AIM: To investigate the effects of phorbol ester (TPA) on the anti-tumor effect and renal toxicity of cisplatin (CP). METHODS: MTT assay was used to examine the effect of TPA on the proliferation inhibition of CP in A549 and SPC-A-1 lung cancer cells. Also the effect of TPA on acute toxicity of CP was observed by once injection of high dose CP through caudal vein; The tumor-bearing mice model was explored to investigate the effect of TPA on tumor inhibition ratio and renal toxicity of CP in vivo. And the effect of TPA on renal oxidative stress induced by CP was detected. RESULTS: 1 ng/mL TPA could significantly enhance the inhibitory effect of CP on cell proliferation. In acute toxicity test, TPA could significantly reduce the toxicity of CP and prolong the survival time of animals. And the tumor weight (P<0.05), serum creatinine (P<0.05) and urea nitrogen levels (P<0.01) in TPA combined with CP group were significantly lower than those in CP group. Meanwhile, the results of HE staining showed that the renal tissue damage was significantly reduced in the combined group compared with CP group. The contents of MDA in renal tissue were decreased (P<0.01). However, the contents of GSH and the activity of SOD were increased (P<0.05) in TPA and CP combined group. CONCLUSION: TPA can enhance the inhibitory effect of CP on cell proliferation and inhibit tumor growth in tumor-bearing mice. At the same time, TPA can reduce the renal toxicity of CP, which may be related to the inhibition of renal oxidative stress induced by CP.

Objective:To investigate the protective effect of lactoferrin(Lf) on lung injury in mice exposed to irradiation.Methods:C57BL/6 J mice were randomly divided into control group, 15 Gy irradiation group (IR group) and lactoferrin combined 15 Gy irradiation group (Lf+ IR group), with 5 mice in each group. The mice in the Lf+ 15 Gy group drank lactoferrin solution (10 mg/ml) from 3 days before irradiation and contained the whole experiments. Then, single chest 15 Gyirradiation was performed both in the IR and Lf+ IR groups. The body weight and other characteristics were monitored during the experiment. The mice were killed at day 14 after irradiation. The lung histopathology was observed by HE staining. Serum inflammatory cytokine such as HMGB1, TNF-α, IL-1β and IL-6 was determined by ELISA method . The expression of inflammatory related protein in lung tissue including HMGB1, TLR4, MyD88 and NF-κB were performed by immune histochemistry and Western blot method.Results:Compared with the control group, lung weight was significantly increased ( t=3.20, P<0.05), pulmonary hyperemia and inflammatory cell infiltration was observed in the IR group. Exposure also significantly increased serum level of TNF-α[(291.80±5.49) vs.(332.25±22.18)pg/ml]( t=3.07, P<0.05), up-regulated the expression of inflammatory related protein in lung tissue ( t=4.04, 4.78, 3.77, 6.14, P<0.05). Lactoferrin intervention (Lf+ IR group) significantly decreased lung weight ( t=2.18, P<0.05), alleviated histopathologic changes, decrease serum levels of HMGB1, TNF-α and IL-1β ( t=4.67, 2.97, 3.49, P<0.05). On the other hand, lactoferrin intervention decreased the positive cell number of HMGB1 and NF-κB, and down-regulated the protein expression of HMGB1, TLR4, MyD88 and NF-κB in lung tissues, with significant difference with the IR group ( t=8.06, 9.80, 3.07, 5.56, P<0.05). Conclusions:Lactoferrin plays the protective effect of radiation-induced lung injury through the downregulation of inflammatory response, such as HMGB1/TLR4/MyD88/NF-κB signaling pathway.

Objective: To introduce the method of galvanic vestibular stimulation-vestibular evoked myogenic potentials (GVS-VEMP) as well as to observe and analyze the parameters and elicited rate of GVS-cVEMP and GVS-oVEMP in healthy young people in China. Methods: Twenty six normal young subjects were recruited for conventional examinations of GVS-VEMP. The subjects were 21-37 years old, average age was (25.8±3.7) years old, including 13 males and 13 females. The galvanic stimulation intensity of 3 mA/1 ms was used to evoke cVEMP and oVEMP on the sternocleidomastoid and inferior extraocular muscles respectively, and the intensity of stimulus was decreased until the response disappeared, the threshold, latency, amplitude, interval phase and interaural amplitude ratio(IAR) were calculated. SPSS18.0 software was used for statistical analysis. Results: All subjects were elicited normal GVS-cVEMP and GVS-oVEMP under 3 mA/1 ms, the elicited rate was 100%. The threshold of GVS-cVEMP was (1.18±0.47) mA, p1 latency was (10.43±1.54) ms, n1 latency was (17.91±1.20) ms, the amplitude was (102.47±56.77) uV and IAR was (0.26±0.20). The threshold of GVS-oVEMP was (1.12±0.50) mA, n1 latency was (8.46±1.05) ms, p1 latency was (11.83±1.27) ms, the amplitude was (9.12±6.82) uV and IAR was (0.25±0.20). In terms of gender and lateral comparison, only the GVS-oVEMP amplitude was higher for male than for female, which had significant statistical difference (P<0.05), and there was no statistical difference in the other parameters between GVS-cVEMP and GVS-oVEMP. Conclusion GVS-cVEMP and GVS-oVEMP could be elicited in healthy youth population, and the parameters could provide reference for subsequent vestibular function evaluation.

Polycyclic aromatic hydrocarbons (PAHs) are widely found in the environment, and comparing to adults, children are more vulnerable to PAHs exposure. Urinary metabolites of PAHs are used as preferred biomarkers to estimate the PAHs exposure. Systematic review on the internal exposure level of children and adolescents is rare. We aimed to calculate the internal exposure levels of PAHs in children and adolescents and compare the levels of PAHs internal exposure in various children groups. We searched PubMed, OVID, Web of Science, EBSCO, ACS, and four Chinese databases, and all studies examining the urinary concentrations of PAHs in children and adolescent were identified. The total exposure level of 11 PAHs metabolites were pooled. Standard mean difference (SMD) and 95% confidence intervals (CIs) of PAHs urinary concentration were calculated and pooled by RevMan5.3 to compare the exposure levels of different children groups. We found that 1-OHPyr, 2-OHNap, 2-OHFlu, 3-OHPhe, and 4-OHPhe were five PAHs metabolites most commonly studied in existing studies in children, and their total exposure levels were 0.38 ± 0.98, 2.32 ± 4.83, 0.81 ± 1.54, 0.09 ± 0.14, 0.03 ± 0.10 μmol/mol creatinine, respectively. The meta-analysis showed that the levels of 1-OHPyr were higher in higher environmental exposure group (SMD = 0.21, 95% CI = 0.03~0.40), ETS exposure group (SMD = 0.31, 95% CI = 0.08~0.54), and 6~11 years group (SMD = 0.16, 95% CI = 0.09~0.24); the level of 2-OHNap (SMD = 0.27, 95% CI = 0.01~0.53) was higher in higher environmental exposure group; however, the levels of 3-OHPhe (SMD = - 0.34, 95% CI = - 0.57~- 0.12) and 4-OHPhe (SMD = - 0.48, 95% CI = - 0.69~- 0.28) were higher in lower environmental exposure group. The levels of 1-OHPyr (SMD = - 0.01, 95% CI = - 0.11~0.10) and 2-OHNap (SMD = 0.01, 95% CI = - 0.20~0.22) were not statistically different between boys and girls. In conclusions, we found that the internal diversity of PAHs existed in children and adolescents, and the level of 1-OHPyr in children and adolescents was in higher status compared with non-occupational people who do not smoke.

Objective: To test the mechanism and upstream pathway of outer hair cell apoptosis in Cadherin 23 (Cdh23) gene mutant mice. Method: The mutant Cdh23^erl/erl(erl) mice were collected as the study group, while the C57BL/6J (B6) mice were chosen as the control group. A total of 70 mice per group were used in this study. The study group and control group underwent auditory-evoked brainstem response (ABR) tests at the same age. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect outer hair cell(OHC) apoptosis. The qRT-PCR was conducted to test the expression of ER stress markers immunoglobulin-binding protein (BiP) and C/EBP homologous protein (CHOP) mRNA. The expression and location of BiP and CHOP protein in OHC were detected by immunostaining. The expression of BiP protein in cochleae was identified by Western blot. The expression and location of CDH23 protein in OHC were discovered by immunostaining. Results: The ABR thresholds in erl mice were significantly higher than those in B6 mice at the age of 1 and 3 months (both P<0.05). The surface preparation with TUNEL staining confirmed OHC apoptosis in erl mouse cochleae which showed a higher TUNEL positive cell ratio than B6 mouse(t=11.291, P<0.01). The ER stress marker Bip and Chop mRNA were upregulated in the erl mouse inner ear, when compared with those in the B6 mouse(both P<0.05). The BiP protein extracted from the erl mouse cochleae was significantly higher than that of B6 mouse measured by Western blot (t=3.66, P=0.02). Immunostaining showed that BiP and CHOP were highly detected in the OHC in erl mouse cochleae, and was mainly detected in the perinuclear region of OHC. However, a bare BiP and CHOP signal were shown in B6 mouse cochleae. The CDH23 protein was specifically localized at the top of the OHC in B6 mice, indicating the localization of the tip links in hair bundle stereocilia. On the contrary, the CDH23^erl protein was found to be localized from the top to the nuclei of the OHC in erl mice. Portions of the CDH23^erl proteins failed to reach the top of the hair bundles and remained in the OHC cytoplasm. Conclusion As the downstream response of the Cdh23 gene mutation, portions of the mutant CDH23^erl protein was accumulated in ER lumen resulting in the increase of ER loading and ultimately triggered ER stress and hair cell apoptosis in erl mouse cochleae.

Objective: To observe the parameters of the results of suppression head impulse paradigm (SHIMP) in healthy adults, and to provide reference for evaluating vestibular oculomotor reflex function in patients with peripheral vertigo. Methods: Fifty healthy adults, 22 males and 28 females, aged from 23-65 years, with an average age of (38.5±11.6) years, were recruited from January to March 2018. Parameters provided by the video head pulse software included the gains, the latency and the peak velocity of saccades, and comparison was made with head impulse paradigm (HIMP). Results: All subjects were elicited anti-compensatory saccades in SHIMP. The normal values of left and right gains were 1.02 and 1.10 in HIMP, and 0.93 and 1.01 in SHIMP respectively. The left and right saccades latency were (201.1± 50.8)ms and (187.0± 42.9)ms, and the peak saccadic velocity were (302.7±58.5)°/s and (291.5±46.5)°/s in SHIMP; there were small but significant difference between two sides about gains in HIMP and SHIMP, as well as latency in SHIMP(P<0.05); there were small but significant difference between HIMP and SHIMP about gains in ipsilateral(P<0.01); there were no significant difference between two sides about peak saccadic velocity in SHIMP(P>0.05). Conclusions SHIMP can be used for the examination of vestibular oculomotor reflex function in adult population. It is easy to be operated and is convenient for clinical application. Combined with head pulse test, the function of the semicircular canal can be evaluated together.

Objective: To estabilsh animal methods of bone-conducted vibration elicited cervical vestibular-evoked myogenic potentials (BCV-cVEMP) and ocular vestibular-evoked myogenic potentials (BCV-oVEMP) in healthy guinea pigs. Methods: Eleven healthy (250-350 g) and awake guinea pigs were selected and undertake conventional BCV-cVEMP and BCV-oVEMP examination in prone position. Parameters of waveforms were cauculated. Results: The BCV-cVEMP and BCV-oVEMP both could be elicited in 100% (22/22) in guinea pigs respectively, threshold was (85.5±10.8)dB SPL and (90.7±10.6)dB SPL for cVEMP and oVEMP; n1 latency was (4.5±1.3)ms and (4.3±1.5)ms for cVEMP and oVEMP; p1 latency was (5.8±1.4)ms and (5.6±1.7)ms respectively; n1-p1 interwave latency was (1.2±0.4)ms for cVEMP and (1.4±0.6)ms for oVEMP, amplitude was (21.5±17.3)μV and (24.0±16.3)μV respectively. Conclusion Both BCV-cVEMP and BCV-oVEMP can be successfully elicited in healthy guinea pigs.

Objective The inhibitor of differentiation 3 (Id3) is an important transcriptional regulation factor, which participates in tumorigenesis, cell proliferation, and cell apoptosis.β-catenin, as a central molecule of the Wnt signaling pathway, is critical for tumor development.This study aimed to evaluate the expressions of these two molecules and the regulatory effect of Id3 on β-catenin in different tumor cells.Methods Total RNA was extracted using the Trizol Reagent.The relative mRNA expression levels of Id3 and β-catenin in tumor cells were detected by quantitative real-timePCR(qRT-PCR).The recombinant eukaryotic expression vector pEGFP/Id3 with the human Id3 gene was transfected into A549, A549/ DDP and SW-480 cells using the non-liposome-mediated method.The protein expressions of Id3 and β-catenin were determined by Western blot.Results The expression of Id3 was significantly lower in the colorectal cancer cell lines SW-480 and HT-29 than in A549 and other tumor cells (P<0.05), but remarkably higher in nasopharyngeal carcinoma CNE and 5-8F cells than in other tumor cells (P<0.05).The expression of β-catenin was the highest in SW-480in comparison withother malignant tumor cells(P<0.05), and the second highest was in gastric cancer AGS and colorectal cancer HT-29 cell lines, but low in H446, A549, SPC-A-1, A549/DDP, and SK-MES-1 cell lines and extremely low or almost absent in CNE and 5-8F cells (P<0.05).After transfected with pEGFP/Id3, the cells showed a decreased volume, wrinkled membrane and absent refraction under the fluorescence microscope, which, however, were not observed in most of the cells transfected with the empty vector pEGFP.Compared with the control, the Id3/pEGFP group showed remarkably increased expressions of Id3 mRNA in the A549, A549/DDI, and SW-480 cells (1.24±0.12 vs 193.12±2.80, 1.09±0.11 vs 188.30±2.60, and 0.92±0.29 vs 19.08±0.59, P<0.01), and the expression of β-catenin was significantly down-regulated in the transfected SW-480 cells with an overexpression of Id3 (0.98±0.05 vs 0.32±0.03, P<0.01), but exhibited no statistically significant differences from those in the transfected A549 and A549/DDP cells (0.98±0.07 vs 1.04±0.08 and 0.98±0.05 vs 0.32±0.03, P>0.05).Western blot showed the same results.Conclusion The expression levels of Id3 and β-catenin vary in different tumor cell lines.Anabnormally high level of β-catenin is an important risk factor for colorectal cancer, and the down-regulatedexpression of β-catenin after eogenous transfection of Id3 may provide some new ideas for target gene therapies of colorectal cancer.

Objective Rheumatoid arthritis (RA) is a typical type Ⅲ hypersensitivity with a large number of immune complexes (IC) and complement deposits in the synovial tissue , but its specific pathogenesis is not yet clear.This article was to explore the expression of the antigenic profile of serum ICs in RA.Methods ICs were isolated from the serum of 55 patients with RA (41 cases of anti-CCP antibody [+] and 14 cases of anti-CCP antibody [-]), 41 with systemic lupus erythematosus (SLE), and another 41 healthy controls by polyethylene glycol (PEG) precipitation, separated by immunoprecipitation, digested with trypsin in gel, and then subjected to mass spectrometry for identification.The levels of total proteins were compared among different groups using Vennny 2.1.0.The protein expression was considered to be up-regulated when the total protein level of the RA group was >2 times and down-regulated when it was <0.5 times that of the control.Further functional analysis was performed on the differential proteins in RA using the STRING software.Results Totally, 277 proteins were identified in the serum ICs of the RA patients, including 162 in the anti-CCP (+) and 248 in the anti-CCP (-) RA group.Compared with the SLE and healthy control groups, only 129 proteins were found in the RA patients, including 38 in the anti-CCP (+), 109 in the anti-CCP (-) RA group, and 18 in both the two groups.Among the proteins identified in the RA patients and healthy controls, 2 and 11 were up-regulated while 17 and 21 down-regulated in the anti-CCP (+) and anti-CCP (-) RA group, respectively.Conclusion More differentially expressed proteins were identified in the anti-CCP (-) than in the anti-CCP (+) RA patients.The identification of differentially expressed proteins provides a new idea and direction for the investigation of the pathogenesis and new biomarkers of RA.

Objective To scan protein expression profile of immune complexes (ICs) derived from the synovial tissue of the patients with rheumatoid arthritis (RA) based on liquid chromatography-tandem mass spectrometry (LC-MS).Methods The samples of synovial fluid were obtained from knee joints of the patients with RA and osteoarthritis (OA) used as control during therapeutic arthrocentesis in knee jiont at the Department of Orthopedics of Jinling Hospital,School of Medicine,Nanjing University.The protein expression profile of ICs was identified by enrichment strategy based on immunoprecipitation and LC-MS analysis.The value of fraction of total (FOT) was used to estimate protein abundance and screen the up-and down-regulated proteins.The function enrichment,interaction network and signal pathway of differential proteins were analyzed using softwares David and String.Results A total of 511 and 526 protein spots in ICs of RA and OA patients were identified respectively.Among them,170 proteins existed only in RA group.45 and 85 proteins in RA group were statistically up-and down-expressed compared with controls.Conclusion HSP90AA1,HSP70,HLAG,Thioredoxin,Annexin A2 and vitronectin may be involved in the pathogenesis of RA through different paths and possible to become promising diagnostic indicators or new therapeutic targets for RA.