Objective To elucidate the relationship between f luoride and cell apoptosis of ameloblasts and to further study the mechanism of dental fluorosis. Methods By establishing an experimental m odel of fluoride intoxication in pregnant C57BL/6N, we observed apoptosis in the fetal secretory ameloblasts in mice by means of transmission electron microscop e and TUNEL. Results High fluoride exposure in pregnant mic e could cause subameloblastic cysts in first lower molar of fetal mice. Programm ed cell death (PCD) occurred in the disturbed ameloblasts above cysts. Conclusion Intoxicat ion mechanism of fluoride on ameloblasts may occur by means of PCD.

Objective To determine the levels of total antioxidative capacity(TAOC) and malondialdehyde(MDA) in liver exposed to hyperoxia,and to explore whether oxygen inhalation could cause liver injury in newborn rats.Methods Sixty-four newborn rats which were less than 12-hour-old were enrolled in this study.The rats were randomly divided into hyperoxia group(FiO2=0.85,n=32) and control group(air,n=32).Eight rats in each group were randomly sacrificed to obtain liver tissues at 1d,3d,7d and 14d.The TAOC of liver homogenates was detected by chemical colorimetry,and the MDA level of liver homogenates was measured by thiobarbituric acid test.Results In the hyperoxia group,TAOC in liver increased on the 1st day[(3.60±0.28)U/mg prot vs(3.39±0.19)U/mg prot,P<0.05];TAOC began to decreased on the 3rd day,and significantly lower than that of control group on the 14th day [(3.10±0.15)U/mg prot vs (3.56±0.14)U/mg prot,P<0.01].In the hyperoxia group,the MDA level increased on the 3rd day[(3.58±0.11)nmol/mg prot vs(2.82±0.14)nmol/mg prot,P<0.01],and reached a peak on the 7th day[(3.58 ±0.11)nmol/mg prot vs(2.82±0.14)nmol/mg prot,P<0.01],then decreased but still remained higher than control group on the 14th day [(2.92±0.18)nmol/mg prot vs(2.77 ±0.09)nmol/mg prot,P<0.01].Conclusion Too more MDA in liver and TAOC decrease may cause liver injury in newborn rats exposed to hyperoxia.With the oxygen inhalation time prolonging,the liver injury aggravation.

Carcinoma ; pathology ; Carcinoma, Papillary ; Histiocytosis, Langerhans-Cell ; pathology ; Humans ; Thyroid Neoplasms ; pathology

Objective To investigate the effect of early low-glucocorticoid on hemodynamics and prognosis in the patients with septic shock.Methods Sixty patients with septic shock failing in active fluid resuscitation and vasoactive drugs in our hospital from June 2013 to August 2015 were selected and divided into the control group,early-hormone group and late-hormone group.MAP,HR,PO2/FIO2 and serum lactic acid levels were monitored in all selected patients before treatment and at 12,24,48 h after treatment.Apache Ⅱ,SOFA scores were assessed before treatment and on 1,3,7 d after treatment.The ventilation time,ICU stay time,hospital stay time and intravenous use time of vasoactive agents(VDNT) were recorded.Results The Apache Ⅱ scores and SOFA scores on 3,7 d after treatment in the early-hormone group were significantly decreased compared with the late-hormone group and control group (P＜0.05).MAP and HR at 24,48 h after treatment in the early-hormone group were significantly improved compared with the late-hormone group and control group (P＜0.05).The level of serum lactic acid at 12,24 h after treatment in the early-hormone group and late-hormone group were obviously lower than that in the control group,the levels of serum lactic acid at 12,24 h after treatment in the early-hormone group were obviously lower than those in the late-hormone group (P＜ 0.05).PO2/FIO2 at 12 h after treatment in the early-hormone group and late-hormone group were obviously better than that in the control group,and PO2/FIO2 at 12 h after treatment in the early-hormone group was obviously better than that in the late-hormone group(P＜0.05).The ventilation time,ICU stay time,hospital stay time and VDUT in the early-hormone group were significantly shortened compared with the late-hormone group and control group.The ventilation times,ICU stay time and VDUT in the latehormone group were significantly shortened compared with the control group (P＜0.05).Conclusion Early using low-dose glucocorticoid may restore hemodynamics more quickly,protects the organ function and improves the prognosis in the patients with septic shock.

Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.

Objective To explore the subcellular localization of Galectin-9 and its effect on allogeneic immune response.Methods The plasmid pEGFP-N1 was inserted with Galectin-9 fragment which was amplified from pBKCMV-Galectin-9 by PCR.The recombinant plagmid wag then transfected into CHO cells using JetPEI in vitro.The cells were cultured in G418 selecting mediam to obtain the stably-transfected cells.The transcription and expression of Galectin-9 gene were verified by immunohistochemical staining and RT-PCR.The solid-phase transgenic CHO cells or freshly-cultured supernatant wag added into the mixed lymphocyte response system to detect the inhibitory effect of Galectin-9.Galectin-9 protein wag administered intraperitoneally for 7d consecutively.Results The expression of Galectin-9 wag localized in the cytosol of CHO.The allogeneic mix lymphocyte proliferation was dose-dependently inhibited by the freshly-cultured supernatant from stably-transfected CHO cells.Furthermore,the supernatant from stably-transfected CHO cells dose-dependently inhibited the level of IL-2.The inhibitory effect could be reversed by Tim-3-Fc blocking.Administration of Galectin-9 significantly prolonged the survival of allogeneic cardiac transplants[(22.7±1.2)d vs(7.2±0.4)d)].Conclusion Galectin-9 may be secreted in physical situation to exert its immunomodulatory function on allogeneic immune response.Furthermore.Galectin-9 may be a novel therapeutic drug in transplant medicine.

Objective To study the application value of modified urine nucleoside's detection in prognosis of patients with bladder cancer. Methods We enrolled 85 patients with bladder transitional cell carcinoma confirmed by pathological examination.The 85 patients fulfilled one-year follow-up visit after TUR-BT and were reviewed every three months.The 85 patients did not relapse in the first third month after operation.At the sixth month after operation,20 cases relapsed.18 cases and 19 cases relapsed at the ninth month and the twelfth month after operation.Patients with recurrence added up to 57 cases as the recurrent group.The remaining 28 cases did not relapse at one year after operation as the no recurrent group.Of the 85 cases,55 cases were in T(is) - T1,while 30 cases were in T2 - T4.Of the 85 cases,27 cases were with G1,40 cases were with G2 and 18 cases were with G3.In T(is) -T1,there were 35 cases in recurrent group,while there were 20 cases in the no recurrent group.In T2 -T4,there were 22 cases in recurrent group,while there were 8 cases in the no recurrent group.There were 50 normal people in the control group.Highperformance liquid chromatography/electrospray ionization-quadrupole-time-of-flight mass spectromerry was used to measure the levels of change of two urine modified nucleosides (M1A,1-MeI) which the patients with bladder cancer had different pathology grades,clinical stages,before or after operation and recurrence or no recurrence. Results The levels at third month after operation in no recurrent group ( M1A:3.24 ± 0.40,1 -MeI:5.73 ± 0.67 ) were significantly lower than that before operation ( M 1A:4.34 ± 0.98,1-MeI:14.22 ± 4.05,P ＜ 0.005 ),and remained in low status at another time points after operation.The levels at the third month after operation in recurrent group (M1A:3.31 ±0.33,1-MeI:5.67 ±0.55) were significantly lower than that before operation ( M1A:4.32 ± 1.19,1-MeI:14.31 ± 4.12,P ＜ 0.005 ),which was on the rise and indicating a high level approaching the condition before operation.According to the time point before the operation,recurrent group and no recurrent group were higher than control group (M1A:2.91 ±0.84,1-MeI:5.56 ± 1.25,P ＜ 0.01 ).The levels at the sixth month,ninth month and twelfth month after operation in recurrent group ( M 1A referring to 4.04 ± 0.48,4.11 ± 0.47,4.09 ± 0.53 ;1-MeI referring to1 1.46 ± 1.34,12.14 ± 1.22,12.33 ± 1.27) were the highest (P ＜ 0.01 ).The levels of change of two urine modified nucleosides between pathology grade and clinical stage had no statistical difference ( P ＞ 0.01 ).The levels in recurrence group in T(is) - T1 ( M1 A:5.92 ± 1.28,1-MeI:20.01 ± 8.53 )were higher than the levels in no recurrent group ( M1A:4.02 ±1.22,1 -MeI:11.21 ± 6.45,P ＜ 0.05 ),which was the same in T2 - T4. Conclusion Urine modified nucleosides detection offer a certain clinical value the prognostic of operated bladder cancer patients.

Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P ＜ 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P ＜ 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P ＜ 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.

Objective To explore the high-risk factors of metabolic bone disease (MBD) in premature infants by retrospective analysis of the clinical data so as to provide evidence for optimal clinical management. Methods Clinical data of premature infants with birth weight<1500 g admitted in our hospital from January 2016 to December 2017 were retrospectively analyzed. Infants with serum alkaline phosphatase ( ALP )>500 IU/L and blood phosphorus <1. 5 mmol/L were selected as MBD group and premature infants with birth weight <1500 g were selected randomly as non-MBD group. General data, pulmonary surfactant, continuous positive airway pressure, mechanical ventilation, start time of enteral nutrition, parenteral nutrition ( PN) time, breast feeding time and breast milk fortifier adding, drug usage, hospitalization time and complications were re-corded and compared between the two groups. Results A total of 440 premature infants with birth weight<1500 g were admitted to the hospital during the study period. 58 [ 13. 2％ ( 58/440) ] infants were enrolled in the MBD group, among which infants with birth weight<1000 g accounting for 56. 9％ ( 33/58) . High birth weight (OR=0. 62, 95％ CI:0. 389-0. 990) was an independent protective factor of MBD in premature in-fants. The higher the birth weight, the lower the risk of MBD in premature infants. The longer duration of breast feeding time ( OR= 2. 191, 95％ CI:1. 628-2. 950) , later initial time of enteral feeding ( OR=2. 695, 95％CI:1. 710-4. 248), longer duration of PN (OR=6. 205, 95％ CI:3. 359-11. 463) time, longer duration of respiratory supporting time ( OR=1. 046, 95％ CI:1. 026-. 067) , longer hospital stay time ( OR=1. 703, 95％ CI:1. 109-2. 615) and small for gestational age ( OR=2. 965, 95％ CI:1. 163-5. 658) were inde-pendent risk factors of MBD in premature infants. The duration of PN was the most important independent risk factor of MBD in premature infants (OR=6.205, 95％ CI: 3.359-11.463). Conclusion Multiple factors can lead to MBD of premature infants. The high birth weight is an independent protective factor of MBD and the duration of PN is the most important independent risk factor of MBD in premature infants.

Objective:To investigate the value of bronchoalveolar lavage fluid (BALF) genechip analysis for the identification of pathogens in children with refractory pneumonia.Methods:A retrospective study of 500 children clinically diagnosed with refractory pneumonia in the Department of Respiratory and Critical Care Medicine, Kunming Children′s Hospital, Kunming Medical University between January 2020 to January 2022 was made.During hospitalization, bronchoscopic examination and bronchoalveolar lavage were performed.BALF was collected and analyzed using genechip technology to detect potential pathogens.At the same time, bacterial culture tests of sputum and BALF samples from the patients were performed. χ2 test was used to compare the positive rates of pathogens detected by different detection methods. Results:Of the 500 children patients, 482 cases (96.4%) were positive of BALF genechip analysis for pathogen identification.There were 71 cases (14.7%) infected with a single pathogen, and 411 cases (85.3%) with 2 or more pathogens.The top 3 bacteria were Streptococcus pneumoniae [117 cases (8.3%)], Haemophilus influenzae [63 cases (4.5%)], and Bordetella pertussis [32 cases (2.3%)]. The patients were mostly infected with respiratory syncytial virus [269 cases (19.1%)], followed by parainfluenza virus [217 cases (15.4%)], and adenovirus [132 cases (9.3%)]. Among the 500 patients, 116 cases (23.2%) were positive of BALF genechip analysis for bacteria identification, 47 cases (9.4%) had a positive BALF culture, 43 cases (8.6%) had a positive sputum culture.The bacterial detection rate of BALF genechip analysis was statistically significantly higher than that of BALF culture and sputum culture tests ( χ2=34.90, 39.85; all P<0.001). Conclusions:Most patients with refractory pneumonia have mixed infections.The genechip technology can rapidly and efficiently identify the pathogens, thus providing clinical guidance for anti-infection treatment.