Objective To evalhate the protective effect of oral raloxifene on lung function after acute lung injury (ALI) in rats. Methods Thirty male adult Sprague-Dawley rats were used and divided into three groups: LPS raloxifene hydrochloric acid. group before secondary impact ( Group A, n = 10 ), LPS raloxifene hydrochloric acid group after secondary impact ( Group B, n = 10) and control group ( n = 10). All the rats were injected intraperitoneally with 5 mg/kg LPS. Raloxifene (30 mg/kg) was orally administered one hour before LPS injection and 14 hours after LPS injection in Groups A and B. The con-trol group remained free. All the animals were anesthetized by intraperitoneal injection of pentobarbital so-dium at 40 mg/kg and the femoral artery was cannulated 16 hours after LPS injection to measure the mean arterial pressure (MAP). All the rats received a direct intratracheal injection of hydrochloric acid ( pH = 1.2, 0.5 ml/kg). Before injection of hydrochloric acid and at 0. 5,1.5 and 4 hours after injection of hy-drochloric acid, the blood gas was measured. Fifteen rats ( five from each group) underwent a micro posi-tron emission tomography ( [18F] FDG microPET) scan of the thorax four hours after hydrochloric acid in-stillation. Then, the lung tissue was collected for histopathological examination. Results The Group B showed better pulmonary gas exchange and more stable MAP compared to the control group. The [18F] fluorodeoxyglueose uptake and histological lung injury score were 9. 01 ± 1.58 and 12.6 ± 0.97 respec-tively in Group B, which were higher than 4. 67 ± 1.33 and 9. 01 ± 1.58 respectively in control group (P < 0. 01 ). Conclusions Raloxifene exerts significant protective effect on lung function after ALI. [18F] FDG microPET is a useful method to evaluate the inflammatory reaction during ALI.

BACKGROUND:Lung cancers are highly heterogeneous and resistant to available therapeutic agents, with a five year survival rate of less than 15%. It has been difficult to determine the basis of lung cancer heterogeneity and drug resistance. Cancer stem cellmodel has attracted a significant amount of attention in recent years as a viable explanation for the heterogeneity, drug resistance, dormancy and recurrence and metastasis of various tumors. OBJECTIVE:To summarize the current understanding of lung cancer stem cells, including their histological types and tumor growth areas, and to discusses the prognosis of lung cancer and its relationship with lung cancer stem cells, in an effort to eradicate these cells to combat lung cancer. METHODS:In order to search relevant articles about the lung cancer stem celland its relationship with lung cancer from PubMed and Sciencedirect databases (from 1990 to 2014), a computer-based search was performed, using the key words of“lung cancer, cancer stem cell, lung cancer stem cell, lung cancer occur, tumor heterogeneity, drug resistance, gene mutation, signal pathways”in English. After eliminating literatures which were irrelevant to research purpose or containing a similar content, 48 articles were chosen for further analysis. RESULTS AND CONCLUSION:The cancer stem cellmodel has gained considerable support recently in context of lung cancers and stem-like cells that are associated with aggressive cancer behavior, metastatic progression, resistance to therapy and relapse. Since lung cancer stem cells are thought to consist of a heterogeneous population depending on the histology and site of tumors, and multiple signaling pathways might have to be targeted to effectively eliminate lung cancer stem cells for therapeutic benefit. It can be imagined that the multidisciplinary efforts currently under way to characterize and target stem-like cells in lung cancer wil reap significant therapeutic benefits in the future.

BACKGROUND:Studies have shown that lung cancer stem cel s can be isolated from lung cancer cel lines. But there are few reports about in vitro isolation, culture and identification of lung cancer stem cel s in patients with lung squamous carcinoma.
OBJECTIVE:To explore the feasible methods of harvesting lung cancer stem cel s from fresh lung cancer tissue in patients with lung squamous carcinoma.
METHODS:Side population cel s were isolated by col agenase digestion, Ficol density gradient centrifugation and Hoechst 33342 solution. The isolated cel s were suspended in conditioned medium for isolated culture. Flow cytometry method was used to detect lung cancer stem cel s based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+and CD133+/CD44+cel s were recorded.
RESULTS AND CONCLUSION:Cel s adhered at 0.5 hour after incubation;typical cel colony was formed at 4 days of culture;cel s showed paving stone-shape at 7 days in a total number of 10 8. The positive rates of CD133+, CD44+and CD133+/CD44+cel s at passage 4 were increased significantly. These findings indicate that stem cel-like lung cancer cel s were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which were stably and rapidly amplified in vitro, laying the foundation for the further study on the heterogeneity and resistance of lung cancer stem cel s in the future.

BACKGROUND:Studies have shown that lung cancer stem cels can be isolated from the lung cancer cel lines, But there are few reports on in vitro isolation, culture and identification of lung cancer stem cels in patients with lung squamous carcinoma.
OBJECTIVE:To establish the feasible methods of harvesting lung cancer stem cels from fresh lung cancer tissues in patients with lung squamous carcinoma, and to investigate the alterations in cel number and function during primary culture.
METHODS: Side population cels were isolated by colagenase digestion, Ficol density gradient centrifugation and Hoechst 33342 efflux properties. The isolated cels were isolated and cultured in conditioned medium. Flow cytometry method was used to detect lung cancer stem cels based on the cel surface markers CD133 and CD44, and the positive rates of CD133+, CD44+ and CD133+/CD44+ were recorded. The single cel clones assay, flat colony formation assay and the cel sphere formation assay were used to identify the stem-like characteristics of lung cancer stem cels between the first and fourth generations.
RESULTS AND CONCLUSION:The positive rates of CD133+, CD44+ and CD133+/CD44+ cels at the fourth generation were increased significantly, and the positive rates of CD133+ and CD133+/CD44+ cels at passage 4 were significantly higher than those at the first generation. The abilities of single cel clone formation, the flat colony formation and the cel sphere formation in the fourth-generation cels were greatly enhanced compared with the first-generation cels. Experimental findings showed that stem cel-like lung cancer cels were obtained from fresh lung cancer tissue in patients with lung squamous carcinoma, which stably and rapidly amplified in vitro, laying the foundation for the further study of the heterogeneity and drug resistance of lung cancer stem cels.

Objective To understand the epidemic status quo and influencing factors of Kashin-Beck disease among children in mountain areas of Jilin Province.Methods Two hundred eighty-two severe endemic areas in 18 counties were selected and stratified by random cluster sampling method,and the status quo of KashinBeck disease prevalence was investigated among 7-12 year-old children according to the Diagnostic Criteria of Kashin-Beck Disease (WS/T 207-2010).In the meantime,the annual household income and the proportion of economic crops replanted,grain out-sourced,and returning farmlands to forests and grass were surveyed in the disease affected areas.Results A total of 14 162 children were investigated who had no clinical symptoms.Among them,28 cases were detected positive using X-ray with a detection rate of 1.98‰,while most of the cases were metaphysis positive.The annual household income (≥5 000 Yuan vs.＜ 5 000 Yuan) in the year 2009-2011 had a significant impact on the incidence of Kashin-Beck disease (1.47‰ vs.3.67‰,x2 =6.179,P ＜ 0.05),while the areas of returning farmland to forests and grass which accounted ＞ 1％ had no significant influence on the incidence compared with that ≤ 1％ (3.30‰ vs.1.57‰,x2 =3.876,P ＞ 0.05);the areas of economic crops replanting which accounted ＞ 10％ had no significant influence on the incidence compared with that ≤ 10％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05);the proportion of grain out-sourcing which accounted ＞ 50％ had no significant influence on the incidence compared with that ≤50％ (3.07‰ vs.1.65‰,x2 =2.565,P ＞ 0.05).Conehision Up to 2012,the disease among 7-12 year-old children of the mountain areas of Jilin Province have basically met the standard of Kashin-Beck disease elimination and the situation remains stable;furthermore,the household income has a significant impact on the incidence of Kashin-Beck disease.

Objective To analyze the histological changes of bone diseases and to investigate the noninvasive measurements for diagnosing renal osteodystrophy (ROD) in maintenance dialysis patients . Methods Ninety-one patients were selected to receive bone biopsy . The bone samples were stained with HE, toluidine blue and Masson, and were examined with light microscopy . The levels of immunoreactive parathyroid hormone (iPTH), osteoprotegerin (OPG),sRANKL and osteocalcin (OCN) were determined in the patients enrolled from 2004 to 2006 . The level of iPTH was measured by radioimmunoassay . OPG and sRANKL were measured by ELISA,and OCN was measured by chemiluminescence . Results The incidence of ROD in the maintenance patients was 100% . According to the histological appearance, 50 cases (54 .9%) were high turnover bone disease (secondary hyperparathyroid bone disease), 9 cases (9 .9%) were low turnover bone diseases(osteomalacia and adynamic bone disease), and 32 cases(35 .2% ) were mixed bone disease . The level of iPTH in patients with ROD was significantly increased compared with healthy controls . It was the lowest in low turnover bone diseases . There was no difference among three types of ROD . OPG level was significantly increased compared with healthy controls [(2176 .58±1576 .08) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . The level in high turnover bone diseases was higher than that of the healthy controls [(2261 .85±1712 .22) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . There was no difference among three types of ROD .sRANKL level in high turnover bone disease was significantly increased compared with healthy controls [(0 .328±0 .524)pmol/L vs (0 .084±0 .190) pmol/L, P＜0 .05] . OCN level was also higher than that of the healthy controls (P＜0 .05), and the OCN level in low turnover ROD was the lowest among three types of ROD . OCN level in mixed ROD was dramatically increased as compared to low turnover ROD [(226 .633±66 .455) pmol/L vs (193 .03±104 .269) pmol/L, P ＜0 .05] .Conclusions The histological changes of bone disease can be indicated by iPTH level, but the types of ROD can not be distinguished according to iPTH level neither be differentiated by the levels of OPG, sRANKL and OCN . Bone histomorphometry is still the golden standard for diagnosing renal osteodystrophy .

Objective To identify the potential risk factors affecting mortality rate of ALl in severe trauma population. Method It was a retrospective cohort study treating trauma as a single cause for emergency depart-ment (ED)) and emergency intensive care unit (EICU) admissions. Eighteen potential risk factors affecting the mortality of ALI were examined by univariate and multivariate logistic analyses in these severe trauma patients. Re-sults There were 343 severe trauma patients with post-traumatic ALI admitted to ED and EICU the Second Affili-ated Hospital Medical College,Zhejiang University,during the study period. The five risk factors that affected the mortality with unadjusted odd ratios (ORs) and 95% confidence intervals (CIs) were (1) APACHE Ⅱ score, (2)duration of trauma, (3) age, (4) aspiration of gastric contents, and (5) DIC. Specific risk factors also affected different patients subpepulations at different degrees. Conclusions Factors of APACHE Ⅱ score and aspiration of gastric contents that can predict the mortality of ALl may exist in the early stage of trauma. Duration of trauma and DIC that greatly affect the short- and long-term development of ALI deserve special attention. Elderly patients (aged beyond 65 years) are the independent risk factor for the secondary sepsis and deterioration of pulmonary function. Patients with these risk factors need aggressive supportive care as early as possible in order to prevent fur-ther aggravation.

Objective To investigate the effect of isoflurane on the levels of protein kinase A (PKA) and protein kinase C (PKC) in hippocampus in rats. Methods Thirty-six 3-month-old male SD rats weighing 180-220 g were randomly divided into 3 groups ( n = 12 each): group Ⅰ underwent the cognitive function test without being pretreated with isoflurane inhalation (group C); group Ⅱ and Ⅲ inhaled 1.2% isoflurane for 4 h and underwent the cognitive function test 2 days and 2 weeks later respectively (group Ⅰso1,Iso2). Morris water maze was used to assess the cognitive function and the escape latency was recorded. The animals were killed immediately after the test.The hippocampus was isolated for determination of the expression and activities of PKA and PKC.Results The escape latency was significantly longer in group Ⅲ than in group Ⅰ.The expression of PKA and PKC was significantly down-regulated and the activities of PKA and PKC were significantly decreased in group Ⅱand Ⅲ as compared with group Ⅰ . There was no significant difference in the expression and activities of PKA and PKC between group Ⅱ and Ⅲ . Conclusion Four hour 1.2% isoflurane inhalation can decrease cognitive function by inhibiting the levels of PKA and PKC in hippocampus.

Objective To investigate the diagnostic and clinical significance of computerized tomography (CT) attenuation values (hounsfield unit,HU) in hydronephrosis with infection.Methods One hundred and eighty-five cases of upper urinary tract calculi with hydronephrosis from June 2014 to June 2016 were retrospectively analyzed.There were 82 males and 103 females with a mean age of (52.3 ± 13.1)years old,ranging 18-80 years old.58 cases suffered hydronephrosis without infection,55 cases suffered acute pyelonephritis and 72 cases suffered pyonephrosis.The CT attenuation values of the renal pelvis urine in three groups were measured.Results The CT attenuation value of hydronephrosis without infection group was (5.61 ± 3.67) HU,95 ％ CI(4.64-6.57) H U.In acute pyelonephritis group,CT attenuation value was (8.35 ± 5.63) HU,95％ CI(6.83-9.87) HU.In pyonephrosis group,the CT attenuation value was (13.92 ± 6.21) HU,95％ CI (12.46-5.38) HU.The CT attenuation value of pyelonephritis compared with that of hydronephrosis without infection was significant different.(P ＜ 0.01).The CT attenuation value of the patients with pyonephrosis was significantly higher than that of patients without infection and with pyelonephritis (P ＜ 0.01).Conclusions The CT attenuation value of renal pelvis urine can predict intrarenal infection.Furthermore,The measurement of CT attenuation value has some clinical significance in preoperative evaluation of hydronephrosis with infection.

Objective:To determine the expression of glucose transporter 3 (GLUT3) in cutaneous squamous cell carcinoma (cSCC), and to evaluate its effect on the cSCC cell line A431.Methods:From June 2016 to December 2020, 22 paraffin-embedded tissue specimens were collected from patients with pathologically confirmed cSCC in the Department of Dermatology, Peking University Third Hospital, and 20 discarded normal skin tissues after dermatological surgeries served as controls. Immunohistochemical assay was performed to determine the GLUT3 expression in cSCC tissues and normal skin tissues. Cultured A431 cells were divided into two groups: GLUT3 overexpression group transfected with a lentiviral vector carrying the SLC2A3 gene, and negative control group transfected with an empty lentiviral vector. Real-time fluorescence-based quantitative PCR and Western blot analysis were conducted to determine the mRNA and protein expression of GLUT3 in A431 cells in different groups, the cell proliferation assay (MTS assay) was performed to estimate the cell proliferative activity, and the live-cell analysis system Incucyte S3 was used for real-time evaluation of the migratory and invasive abilities of A431 cells in different groups. The relative glucose consumption and lactic acid production in A431 cells at 48 hours were measured by using glucose and lactic acid assay kits, retrospectively. Two independent samples t-test was used for comparisons between two groups, and one-way analysis of variance was used for comparisons among multiple groups. Results:The GLUT3 expression was significantly higher in the cSCC tissues than in the normal skin tissues (immunohistochemical assay score: 9.39 ± 2.56 points vs. 2.30 ± 2.60 points; t = 8.91, P < 0.05). Compared with the negative control group, the mRNA and protein expression of GLUT3 markedly increased in the GLUT3 overexpression group. MTS assay showed significantly increased proliferative activity of A431 cells in the GLUT3 overexpression group compared with the negative control group after 24- and 96-hour treatment ( t = 2.49, 3.54, P = 0.048, 0.012, respectively); cell fusion rates in the scratched area were significantly higher in the GLUT3 overexpression group than in the negative control group in the cell migration assay at 6, 12 18, and 24 hours and cell invasion assay at 12, 18, and 24 hours (all P < 0.05). At 48 hours, the relative glucose consumption and lactic acid production in A431 cells were significantly higher in the GLUT3 overexpression group than in the negative control group ( t = 2.98, 2.20, P = 0.011, 0.038, respectively) . Conclusion:GLUT3 was highly expressed in the cSCC tissues, and may participate in the occurrence and development of cSCC by providing energy to cSCC cells via promoting glucose uptake in cells to enhance their proliferative, migratory and invasive abilities.