Background and objective Bone is a common site for metastasis in lung adenocarcinoma,but the mechanism behind lung adenocarcinoma bone metastasis is still unclear.And currently,there is a lack of easily traceable and stable lung adenocarcinoma bone metastasis cell models,which limits the research on the mechanism of lung adenocarcinoma bone metastasis.The establishment of human lung adenocarcinoma cell line that are highly metastatic to bone,labeled with green fluorescent proteins(GFP)and fireflies luciferase(LUC),along with transcriptomic characterization,would be beneficial for research on lung adenocarcinoma bone metastasis and provide new experimental methods.Methods The human lung ad-enocarcinoma cell line A549-GFP-LUC was injected into nude mice via the left ventricle to construct a bone metastasis model,and was domesticated in vivo for three consecutive times to obtain the human high bone metastasis lung adenocarcinoma cell line A549-GFP-LUC-BM3;cell counting kit-8(CCK-8),colony formation assay,scratch wound assays,Transwell assay and Western blot were used to compare the proliferation and invasion abilities of A549-GFP-LUC-BM3 with the parental cells.A549-GFP-LUC-BM3 cells and parental cells were further analyzed by transcriptomic sequencing.Results Human high-bone metastatic lung adenocarcinoma cells A549-GFP-LUC-BM3 was successfully established.Compared to parental cells,this cells exhibited a significantly higher incidence of bone metastasis and enhanced in vitro proliferation,migration,and invasion abilities.Transcriptomic sequencing results revealed that the A549-GFP-LUC-BM3 cell line had 2954 differentially expressed genes compared to the parental cells,with 1021 genes up-regulated and 1933 genes down-regulated.Gene Ontology(GO)functional enrichment analysis indicated that the differentially expressed genes were primarily localized in cellular components such as the cell periphery.The molecular functions identified as significantly enriched included signaling receptor activity,cal-cium ion binding,and extracellular matrix structural constituent.Additionally,the biological processes found to be enriched were cell adhesion and biological adhesion.The enrichment analysis conducted using the Kyoto Encyclopedia of Genes and Genomes(KEGG)revealed that the differentially expressed genes were primarily involved in the metabolism of xenobiot-ics by cytochrome P450,retinol metabolism,drug metabolism-cytochrome P450,cell adhesion molecules,steroid hormone biosynthesis,and the nuclear factor kappa B(NF-κB)signaling pathway.Conclusion The highly bone-metastatic human lung adenocarcinoma cell line with GFP and luciferase double labeling was successfully established.The biological behavior and transcriptome sequencing of the cell line suggest that it has a high bone-metastatic potential.

Objective:To analyze the molecular epidemiological characteristics of the Corona virus disease 2019 (COVID-19) cases in Shijiazhuang, which can reveal the origin of the outbreak and provide a scientific basis for COVID-19 prevention and control.Methods:From January 2 to January 8, 2021, a total of 404 samples from 170 COVID-19 cases were collected from the Shijiazhuang Fifth Hospital. The consensus sequence of 2019 novel Coronavirus(2019-nCoV) was obtained through multiplex polymerase chain reaction-based sequencing. The sequences of 170 COVID-19 cases were analyzed by the PANGOLIN, and the data were statistically analyzed by T-test.Results:Among the 404 COVID-19 samples, a total of 356 samples obtained high quality genome sequences (>95%,100×sequencing depth). The whole genome sequences of 170 COVID-19 cases were obtained by eliminating repeated samples. All 170 sequences were recognized as lineage B1.1 using PANGOLIN. The number of single nucleotide polymorphism arrange from 18-22 and most of the single nucleotide polymorphism were synonymous variants. All of 170 genomes could be classified into 48 sub-groups and most of the genomes were classified into 2 sub-groups (66 and 31, respectively).Conclusions:All cases in this study are likely originated from one imported case. The viruses have spread in the community for a long time and have mutated during the community transmission.

Objective To indentify the cognitive status of Chinese patients to acne and the influencing factors to theirs' cognitive status,so as to provide solid evidences for the prevention and treatment of acne.Methods A self-designed questionnaire was made to conduct this survey of 16,156 acne patients,who seeked to the treatment in the dermatological departments from 112 hospitals in China.The survey consisted of several parts,including the general status of patients,the patients' cognition of occurrence,development and risk factors of acne,whether the first choice was seeking treatment at the hospital when the patients had acne and the condition of selection of skin care products.The factors were analyzed,which could impact the cognition of the patients' behavior of treatment,how did the patients' cognition to influence their medical behavior and skin care as well as the consistency of assessment of the severity of acne by doctors and patients themselves.Results The acne patients studied had the best knowledge of "acne is a skin disease","it not only occurs in the period of adolescence" and "the disease can be prevented and cured",which accordingly accounted for 80.65％,69.16％ and 65.49％ of the total patients respectively.However,the awareness of acne patients to heredity,high sugar and dairy products as risk factors for acne was insufficient,which accounted for 48.72％,42.40％ and 18.25％ of the total patients,respectively.Gender,age,educational level,occupation and health knowledge were the main factors affecting the cognitive level of patients;the survey also found that men,patient with educational level of junior high or even lower educational condition,occupation of labor workers or farmers and patients were lack of health education with poor knowledge of the genetics and dietary were risk factors for acne;patients with age over 36 years or with mild illness had poor knowledge of dietary risk factors for acne;the difference was statistically significant (P＜0.05).The analysis of the influence of cognitive status on medical treatment behavior and skin care showed that the better the cognition,the higher the probability of patients would choose medical treatment as the first choice as well as choosing functional skin care products;the difference was statistically significant (P＜0.05).The consistency of assessment of the severity of acne by doctors and patients was poor (Kappa value ＜0.4),and the assessment of severity of acne by patients was more serious than doctors' assessment.Conclusions Patient's cognitive status will affect their medical behavior and skin care,and there is also a phenomenon that patients have a more serious assessment of their acne condition.It is suggested that health education for acne patients should be strengthened in clinical medicine so as to improve their knowledge of acne as well as preventing from acne effectively.

Objective To determine the expression of interleukin-6 (IL-6) in cystic lesions of patients with acne vulgaris,and to evaluate the in vitro effect of Propionibacterium acnes (P.acnes) on the production of IL-6 and activation of p38 mitogen-activated protein kinase (p38MAPK) in the human acute monocytic leukemia cell line THP-1.Methods Real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of IL-6 in cystic lesions of 6 patients with acne vulgaris,as well as in skin tissues of 6 healthy persons.Some cultured THP-1 cells were divided into 5 groups to be treated with 2 × 106 CFU/ml,2 × 107 CFU/ml and 2 × 108 CFU/ml heat-killed P.acnes suspensions (P.acnes groups),100 μμtg/L lipopolysaccharide (LPS group) and RPMI 1640 medium (control group) respectively.After 1-,3-and 6-hour treatment,real-time fluorescence-based quantitative PCR was conducted to determine the mRNA expression of IL-6 in the above groups.Enzyme-linked immunosorbent assay (ELISA) was performed to detect the level of IL-6 in the culture supernatant of cells in the 2 × 108-CFU/ml P.acnes group,LPS group and control group at 24 hours after the treatment.Western blot analysis was conducted to determine the protein expression of p38MAPK and phosphorylated p38MAPK in the 2 × 108-CFU/ml P.acnes group after 15-,30-and 60-minute treatment,as well as in the LPS group after 30-minute treatment and in the control group.Some other THP-1 cells were divided into 3 groups:2 × 108-CFU/ml P.acnes group treated with 2 × 108 CFU/ml P.acnes suspensions,SB203580 (an inhibitor of p38MAPK) group treated with 20 μmol/L SB203580 for 30 minutes followed by the treatment with 2 × 108 CFU/ml P.acnes suspensions,and control group treated with RPMI 1640 medium alone.After 6-hour treatment,the mRNA expression of IL-6 in the above 3 groups was measured by real-time fluorescencebased quantitative PCR.Results The mRNA expression of IL-6 was significantly higher in the cystic lesions of acne vulgaris than in the normal skin tissues (3.680:±:0.790 vs.1.155 ± 0.250,t =3.047,P ＜0.05).Two-way analysis of variance showed that there were significant difference in the mRNA expression of IL-6 among the 2 × 106-CFU/ml,2 × 107-CFU/ml and 2 × 108-CFU/ml p.acnes groups,LPS group and control group (F =532.3,P ＜ 0.001,v =4),and the mRNA expression of IL-6 significantly differed among different time points (F =526.6,P ＜ 0.001,v =2).There were also significant differences in the IL-6 level in the culture supernatant of cells among the 2 × 108-CFU/ml p.acnes group ([1 618.22 ± 32.23] ng/L),LPS group ([3 212.06 ± 353.00] ng/L) and control group ([147.10 ± 0.53] ng/L;v =2,F =102.35,P ＜0.01).After 15-,30-and 60-minute treatment with 2 × 108 CFU/ml P.acnes suspensions,the protein expression of phosphorylated p38MAPK obviously increased.The mRNA expression of IL-6 in THP-1 cells was significantly lower in the SB203580 group than in the 2 × 108-CFU/ml p.acnes group (t =15.91,P =0.004).Conclusions The mRNA expression of IL-6 evidently increases in the cystic lesions of patients with acne vulgaris.P.acnes can activate the signaling molecule p38MAPK in THP-1 cells,and promote the production of IL-6 by THP-1 cells.

Objective To evaluate the effect of Aspergillus fumigatus on the expression of tumor necrosis factor-α (TNF-oα) and activation of intracellular signaling molecule p38 mitogen-activated protein kinase (p38MAPK) in a human acute monocytic leukemia cell line THP-1.Methods Cultured THP-1 cells (2 x 105/ml) were divided into 4 groups to be treated with Aspergillus fumigatus suspensions at concentrations of 106 and 107 colony-forming units (CFU)/ml (106-and 107-CFU/ml Aspergillusfumigatus groups),100 mg/L β-glucan (a positive stimulus,β-glucan group),culture medium (blank control group) respectively for 1,3 and 6 hours.Real-time fluorescence-based quantitative PCR (qPCR) was conducted to determine the mRNA expression of TNF-α in the THP-1 cells in the above groups.Some other THP-1 cells were treated with 107 CFU/ml Aspergillusfumigatus suspensions (107-CFU/ml Aspergillusfumigatus group),β-glucan (β-glucan group) and culture medium (blank control group) separately for 24 hours,and enzymelinked immunosorbent assay (ELISA) was performed to detect the level of TNF-α in the culture supernatant of THP-1 cells.Western blot analysis was conducted to detect the levels of p38MAPK and phosphorylated p38MAPK in THP-1 cells after 15-,30-and 60-minute treatment with 107 CFU/ml Aspergillusfumigatus suspensions.After 2-hour incubation with the p38MAPK inhibitor SB203580 (20 μmol/L),some THP-1 cells were additionally treated with 107 CFU/ml Aspergillus fumigatus suspensions,β-glucan and culture medium separately for 6 hours,and those without SB203580 treatment served as the control group.Then,qPCR was performed to measure the mRNA expression of TNF-α in the THP-1 cells in the above groups.Results The mRNA expression of TNF-α significantly differed among the 106-and 107-CFU/ml Aspergillus fumigatus groups,β-glucan group and blank control group (F =110.983,P ＜ 0.001),and significantly increased over time (F =701.680,P ＜ 0.001).After 24-hour treatment with 107 CFU/ml Aspergillus fumigatus suspensions,the TNF-α level(6 236.30 ± 437.12 ng/L)significantly increased compared with the blank control group (132.10 ± 0.61 ng/L,P ＜ 0.01).Thirty minutes after the treatment with 107 CFU/ml Aspergillusfumigatus suspensions,the phosphorylated p38MAPK level significantly increased,but started to decrease at 60 minutes.The mRNA expression of TNF-α was significantly lower in the SB203580-treated Aspergillusfumigatus groups (3.83 ± 0.62) than in the SB203580-untreated Aspergillus fumigatus groups (187.23 ± 21.62).Conclusion After the treatment with Aspergillus fumigatus,human THP-1 cells can activate the signal molecule p38MAPK and secrete TNF-α,suggesting that monocytes may participate in the innate immune response to Aspergillusfumigatus infection.

Objective To compare the value of automatic breast volume scanner (ABVS) with enhanced magnetic resonance imaging(MRI) in the diagnosis of breast masses.Methods Seventy-four patients with 80 breast masses underwent preoperative ultrasound examinations including ABVS and MRI.The values of ABVS and MRI in the diagnosis of breast masses were comparatively analyzed.Results Among the 80 breast masses that were surgically removed,37 masses were benign and 43 masses were malignant.The sensitivity,specificity,accuracy,positive predictive value and negative predictive value of ABVSin the diagnosis of breast malignant masses were 94.6％,79.1％,86.3％,79.5％ and 94.4％,respectively,those of MRI were 94.6 ％,86.0 ％,90.0 ％,85.4 ％ and 94.9 ％,respectively,and those of the combination of ABVS and MRI were 94.6％,93.0％,93.8％,92.1％,and 95.2％,respectively.The sensitivity and specificity were not significant difference between ABVS and MRI in the diagnosis of breast malignant masses(P ＞0.05).The specificity of the combination ABVS and MRI in the diagnosis of breast malignant masses were significantly higher than that of ABVS (x2 =4.17,P =0.04).The sensitivity,specificity and accuracy of convergence sign in the diagnosis of breast malignant masses were 64.9 ％,97.7 ％ and 82.5 ％,respectively.Conclusions ABVS and MRI are both valuable in the diagnosis of breast masses,and the combination of ABVS and MRI is the most valuable due to high specificity.

Objective To investigate the diagnosis, therapeutic monitoring and prognosis value of the total procollagen type 1 amino-terminal propeptide(tP1NP), beta-C-terminal telopeptide(β-CTx)and bone alkaline phosphatase(BAP)in the bone metastasis of lung cancer.Methods With the case-control study method, the serum levels of tP1NP, β-CTx and BAP in 196 lung cancer patients, including 109 patients with bone metastases,87 patients without bone metastases,and 106 healthy controls at the Shanghai Sixth People′s East Hospital affiliated to Shanghai Jiao Tong University between 2014 and 2015 were quantitatively detected by chemiluminescent immunoassay.Receiver operating characteristic(ROC)curve was calculated to assess the diagnostic value.Survival curve was performed by Kaplan-Meier method. Results The concentration of tP1NP,β-CTx and BAP in the lung cancer patients with bone metastasis were significantly higher than that in the lung cancer patients without bone metastasis(Z=-5.642,P<0.001;Z=-3.783,P<0.01;Z=-8.923,P<0.01).ROC curve analysis showed that the AUC of tP1NP, β-CTx and BAP were 0.874,0.776 and 0.678 respectively(P<0.05).The AUC of the combined three markers was 0.925(95%CI 0.867-0.963),with sensitivity of 77.11% and specificity of 98.11%.The levels of tP1NP and β-CTx were associated with the clinical response.The concentration of tP1NP,β-CTx were significantly decreased in patients achieved remission(t=4.607,P<0.05;t=5.355,P<0.05). Survival analysis showed that higher concentration of tP 1NP was correlated with poor prognosis[OR=3.287, 95%CI(1.118-9.661),P<0.05].Conclusions The levels of tP1NP,β-CTx and BAP cloud be used for the differential diagnosis of bone metastasis of lung cancer,and the combined usage was more effective. tP1NP and β-CTx cloud be used in therapeutic monitoring of lung cancer patients with bone metastasis. Moreover,tP1NP could be used as prognostic biomarker in lung cancer patients.

Background and purpose:Stanniocalcin 1 (STC1) has been reported to be up-regulated in various cancer tissues, and related to malignancy degree of cancer. However, the molecular mechanism of STC1 in lung cancer cells is still not clear. This experiment aimed to investigate the effects of STC1 on cell cycle and apoptosis of lung cancer A549 cells.Methods:A549 cells were transfected with validated siRNA for STC1 A549-STC1-siRNA and a negative control vector RNA A549-Vector. The gene and protein expression of cell cycle-related genes, including CyclinA, CyclinB1, CyclinD1, CyclinE, CDK2 and CDK4, as well as apoptosis-inhibiting genes Bcl-2, Bcl-xl and apoptosis-inducing genes Caspase-3, Bax, Bak and Bid, were detected by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. The cell cycle distribution was determined with lfow cytometry. Terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) was used to detect cell apoptosis.Results:After transfection with STC1-siRNA, the gene and protein expression of CyclinA, CyclinB1, CyclinD1, CyclinE, CDK2 and CDK4 decreased signiifcantly in A549 cells (P<0.05). The proportion of cells in G0/G1 phase signiifcantly increased,
whereas the proportion of cells in S phase and G2/M phase decreased (P<0.05). The cell cycle was blocked at G0/G1 phase. Furthermore, compared with that in A549-Vector, the gene and protein expression of Bcl-2 and Bcl-xl in A549-STC1-siRNA was reduced signiifcantly (P<0.05), while the expression of apoptosis-inducing genes Caspase-3, Bax, Bak and Bid increased obviously (P<0.05). In addition, the percentage of apoptotic cells significantly increased in A549-STC1-siRNA compared with that in A549-Vector detected by TUNEL method.Conclusion:Down-regulation of STC1 by RNAi can block the cell cycle of A549 cells, inhibit cell proliferation, and promote cell apoptosis.

Soil particles are very heterogeneous in microscopic scale, which is manifested the double-layer structure made of the soil organic matter and mineral matter. In this work, Fourier by transform infrared photoacoustic spectroscopy ( FTIR-PAS) combined with independent component analysis ( ICA) was utilized for in situ depth-profiling of the manmade complex film, in order to lay a foundation of in situ characterizing the heterogeneous soil organic-mineral complex. The complex film was composed of the PE preservative film and office adhesive tape. The moving velocity of infrared photoacoustic spectrometer was set to 0. 16 cm/s, 0. 32 cm/s and 0. 64 cm/s, respectively. Independent component analysis ( ICA ) was performed on the photoacoustic spectra of the heterogeneous complex film. Results showed that the depth-resolved information of the complex film could be derived by changing the moving velocity, and the estimated thickness of PE film was 5. 4-7. 6 μm, which was close to the actual thickness 7 ± 1 μm. Moverover, the spectral features of the polyethylene ( PE) preservative film and office adhesive tape were extracted from the photoacoustic spectra of the heterogeneous complex film by means of ICA. Depth profiling of complex film samples showed that FTIR-PAS could be used as a new analytical tool to study heterogeneous soils, especially soil organic-mineral complexes.

In this study, we explored the rationality of processing methods and mechanism of Aconiti Lateralis Radix (Fuzi) through comparing the chemical contents of diester alkaloids (DAs) and monoester alkaloids (MAs) in the raw material of Fuzi and its processed products. The results showed that the toxicity potency of MAs is at least lower than 1/64 to 1/180 of the toxicity potency of DAs. The contents of DAs in processed Fuzi decreased to 1/76.5 to 1/38.3 of the value of raw Fuzi. The contents of MAs in processed Fuzi significantly increased by 4.6 to 5.2 fold or basically the same as that of the raw Fuzi. The values of MAs/DAs of processed Fuzi were enhanced by 30 to 390 fold of the raw Fuzi. It was found that the contents of DAs were insignificantly different between "Wu dan fu pian" (steaming or stir-frying without Danba) and "Dan fu pian" (steaming or stir-frying with Danba). The result suggested that the abilities of "eliminating toxicity" of different processing methods were equivalent at all. In contrast, the contents of MAs contained in "Wu dan fu pian" were of 5.3 to 8.7 fold higher than the values in "Dan fu pian". This result suggested the processing method by steaming or stir-frying without Danba might have better effect for "conserving property" than the method processed with Danba stipulated by China Pharmacopoeia. We believe that the new processing method without Danba can be recommended in further application due to it offers a simple procedure and it will not introduce inorganic impurities in the products.