPR) in different vehicles on the proliferation and apoptosis of human keloid fibroblasts (HKFs).Methods A film-ultrasonic dispersion method was used to prepare 4-HPR liposome solution and 4-HPR microbubbles.Primary HKFs were in vitro treated with the 4-HPR liposome solution at different concentrations of 0-80 mg/L for 6-48 hours,and the proliferative activity of HKFs was evaluated by methyl thiazolyl tetrazolium (MTT) assay.Some other HKFs were divided into 3 experimental groups to be treated with 15 mg/L 4-HPR solution (4-HPR solution group),15 mg/L 4-HPR liposome solution (4-HPR liposome solution group) and 15 mg/L 4-HPR microbubbles (4-HPR microbubble group),respectively,and each group was divided into ultrasonic-treated and-untreated subgroups.HKFs without treatment served as control group.After 24-hour treatment,MTT assay was conducted to evaluate the proliferative activity of HKFs in the above groups,flow cytometry to detect apoptosis of HKFs in all groups except the 4-HPR solution group.Results The 4-HPR liposome solution and 4-HPR microbubbles were successfully prepared.MTT assay showed inhibitory effects of 4-HPR liposome solution at concentrations of 1-80 mg/L on the proliferation of HKFs,and the proliferation inhibition rate was positively associated with the drug concentrations (r =0.633,P ＜ 0.01).After the ultrasonic treatment,inhibitory effects on the proliferation of HKFs significantly differed among the 4-HPR microbubble group,4-HPR solution group and 4-HPR liposome solution group (P ＜ 0.01).The 4-HPR liposome solution group and the 4-HPR microbubble group both showed significantly increased apoptosis rates (21.81％ ± 3.73％,39.79％ ± 1.61％,respectively) compared with the control group (6.18％ ± 0.61％,both P ＜ 0.01).Conclusion The 4-HPR microbubbles are successfully prepared,and 4-HPR in different vehicles all can promote HKF apoptosis and suppress HKF proliferation,among which,4-HPR microbubbles in combination with ultrasonic treatment have stronger inhibitory effects than the 4-HPR liposome solution.

Objective:To develop an HPLC method for determining the total ginkgo flavonoid in self-emulsifying drug delivery sys-tem. Methods:Effective chromatographic separation was achieved using a phenomenex C18 column (250 mm × 4. 6 mm, 5 μm) with a mobile phase composed of methanol and water (0.4% phosphoric acid) with the ratio of 50 ∶50 (v/v). The mobile phase was pumped using an isocratic HPLC system at a flow rate of 1. 0 ml·min-1 , the detection wavelength was 360 nm and the column temper-ature was 30 ℃. Results:The three components in the total ginkgo flavonoid were well separated by the proposed method. The linear relationship between the peak area and the concentration was promising within the range of 2. 0-40. 0 μg·ml-1 for quercetin, 3. 0-60. 0 μg·ml-1 for kaempferide and 2. 0-40. 0 μg·ml-1 for isorhamnetin. The mean recovery of quercetin, kaempferide and isorham-netin was 98. 4%, 99. 7% and 100. 5% with RSD of 0. 92%,0. 62% and 1. 24% (n=9), respectively. Conclusion:The method is specific and stable in the determination of total ginkgo flavonoid in self-emulsifying drug delivery system.

Objective To explore the clinicopathological characters of T2 gastric cancer and the value of MSCT in the preop‐erative TNM staging of T2 gastric cancer .Methods A total of 93 patients with T2 gastric cancer were included in our study and un‐derwent preoperative MSCT staging ,who were confirmed by pathologic results .Then the results were compared with those of path‐ologic TNM staging .Also the clinicopathological features of the T2 gastric cancer were analyzed .Results There were no statistical‐ly significant differences in the clinicopathological characters among T2a and T2b patients (P>0 .05) .Comparing with pathologic TNM stage ,the T staging accuracy of MSCT was 91 .40% (85/93) ,the N staging accuracies of CT was 66 .67% (62/93) ,in which , 68 .18% (30/44) ,65 .00% (26/40) ,60 .00% (3/5) and 75 .00% (3/4) were for pN0 ,pN1 ,pN2 and pN3 .And the TNM staging ac‐curacies of CT was 67 .74% (63/93) ,in which ,68 .18% (30/44) ,64 .10% (25/39) ,60 .00% (3/5) and 100% (5/5) were for stageⅠ ,Ⅱ ,Ⅲ and Ⅳ .Conclusion There are no significant different on clinicopathology features among T2a and T2b patients .MSCT can clearly determine the preoperative TNM staging of T2 gastric cancer .

Objective:To investigate the Migration ability toward human pancreatic carcinoma cell line and human colon carcinoma cell line with difference HSP 70 plasma membrane expression .Methods: CD3-CD56+NK cells were obtained from human peripheral blood mononuclear(PBMC)in stem cell growth medium SCGM,2μg/ml TKD was added to the medium on 10th day,the ac-tivating receptor CD94/NKG2C expression levels on NK cells was detected with FAC after 4 days.The human pancreatic carcinoma cell line Colo357 and the human colon carcinoma cell line CW 2 were separated into Colo+and CW2+with high HSP70 expression and Colo-and CW2-with low HSP70 expression;Migration assays of NK to the four difference cell lines were performed in a Transwell cell culture system.The cytolytic activity of TKD-activated NK cells against the four subline with HSP 70 expression on their cell surface was analyzed by MTT assay.Results:Flow cytometry analysis showed that CD 3-CD56+NK cells could expanded after 2 weeks in SCGM medium,and the largest percentage of NK cell was (92.50 ±1.25 )%.CD94 expression levels on NK cells increased obviously after TKD inducement the cell surface HSP 70 expression of Colo+, Colo-were ( 78.2 ±2.2 )% and ( 27.3 ±1.2 )% separately , the cell surface HSP70 expression of CW2+,CW2-were (91.1±2.5)%and (18.2±1.0)%separately after FACS;the Migration of NK cells toward Colo+was (68.6±2.8)%,higher than the migration toward Colo-with (22.8±1.5)%;the Migration of NK cells toward CW2+was(73.5±2.7)%,higher than the migration toward CW2-with (18.2±1.3)%;the cytolytic activity of NK against Colo +was(61.2± 3.0)%compared to (24.5 ±1.5)%against Colo-when the ratio of effector cells and target cell was 20 ∶1,the cytolytic activity of NK against CW2+was (63.8±3.2)%compared to (22.4±1.8)% against CW2-when the ratio of effector cells and target cell was 20∶1.Conclusion:TKD-activated NK cells are highly efficient cytolytic effector cells which have stronger significant migration toward HSP70-positive tumor target cells on their cell surface in vitro .

Rapid detection and classification of bacteria colonies ( Escherichia coli, Listeria monocytogens and Staphylococcus aureus) were investigated by using hyperspectral imaging. The hyperspectral reflectance images (390-1040 nm ) of bacterial colonies on agar plates were collected. Bacterial spectra were extracted automatically based on the masks produced by segmenting a band difference image using the OTSU method. Full wavelength and simplified PLS-DA models were established for classification of bacterial colonies. For the full wavelength model, the overall correct classification rate ( OCCR) and confident OCCR for the prediction set were 100% and 95. 9%, respectively. Besides, competitive adaptive reweighted sampling ( CARS), genetic algorithm ( GA ) and least angle regression-least absolute shrinkage and selection operator ( LARS-Lasso) were used to select feature wavelengths for the development of simplified models. Among them, the CARS-model outperformed the other two in terms of precision, stability and classification accuracy with OCCR and confident OCCR of 100% and 98. 0% for the prediction set, respectively. It was demonstrated that hyperspectral imaging was an effective technology for nondestructive detection of bacterial colonies with high accuracy and high speed. The allocated feature wavelengths by CARS could lay theoretical basis for developing low cost multispectral imaging systems for bacterial colony detection.