ObjectiveTo observe the interaction of the calcifying nanoparticles (CNP) with human renal tubular epithelial cells (HK-2) in vitro,to observe and investigate the mechanisms of HK-2 injury induced by CNP,and to explore the potential role of CNP in the formation of Calcium oxalate kidney stones.MethodsHuman renal tubular epithelial cells were cultured in vitro and CNP was then added to the culture medium,the cell-crystal reaction was detected by light microscopy and transmission electron microscopy (TEM).To investigate the oxidative stress,NADPH oxidase inhibitor apocynin was chosen as the intervener.The levels of LDH,MDA,HA in the mediums after 24 h were assessed. ResultsCNP could induce changes of the HK-2.Adhesion and phagocytosis of CNP by the HK-2 were observed under TEM.After 24 h,the levels of LDH,MDA,HA were significantly different among the 4 groups ( P ＜ 0.05 ). ConclusionsHK-2 has abilities of adhering and phagocyting with CNP.CNP can cause damage induced by oxidative stress of HK-2.

Objective To examine the expression of heparin binding-epidermal growth factor-like growth factor (HB-EGF) in paclitaxel-resistant ovarian cancer and elucidate the relationship between HB-EGF and the resistance of ovarian cancer to paclitaxel. Methods The human ovarian carcinoma cell line A2780 and the paclitaxel-resistant human ovarian carcinoma cell line A2780/Taxol were cultured in vitro. Western blot was used to dectect the expression of HB-EGF protein in A2780 and A2780/Taxol groups. The A2780 cells were treated with cross-reacting material 197 (CRM197 and A2780 + CRM197 group) or dimethyl sulphoxide (DMSO;A2780 group), while the A2780/Taxol cells were treated with CRM197 (A2780/Taxol+CRM197 group) or DMSO (A2780/Taxol group). The effects of CRM197 on growth and proliferation was tested by methyl thiazolyl tetrazolium( MTT) and the results were showed as absorbance (A).The effects of CRM197 on cell cycles was tested by flow cytometry, while the effects of CRM197 on apoptosis was examined by caspase-3 activity assay and the results were showed as p-nitroaniline(pNa). In animal experiment, four groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability following CRM197 treatment. Immunohistochemistry was used to determine the expression of HB-EGF protein in A2780 and A2780/Taxol group. Results The expression level of HB-EGF protein in A2780/Taxol group (2.11±0.41) was significantly higher than that of A2780 group (0.75±0.20;P<0.01). The inhibition effect of CRM197 on the cell growth of A2780+CRM197 and A2780/Taxol+CRM197 group was accompanied by the acceleration of CRM197 concentration(P<0.01). When CRM197≥1 μg/ml, the inhibition effect of CRM197 on the cell growth of A2780/Taxol+CRM197 group was significantly higher than that in A2780/Taxol group(P<0.05). In cell cycle experiment, CRM197 induced the cell-cycle arrest at the G0/G1 phase in A2780+CRM197 cells[(67 ± 4)%] compared with A2780 cells[(54 ± 6)%;P<0.01], while CRM197 significantly induced the cell-cycle arrest at the G0/G1 phase in A2780/Taxol+CRM197 cells [(72± 4)%] compared with A2780/Taxol cells [(24±8)%;P<0.01]. CRM197 treatment in A2780+CRM197 group [(40 ± 6)μmol/L] led to the acceleration of the caspase-3 activity when compared to A2780 group [(6 ± 6)μmol/L;P<0.01], while CRM197 treatment in A2780/Taxol+CRM197 group [(66 ± 12)μmol/L] led to significant acceleration of the caspase-3 activity when compared to A2780 group [(9 ± 6)μmol/L;P<0.01]. In experiments in vivo, the expression scores of HB-EGF protein in A2780/Taxol tumors(10.8 ± 3.3) were higher than that in A2780 tumors (5.0±2.2;P<0.01). The tumor size and tumor weight of the A2780/Taxol+CRM197 group were both higher than those of the A2780+CRM197 group [(546 ± 85) mm3 vs (1 355 ± 119) mm3,(0.56 ± 0.09) g vs (1.31 ± 0.27) g; all P<0.01]. The CRM197 inhibition rate of the A2780+CRM197 and A2780/Taxol + CRM197 group were 43% and 68% respectively, showed that CRM197 significantly suppressed the growth of A2780/Taxol xenografts in vivo(P<0.01). Conclusions HB-EGF is over-expressed in paclitaxel-resistant ovarian cancer and may be contributes to drug resistance. Inhibition of HB-EGF expression potently enhances apoptosis and inhibit the growth of paclitaxel-resistant ovarian cancer, shedding light on the HB-EGF-targeted therapy options for chemoresistant ovarian cancer patients.

With literature of professor CHEN Yinglong such as Medical Notes of Doctor CHEN Yinglongfor Taiwan Compatriots,the'experience of doctor CHEN Yinglong treating common diseases in Fujian and Tai-wan is summarized. The diseases in the paper are constipation, retention of urine, asthma, vec6rdia, bi syndromeinsomnia, thoracic obstruction, pediatric obesity and freckle of face, etc. It is discovered that professor CHENYinglong treated diseases with combination of acupuncture and medication and accurate acupoints according to thefeature of climate in Fujian and Taiwan.
Acupuncture Points ; Acupuncture Therapy ; history ; Asthma ; therapy ; Constipation ; therapy ; History, 20th Century ; Humans ; Taiwan

Objective To investigate the effect and mechanism of CRM197, the heparin binding-epidermal growth factor-like growth factor (HB-EGF) inhibitor, on the reverse of the resistance ofovarian cancer to paclitaxel. Methods (1)The effect of CRM197 on the 50% inhibitory concentrations (IC50) of human ovarian carcinoma cell line A2780 and paclitaxel-resistant ovarian carcinoma cell line A2780/Taxol was tested by methyl thiazolyl tetrazolium (MTT) assay. Western blot was used to detect the effect of CRM197 on the expression of HB-EGF,epidermal growth factor receptor (EGFR) and plasma membrane glycoprotein (P-gp) protein in A2780 and A2780/Taxol cells. Real-time PCR was used to examine the MDR1 mRNA expression in these cells. (2) A2780/Taxol cells were divided into 4 groups, including the cells transfected with empty vector and saline treatment (empty vector group), MDR1 small interference RNA (siRNA)vector and saline treatment (MDR1 siRNA group),empty vector and CRM197 treatment (empty vector+CRM197 group) and MDR1 siRNA vector and CRM197 treatment (MDR1 siRNA+CRM197 group), respectively. Flow cytometry was used to detecte the effect of intracellular rhodomine 123 (Rh123) accumulation, and caspase-3 activity assay was used to test the effect of apoptosis in four groups of A2780/Taxol cells. (3) In experiments in vivo, A2780/Taxol cells were inoculated to nude mouse subcutaneously to determine the EGFR and P-gp protein expression following CRM197 treatment by immunohistochemistry. Results (1)In vitro,MTT examination showed that the IC50 of A2780/Taxol cells to paclitaxel in A2780/Taxol+CRM197 group [(6.4±0.3)μmol/L] was significantly lower than the IC50 in A2780/Taxol group [(34.1± 0.5)μmol/L,P<0.01], and the reveral fold of CRM197 was 5.3. The expression level of HB-EGF protein in A2780/Taxol+CRM197 group (1.44 ± 0.29) was significantly lower than HB-EGF protein in A2780/Taxol group (2.72 ± 0.32),respectively (P<0.05). The expression level of EGFR protein (0.71 ± 0.25) and P-gp protein (0.82±0.19) in A2780/Taxol+CRM197 group was significantly lower than EGFR protein (1.87±0.31) and P-gp protein (1.84 ± 0.27) of A2780/Taxol group (P<0.05). Compared with A2780/Taxol group(1.78 ± 0.27), MDR1 mRNA was significantly down-regulated in A2780/Taxol+CRM197 group (0.79 ± 0.13,P<0.05). (2) The fluorescence intensity of Rh123 of the A2780/Taxol cells in empty vector group, MDR1 siRNA group,empty vector+CRM197 group, MDR1 siRNA+CRM197 group was 33.4±1.6, 56.3±3.3, 43.5± 3.1,100.4 ± 7.4, and the pNA of the A2780/Taxol cells was(11.4 ± 1.2),(52.8 ± 0.9),(71.2 ± 3.6),(82.7 ± 3.8)μmol/L. The expression levels in MDR1 siRNA+CRM197 group were both higher than the expression levels in empty vector+CRM197 group, and the expression levels in empty vector+CRM197 group, MDR1 siRNA group were both higher than the expression levels in empty vector group (P<0.05). (3) In vivo, the expression scores of EGFR protein in A2780/Taxol+CRM197 tumors (4.4 ± 1.4) were lower than that in A2780/Taxol tumors (10.2 ± 3.1,P<0.05). The expression scores of P-gp protein in A2780/Taxol+CRM197 tumors (3.8 ± 1.1) were lower than that in A2780/Taxol tumors (8.8 ± 2.7,P<0.05). Conclusion CRM197 reverses the resistance of ovarian cancer to paclitaxel by increasing caspase-3 activity to advance apoptosis via EGFR/MDR1/P-gp pathway.

Objective To investigate the anti-inflammatory and analgesic effects of toothache drop pills. Methods The acute inflammatory models such as xylene-induced ear edema and egg white-induced paw edema and the chronic inflammatory model granuloma induced by cotton pellet implantation were used in researching the inflammatory effects of toothache drop pills. Meanwhile, the analgesic effects of toothache drop pills were observed by hot plate and acetic acid writhing test. Results Compared with the blank control group,the degree of ear swelling in mice in the positive control group, toothache drop pills middle and high dose group (2.56 ± 1.35 mg, 4.26 ± 1.21 mg, 3.23 ± 1.25 mg vs. 8.25 ± 1.21 mg) were lower than the blank control group (P<0.05). 120 minutes after administration,compared with the blank control group, the degree of paw swelling in mice in the positive control group, toothache drop pills middle and high dose group (23.54 ± 9.12 mg, 27.58 ± 9.14 mg, 21.25 ± 8.45 mg vs.39.54 ± 8.89 mg) were lower than the blank control group (P<0.05). Granuloma swelling quality in mice caused by cotton in the positive control group, toothache drop pills middle and high dose group (6.51 ± 2.58 mg, 7.82 ± 1.57 mg, 6.58 ± 3.47 mg vs. 13.58 ± 3.25 mg) were lower than the blank control group (P<0.05). The threshold of pain in mice in the positive control group, toothache drop pills middle and high dose group (20.86 ± 2.58 s, 20.25 ± 2.14 s, 20.75 ± 1.78 s vs.17.21 ± 3.31 s) were increased (P<0.05). The number of mice writhing in the positive control group, toothache drop pills middle and high dose group (23.47 ± 7.57, 28.65 ± 6.54, 24.36 ± 7.78 vs. 40.96 ± 6.58) were decreased(P<0.05). Conclusion Toothache drop pills had obvious anti-inflammatory and analgesic effects.

Objective To prepare toothache drop pills and establish its quality standard.Methods The drop pills were prepared by a routine method; HPLC was used for the determination of the EU isoimperatorin content.Results The drop pills were well- distributed in size, smooth and glossy in appearance, mild in hardness; TLC can identify isoimperatorin characteristic spots; Determination of the Indigo 0.50~520μg/ml linear relationship was good,r=0.999 8, the recovery of 98.38%.RSD values were 1.05%.Conclusion The preparation method of toothache drop pills is simple, and the drop pills are well-shaped with controllable quality.

Objective:To investigate the efficacy and the clinical value of magnetic resonance elastography(MRE) in diagnosis of hepatic fibrosis with Meta-analysis, and to provide basis for clinical treatment of hepatic fibrosis.Methods:The studies published before February 2, 2017 about MRE and staging of hepatic fibrosis in Chinese or English were retrived in the databases including PubMed, EMBase, Web of Science, Cochrane Library,CNKI, CBMDisc,VIP, Wanfang data, and supplemented by manual retrieval for relevant literatures.The inclusion and exclusion criterions were used to select and extract the literatures.The literatures qualitie were valuated based on QUADAS-2 tool.The sensitivity(SEN), specificity (SPE), diagnostic odds ratio (DOR), positive likelihood ratio (+LR), negative likelihood ratio (-LR) on the groups of F0 vs F1-F4,F0-F1 vs F2-F4,F0-F2 vs F3-F4, F0-F3 vs F4 and heterogeneity were combined and tested with Stata software respectively.HSROC and AUROC were also implemented.Results:A total of 1 332 studies were searched, and 22 were included.21 of them were in English and 1 in Chinese.The results of Meta analysis showed that the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0 vs F1-F4 group were 88.8%(85.0-91.7),95.9%(91.5-98.0),21.435(10.215-44.979),0.117(0.086-0.159),183.187(72.533-462.650) and 0.96(0.94-0.98) ,respectively;the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0-F1 vs F2-F4 group were 93.3%(89.2%-35.9%), 94.1%(90.2%-96.5%),15.839(9.344-26.848),0.072(0.044-0.117),221.224(100.980-484.648) and 0.98(0.96-0.99),respectively;the SENp, SPEp, +LRp, -LRp, DOR and AUROC in F0-F2 vs F3-F4 group were 92.9%(88.9%-95.5%),94.6%(91.2%-96.8%),17.348(10.496-28.671),0.075(0.048-0.119),230.434(111.482-476.317)0.98(0.96-0.99), respectively;the SENp, SPEp, +LRp,-LRp, DOR and AUROC in F0-F3 vs F4 group were 97.7%(93.0%-99.3%),93.2%(90.3%-95.2%),14.337(9.910-20.742),0.025(0.008-0.075),580.405(144.871-2325.307) and 0.98(0.96-0.99),respectively.Conclusion:MRE,as a new and noninvasive imaging method, has high diagnostic value in all stages of hepatic fibrosis, which can provide a reliable reference for clinical precise treatment of hepatic fibrosis.

Silencing ATM gene gave rise to enhanced apoptotic response to irradiation and irradiation-like chemotherapy agents, this paper explored the crucial identities of the molecular elements responsible for the enhanced apoptotic response in U937 cells mediated by silencing ATM gene. Two U937 cell mutants named U937-ASPI3K (ATM, negative) and U937-pZeosv2(+) (ATM, wild-type) were used as a cell model system to identify the critical molecule(s) responsible for the varied apoptotic response in the absence or presence of ATM gene. Apoptosis was examined by measuring concentrations of free nucleosome in U937 cells. Western blot was employed to measure nuclear protein abundance of CDC25A, CDC25B, CDC25C, total p34cdc2, p34cdc2, (Thr 161) or p34cdc2 (Thr 14, Tyr 15). RT-PCR was used to estimate CDC25 transcript levels. U937-ASPI3K exhibited an enhanced apoptotic response to lower dosage of irradiation, which could not be blocked by protein synthesis inhibitor. Protein serine-threonine phosphatase inhibitor or cyclin-dependent kinase (CDK) inhibitors, on the other hand, abolished the enhancement indicated that protein phosphorylation/dephosphorylation modification and CDK activity are required for the enhanced apoptotic response in the absence of ATM gene. Upon irradiation, p34cdc2 in U937-pZeosv2(+) was maintained in an inactive state by phosphorylation on threonine 14 (Thr 14) and tyrosine 15 (Tyr 15), which was associated with a dramatic decrease of nuclear CDC25A, CDC25B and CDC25C proteins. In contrast, p34cdc2 in U937-ASPI3K maintained in an active state by dephosphorylation on threonine 14 (Thr 14) and tyrosine 15 (Tyr 15), which was associated with constant nuclear CDC25A, CDC25B and CDC25C protein abundance before and after irradiation. The responsive decrease of nuclear CDC25 proteins occurred at the post-transcription level. Silencing ATM gene blocks the responsive decrease of nuclear CDC25 proteins, which is responsible for failure to inactivate p34cdc2 after irradiation. Active p34cdc2 and CDK2, in turn, acts as the death executors to trigger apoptosis. In summary, aberrantly activated CDK activity is the critical molecular mechanism central to enhanced apoptotic responses in the absence of ATM gene.

Objective To provide scientific basis for developing mental interventions for the wounded in major disasters by assessing the mental health status and the mental intervention effect among the Wenchuan earthquake survivors transferred to Chongqing.Methods A total of 389 transferred wounded survivors were examined by using questionnaires including general information questionnaire,Psychological Health Self-rating Questionnaire (PHSQ),Symptom Checklist 90 (SCL-90),Perceived Social Support Scale (PSSS) and Coping Styles Questionnaire (CSQ).According to PHSQ,the wounded were divided into control group and stress group.Then,the stress group was given mental intervention and taken the second mental evaluation one month later.Results Acute stress disorder (ASD) rate (30.08％) was remarkably observed in these transferred wounded,who were subsequently administered a set of mental intervention.The wounded whose PHSQ mark ≥8 were defined as the stress group (Group A) and the others as the control group ( Group B).Compared with Group B,Group A had higher scores in depression ( P ＜ 0.01 ),anxiety ( P ＜ 0.01 ) and consternation ( P ＜ 0.01 ),and lower scores in PSSS (P ＜0.01 ).Group A inclined to negative coping styles mainly including shrink back (P ＜ 0.01 ) and fantasy (P ＜0.01 ),while Group B inclined to coping styles mainly including rationalization and help.The PSSS score in Group A was significant lower than that in Group B (P ＜ 0.01 ).The scores in PHSQ,SCL-90 (anxiety and consternation),CSQ and PSSS were significantly improved after the mental intervention in Group A.Conclusions The incidence of ASD is high in the transferred wounded survivors and is mainly related with mental stress,such as emotional symptoms,negative coping styles and low perception of social support.Early mental interventions will help improve the psychological status of the transferred wounded.

Objective To investigate the characteristics of the mental health and the change of serum copeptin,cortisol and aldosterone levels of the soldiers in field army under military exercises stress.Methods A total of 294 soldiers were evaluated with the psychological stress self evaluation test (PSET) before the exercises and the soldiers whose PSET score were T ＜ 70 had to accept the survey with the general information questionnaire before the exercises, and another two questionnaires including PSET and Symptom Checklist 90 (SCL-90) after the exercises, the higher stress group and lower stress group were defined according to the PSET score, T≥70 and T ＜ 70 respectively.Meanwhile, we draw the venous blood of them in order to detect the serum concentration of copeptin,cortisol and aldosterone before and after the military exercise.Results ( 1 ) After the survey of PSET before the exercises,263 soldiers were determined to be the final research objects.According to the score of PSET after the exercises, 104 soldiers(39.5% ) were considered as the higher stress group,and the symptoms mainly included anxiety,hostility and poor sleep and appetite.(2) Compared with the lower stress group,the higher stress group were younger(21.51 ± 3.13 vs 24.09 ± 4.16, P ＜ 0.01 ), higher at the proportion of soldiers ( 68.27% vs 54.09%, P＜0.01 ) and lower at the proportion of soldiers who have experiences of military exercises(37.50% vs 57.23%, P＜0.01 ).(3) Compared with the lower stress group,the higher stress group had higher score in EPQN ( 10.43 ± 1.78 vs 8.77 ± 1.67, P ＜ 0.01 ).(4) The serum levels of copeptin ( P ＜ 0.01 ), cortisol ( P ＜ 0.01 )and aldosterone( P＜0.01 ) have increased after the military exercise.Compared with the lower stress group, the higher stress group have higher serum copeptin level ( 11.36 ± 2.21 vs 9.43 ± 2.34, P ＜ 0.01 ) after the exercises.( 5 ) The relationship between PSET score and the copeptin level after exercise can be expressed as a formula: PSET score = 27.909 + 3.669X ( X: copeptin level after exercise), and 11.472 pmol/L was considered as the threshold value.Conclusion As a strong stress agent, military exercises may cause obvious physiological changes and has notable effect on related hormones.The change of serum copeptin level can be used as a sensitive indicator to determine the degree of stress.