Objective To explore the influence of autophagy on lipopolysaccharide (LPS) induced acute lung injury (ALI) . Methods Forty‐eight Sprague Dawley (SD) rats were randomly divided into four groups ,12 cases in each group :(1)normal saline control group (NS) ,(2)LPS model group (L) ,(3) LPS and autophagy group (L +A) and (4) LPS and autophagy inhibition group (L+I) .Arterial blood samples was obtained for detecting the blood gas ,including PaO2 ,PaCO2 and pH ,and the lung tissue dry/wet ratio was calculated .The HE staining was used to observe the histopathological changes of lung tissue .Moreover the lung le‐sion score was performed ;the expression of microtubule associated protein ,light chain protein 3b(LC3b) ,myeloperoxidase(MPO) , macrophage inflammatory protein 2(MIP‐2) ,interleukin‐1β(IL‐1β) and tumor necrosis factor‐α(TNF‐α) in serum and bronchoalve‐olar lavage fluid(BALF) was assessed by ELISA .Results Compared with the NS group ,arterial blood PaO2 and pH in the group L were decreased and PaCO2 was increased (P<0 .05);compared with the L group ,the arterial blood PaO2 and pH in the L+A group were increased and PaCO2 was declined (P<0 .01) ,the arterial blood PaO2 and pH in the L+ I group were decreased and PaCO2 was elevated ,the differences were statistically significant (P<0 .01) .The LC3b concentration in serum and BALF in the L group and L+I group was declined ,while MPO ,MIP‐2 ,IL‐1βand TNF‐αconcentrations were increased ,while which in the L+ A group were just the opposite .Conclusion Autophagy plays a improvement and protective effect on LPS induced acute lung injure in rat .

Objective To study the CT signs of bronchotracheal transparent foreign body and its diagnostic value.Methods The X-ray and CT findings of bronchotracheal trasparent foreign body in 46 cases were analysed comparativly.Results Among 46 cases,9 cases of trachea foreign body,13 cases of right bronchial foreign body,7 cases of left bronchial foreign body,2 cases of both sides of bronchial foreign body were showed directly by CT,and 8 cases of right bronchia foreign body,6 cases of left bronchia foreign body,1 case of both sides of bronchia foreign body showed indirectly on CT,the sensitivity was 100 percent,while the foreign body and its anatomical location could not be showed on X-ray film,but the indirect signs just like pulmonary emphysema,obstructive pulmonitis and drowned lung etc,could be showed by X-ray films in 34 cases.In another 12 cases were negative,the sensitivity was 73.9 percent.Conclusion The transparent foreign body can not be showed but indrect signs by X-ray film;While CT scan can not only shows the anatomical location of foreign body,but also the shape and size of the foreign body,it offers more information than X-ray films and has important refering value to clinicians to take out the foreign body.

Objective To discuss the diagnostic value of color Doppler echocardiography in small coronary-to-pulmonary fistula (SCPF) in children. Methods The clinical data of children who were diagnosed CPF by color Doppler echocardiography during 2011-2012 in my hospital were analyzed retrospectively. Results Seventeen cases with CAF including one diagnosed by forensic report and 8 cases diagnosed by AGA. Conclusion The results indicated that SCPF detection rate could be improved largely by observing spraying-up sign experienced in pulmonary artery diastolic combined with rich flow signal of coronary in color Doppler. In addition, it is valuable to diagnose children's SCPF by color Doppler echocardiography.

Objective To analyze the clinical characteristics of gravidas with HBV in Nanfang Hospital from 2008 to 2014. Methods 22 906 gravidas were retrospectively investigated. Results The HBsAg positive rates were 11.64% and 6.16% when the gravidas were divided into Cantonese and non-Cantonese groups (χ2 =193.370, P < 0.005). The ALT abnormal rates in HBeAg positive and HBeAg negative gravidas were 17.96% and 6.68% (χ2=62.594, P<0.005). Conclusion The HBsAg positive rate of gravidas in Guangdong and the ALT abnormal rate of HBeAg positive gravidas are higher.

Objective To establish the orthotopic bladder cancer model of multidrug resistance as the human' s, and detect its resistance condition. Methods Two groups of nude rats 4-6 weeks of age were inculated with 1×107 cell of T24 or T24-ADM, following with observation and putting down their meat, drink,mental condition, urine and abdominal mass growth. Animals were sacrificed after 4 weeks later, then their bladder were weighted and measured, histopathologic assessment was performed,mdr1 was detected by PCR,and cells from the bladder tumors were detected of multidrug resistence by MTT. Results Group of nude rats inculated with T24-ADM generated tumors about 80 % (8/10), the one inculated with T24 was 90 % (9/10)and about 2-3 days early. The blank group had no rats emerge tumors in bladder mucosa at all. Bladder weight and volume: (0.8±0.3) g, (1.0±0.5) g, (875±158) mm3, (903±192) mm3, difference between the two groups had no significant (t = 1.332 and t = 1.215, P＞0.05). Histopathologic detection: The two groups of bladder cancer tissue biopsies can be seen more chaotic arrangement of cell structure, cell body shape is irregular, to the depth of myometrial invasion in different without breaking the film. Between the two groups there were no significantly differences. PCR detection of mdr1 expression differences between the two groups was significant (t = 3.612, P ＜0.01). Cytological detection of drug-resistant cell volume is slightly larger, and no significant difference in morphology. MTT detection: cells from the inculated T24-ADM mice bladder tumor were more resistance to ADM than the ones from the inculated T24 mice bladder tumor (F = 412.107, P＜0.01), and for several other drugs were also resistant. Conclusion Cell transplantation was successfully used to establish bladder cancer model in situ of T24-ADM, and with multi-drug resistance characteristics. The model laid the foundation for further multi-drug resistance research of bladder cancer.

PURPOSE: The present study was designed to determine whether rapamycin could inhibit transforming growth factor beta1 (TGF-beta1)-induced fibrogenesis in primary lung fibroblasts, and whether the effect of inhibition would occur through the mammalian target of rapamycin (mTOR) and its downstream p70S6K pathway. MATERIALS AND METHODS: Primary normal human lung fibroblasts were obtained from histological normal lung tissue of 3 patients with primary spontaneous pneumothorax. Growth arrested, synchronized fibroblasts were treated with TGF-beta1 (10 ng/mL) and different concentrations of rapamycin (0.01, 0.1, 1, 10 ng/mL) for 24 h. We assessed m-TOR, p-mTOR, S6K1, p-S6K1 by Western blot analysis, detected type III collagen and fibronectin secreting by ELISA assay, and determined type III collagen and fibronectin mRNA levels by real-time PCR assay. RESULTS: Rapamycin significantly reduced TGF-beta1-induced type III collagen and fibronectin levels, as well as type III collagen and fibronectin mRNA levels. Furthermore, we also found that TGF-beta1-induced mTOR and p70S6K phosphorylation were significantly down-regulated by rapamycin. The mTOR/p70S6K pathway was activated through the TGF-beta1-mediated fibrogenic response in primary human lung fibroblasts. CONCLUSION: These results indicate that rapamycin effectively suppresses TGF-beta1-induced type III collagen and fibronectin levels in primary human lung fibroblasts partly through the mTOR/p70S6K pathway. Rapamycin has a potential value in the treatment of pulmonary fibrosis.
Cells, Cultured ; Collagen Type III/metabolism ; Fibroblasts/*drug effects/metabolism/physiology ; Fibronectins/metabolism ; Humans ; Lung/cytology/drug effects ; Pulmonary Fibrosis/drug therapy ; Signal Transduction/drug effects ; Sirolimus/*pharmacology ; TOR Serine-Threonine Kinases/metabolism/physiology ; Transforming Growth Factor beta1/*antagonists & inhibitors/physiology

Objective To evaluate the protective effect of pterostilbene against ultraviolet B (UVB)-induced acute damage in HaCaT cells,and to explore related mechanisms.Methods The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo1ium (MTS) assay and flow cytometry were performed to estimate the proliferative activity and the apoptosis and necrosis rate of HaCaT cells treated with different concentrations of pterostilbene respectively,so as to screen the non-toxic concentration of pterostilbene.HaCaT cells were randomly divided into several groups:normal control group receiving no treatment,UVB group irradiated with 57 mJ/cm2 UVB,3 pterostilbene groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours,3 pterostilbene + UVB groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours followed by UVB radiation.Western blot analysis was conducted to detect changes of the transcription factor NF-E2-related factor 2 (Nrf2) expression in cell nuclei and cytoplasm before and after the treatment with pterostilbene and UVB,quantitative PCR to determine the mRNA expression of catalase and superoxide dismutase in the HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to evaluate the activity of catalase and superoxide dismutase.Results MTS assay and flow cytometry showed that 2.44,4.88 and 9.75 μmol/L pterostilbene had non-toxic effect on HaCaT cells.The protein expression of Nrf2 in the nuclei and cytoplasm in the normal control group was 1.03 ± 0.08 and 1.04 ± 0.11 respectively.Compared with the normal control group,the protein expression of Nrf2 in the nuclei and cytoplasm experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups,and the UVB group showed similar protein expression of Nrf2 in the cytoplasm,but significantly increased protein expression of Nrf2 in the nuclei (1.77 ± 0.08,q =17.24,P ＜ 0.01).Compared with the normal control group and UVB group,the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups all showed significantly lower protein expression of Nrf2 in the cytoplasm (0.86 ± 0.10,0.87 ± 0.11 and 0.46 ± 0.11 respectively,all P ＜ 0.05),but significantly higher protein expression of Nrf2 in the nuclei (2.38 ± 0.11,2.57 ± 0.11 and 2.07 ± 0.13,all P ＜ 0.01).As qPCR showed,UVB radiation could significantly inhibit the mRNA expression of CAT (P ＜ 0.05),but had no obvious effect on the mRNA expression of SOD (P ＞ 0.05).The mRNA expression of CAT and SOD experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups compared with the normal control group (P ＞ 0.05).However,2.44,4.88 and 9.75 μmol/L pterostilbene could significantly reduce the inhibitory effect of UVB radiation on the mRNA expression of CAT (P ＜ 0.05) and up-regulate the mRNA expression of SOD in the pterostilbene + UVB groups (P ＜ 0.05).ELISA revealed that UVB radiation could inhibit the activity of CAT and SOD in the HaCaT cells (both P ＜ 0.001),while 2.44,4.88 and 9.75 μmol/L pterostilbene could reduce the inhibitory effect of UVB radiation on the activity of CAT and SOD (all P ＜ 0.05).However,the activity of CAT and SOD were still lower in the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups than in the normal control group (P ＜ 0.05).Conclusion Pterostilbene can prevent UVB-induced acute damage in HaCaT cells by activating the Nrf2 pathway and up-regulating the expression of the downstream antioxidant enzymes CAT and SOD.

Objective:To evaluate the effect of metformin on ultraviolet A (UVA) -induced photoaging of an immortalized human keratinocytes cell line (HaCaT), and to explore its potential mechanisms.Methods:Cell counting kit 8 (CCK8) assay was performed to evaluate the effect of metformin at different concentrations (0 - 100 mmol/L) on the viability of HaCaT cells, and 10 mmol/L metformin was selected for subsequent experiments. Cultured HaCaT cells were divided into a blank control group (conventional culture), a metformin group (treated with culture medium containing 10 mmol/L metformin), a UVA irradiation group (conventional culture for 24 hours followed by 10 J/cm 2 UVA irradiation) and a metformin + UVA group (treated with culture medium containing 10 mmol/L metformin for 24 hours followed by 10 J/cm 2 UVA irradiation) ; UVA irradiation was performed at a dose of 10 J/cm 2 once a day for 3 consecutive days. After 4-day treatment, cells were collected, the β-galactosidase assay was performed to determine the proportion of senescent cells in each group, 2′, 7′-dichlorodihydrofluorescein diacetate assay to detect levels of intracellular reactive oxygen species (ROS), and the comet assay to detect DNA damage levels. Additionally, some HaCaT cells were divided into the blank control group, metformin group, 1.25 μmol/L dorsomorphin (an adenosine monophosphate-activated protein kinase [AMPK] inhibitor) + metformin group, and 2.5 μmol/L dorsomorphin + metformin group, and cells in the latter two groups were treated with 1.25 and 2.5 μmol/L dorsomorphin respectively for 2 hours, followed by the treatment with 10 mmol/L metformin for 24 hours. Western blot analysis was performed to determine the cellular localization and phosphorylation levels of nuclear factor-erythroid 2-related factor 2 (Nrf2). By using the small-interfering RNA (siRNA) -mediated silencing method, siRNA-Nrf2 was transfected into HaCaT cells to knock down Nrf2 expression (siRNA-Nrf2 group) ; 2.5 μmol/L dorsomorphin-treated HaCaT cells or Nrf2-knockdown HaCaT cells were treated with metformin and UVA irradiation (dorsomorphin + metformin + UVA group, siRNA-Nrf2 + metformin + UVA group, respectively), and the proportions of senescent cells were further calculated in each group. Statistical analysis was carried out by using one-way analysis of variance and two-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:Treatment with different concentrations of metformin for 24 hours could affect the viability of HaCaT cells to varying degrees ( F = 5 206.31, P < 0.001) ; there were no significant differences in the relative survival rates of HaCaT cells between the 10 - 20 mmol/L metformin groups and the control group (0 mmol/L metformin group, all P > 0.05), while the relative cell survival rates were significantly lower in the 25 - 100 mmol/L metformin groups than in the control group (all P < 0.05). After UVA irradiation, HaCaT cells shrank significantly and became narrow and elongated, and the intercellular spaces increased; the relative cell survival rate was significantly lower in the UVA irradiation group (76.13% ± 1.03%) than in the blank control group (100.00% ± 1.24%, LSD- t = 14.86, P < 0.001), but significantly higher in the metformin + UVA group (106.69% ± 2.45%) than in the UVA irradiation group (LSD- t = 11.55, P < 0.001). Moreover, the UVA irradiation group showed significantly increased proportions of senescent cells (45.14% ± 4.98%), intracellular ROS levels (144.61% ± 4.91%), and percentages of DNA in the tail (75.33% ± 1.77%) compared with the blank control group (23.84% ± 1.89%, 55.49% ± 1.57%, 1.88% ± 0.29%, respectively, all P < 0.001), while the metformin + UVA group showed significantly decreased proportions of senescent cells (24.26% ± 1.34%), intracellular ROS levels (58.62% ± 2.17%), percentages of DNA in the tail (15.83% ± 1.23%) compared with the UVA irradiation group (all P < 0.001). Western blot analysis showed that the Nrf2 expression in the cytoplasm was lower in the 10 mmol/L metformin group than in the blank control group, while the phosphorylated Nrf2 expression in the nuclei was higher in the 10 mmol/L metformin group than in the blank control group, suggesting that metformin could effectively induce the phosphorylation of Nrf2 and its nuclear translocation; both the pretreatment with 1.25 and 2.5 μmol/L dorsomorphin could significantly reduce the phosphorylation levels of AMPKα and Nrf2 induced by 10 mmol/L metformin. The proportions of senescent cells in the dorsomorphin + metformin + UVA group and the siRNA-Nrf2 + metformin + UVA group were 67.84% ± 2.74% and 65.94% ± 1.33%, respectively, which were significantly higher than those in the metformin + UVA group (37.76% ± 1.64%, t = 14.45, 13.34, respectively, both P < 0.001) . Conclusion:Metformin may inhibit UVA-induced photoaging of HaCaT cells by activating the AMPK/Nrf2 signaling pathway, scavenging ROS and reducing DNA damage.

Objective:To investigate the value of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid- D-Phe1-Tyr3-Thr8-octreotide (DOTATATE) combined with 18F-FDG total-body PET/CT imaging in the diagnosis and heterogeneity assessment of primary foci of neuroendocrine neoplasms (NEN). Methods:Clinical, imaging and pathological data of 39 patients with pathological diagnosis (30 NEN and 9 non-NEN, 18 males and 21 females, age (54.0±11.4) years) who underwent 1/10 activity 18F-FDG and 1/2 activity 68Ga-DOTATATE total-body PET/CT combined imaging in Zhongshan Hospital, Fudan University from August 2020 to March 2023 were retrospectively analyzed. The NEN primary foci were classified as neuroendocrine tumor (NET) G1, G2, G3, and neuroendocrine carcinoma (NEC). Diagnostic efficacy of combined dual-low activity dual-tracer imaging for NEN primary foci and its value for evaluating tumor heterogeneity were analyzed. Results:The sensitivities, specificities, and accuracies of 68Ga-DOTATATE alone and in combination with 18F-FDG total-body PET/CT for the diagnosis of NEN primary foci were 81.2%(26/32), 7/9, 80.5%(33/41) and 90.6%(29/32), 7/9, 87.8%(36/41), respectively. Ten NET G1 and seven NET G2 lesions showed 68Ga-DOTATATE uptake and no 18F-FDG uptake; two NET G2 lesions showed no 68Ga-DOTATATE uptake but 18F-FDG uptake; and two NET G1 and six NET G2 lesions showed both 68Ga-DOTATATE and 18F-FDG uptake. The radiation doses of 68Ga-DOTATATE, 18F-FDG and a single examination of CT were (1.59±0.50), 0.49(0.44, 0.58) and 11.46(10.53, 12.85) mSv, respectively. Conclusion:Combining total-body PET/CT imaging with dual tracers can effectively diagnose NEN primary foci and assess inter-tumor heterogeneity.

Objective:To explore the feasibility of one-tenth dose 18F-FDG total-body PET/CT (TB PET/CT) in patients with malignant tumors. Methods:A retrospective analysis was carried out on 34 preliminarily diagnosed cancer patients (30 males, 4 females, age (64.0±1.6) years) who underwent one-tenth dose (0.37 MBq/kg) 18F-FDG TB PET/CT examination between April 2020 and September 2022 in Zhongshan Hospital, Fudan University. The raw data were reconstructed into 15 min and initial 2 min PET images (G15 and G2, respectively). A matched cohort of 34 preliminarily diagnosed malignant tumor patients (27 males, 7 females, age (63.3±2.1) years) undergoing full dose (3.70 MBq/kg) 18F-FDG conventional digital PET/CT (C PET/CT) examination with a PET scan rate of 2-3 min/bed position, were analyzed in line with the same pathological types. Signal-to-noise ratios (SNR) of G15, G2 and C PET/CT groups were compared, and based on the pathological results, the detection rates of those 3 groups for lesions were also compared. The χ2 test, independent sample t-test, Mann-Whitney U test, and Wilcoxon rank sum test were used for data analysis. Results:The significant differences in gender, age, body mass index (BMI), blood sugar level and postinjection waiting time between TB PET/CT group and C PET/CT group were not found ( χ2=0.98, t values: 0.08, -1.05, z values: 0.68, 0.41, all P>0.05). The SNR, from G15 to C PET and G2 groups, decreased gradually, which were 16.0(11.3, 20.0), 10.5(8.2, 13.5) and 8.4±0.3 respectively ( z values: 5.09, 3.31, -4.24, all P<0.05). All primary lesions and hepatic metastases were detected by G15 and G2 imaging (100%, 37/37) as well as by C PET/CT (100%, 36/36). The detection rates for lymph node metastasis lesions were 10/15 in the G2/G15 groups, which were higher than the detection rate in the C PET/CT group (64.4%(29/45); χ2=62.03, P=0.002). Conclusion:One-tenth dose 18F-FDG TB PET/CT with a 2-minute acquisition is feasibility in the clinical practice.