Objective To investigate the effect of dopaminergic neurons of midbrain ventral tegmental area(VTA) in general anesthesia.Methods Forty adult healthy male SD rats were randomly divided into lesion group (n=20) and control group (n=20).The lesion group was given the bilateral infusion of specific dopaminergic neuron injury agent 6-OHDA in midbrain lateral VTA,while the control group received the same volume of normal saline at the same areas.The time of loss of righting reflex (LORR)loss and recovery of righting reflex(RORR)at postoperative 2 week were observed in each group.Results Compared with the control group,the LORR time in the lesion group was shortened and the RORR time was significantly prolonged under propofol-induced anesthesia (P＜0.05).However,the LORR time under the isoflurane anesthesia had no statistically significant difference between the two groups(P＞0.05),while the RORR time in the lesion group was increased(P＜0.05).Conclusion Dopaminergic neurons in midbrain VTA might play different roles in the induction and recovery of different general anesthetics.

Objective To determine the function of conserved amino acids in the fusion-promoting domain of Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein,clearly understanding mechanism of cell fusion.MethodsUsing a PCR-based site-directed mutagenesis method and the method of homology recombination occurred in vivo to change six conservative amino acids into alanine respectively.Wild type (WT) and all mutant HN proteins were exepressed in BHK-21 cells by the vacciniaT7 RNA polymerase expression system.The amount of each HN protein at the cell surface was determined by fluorescence-activated cell sorter (FACS).Cell fusion efficiency,hemadsorption activity (or receptor binding activity) and neuraminidase activity were determined.Results There was no statistic difference of cell surface expression among WT and each mutant HN protein ( P＜0.05 ).Cell fusion efficiency of each mutant protein decreased to some extent,especially 1103A decreased to 14.2％ in head.Hemadsorption activity of mutant proteins were reduced in different extent,the maximum reduction of which was also 1103A,28.2％ of wt NDV HN.There was different neuraminidase activity among each mutant HN protein.L74A increased slightly to 118.6％.L110A decreased most to 5.2％.I103A decreased second most to 5.7％.Conclusion Conserved amino acids in fusion-promoting domain of NDV HN played an important role in cell fusion.I103 was identified as a key amino acid in this domain.

ObjectiveTo study the effect of sub-chronic exposure to deltamethrin(DM) on neural behavior and expression of NMDA receptor in hippocampus of mice.Methods 60 Female SPF Kunming mice were divided into 4 groups and given DM 60 days by gavage.Hot-plate,rotarod,grip strength,hing limb landing foot splay were used to examine the sensory and motor change of mice.Autonomic activity test was used for detecting the functional status of the central nervous system in mice.Passive avoidance test for detection of the behavior changes of learning and memory,and RQ-PCR was employed to measure the expression of NMDA receptor in hippocampus of mice.ResultsThe behavior of sensory and motor of mice sub-chronic exposure to deltamethrin did not have changes significantly(P ＞ 0.05 ).In the test of autonomic activity test,the average of autonomic activity times were (93 ± 18) times,(107 ± 13) times,(105 ±22) times.Compared with the control group,the average of autonomic activity times in middle-and high-dose groups were increased significantly (P ＜ 0.05 ).The latent periods in poisoning groups were (175.4 ±38.4) s,(146.4 ±51.2)s,(132.3 ±45.0) s,and the error times were (0.7 ±0.3)times,( 1.4 ± 0.5 ) times,( 1.8 ± 0.9) times.Compared with the control group,latent periods of high-dose group were decreased and the error times of middle-and high-dose groups were increased significantly (P ＜ 0.05 ).Compared with the control group,the relative expression levels of NR1 and NR2A mRNA in hippocampus of middle and high-dose groups were increased significantly (P＜ 0.05 ),and the relative expression level of NR2B mRNA in highdose group was decreased significantly(P ＜ 0.05).ConclusionSub-chronic exposure to DM could increase the excitability of mice,damage the function of learning and memory,and influence the expression of NMDA receptor in hippocampus of mice.

Objective To construct a chimeric infectious clone of the fatal virulent strain SDLY 107, containing the gene fragments encoding 2A and 3B proteins of the mild virulent strain SDLY 1, and to establish a reverse genetic system platform for further investigation on virulence of enterovirus 71 strains. Methods The overlap PCR analysis was performed to obtain the gene fragments encoding 2A and 3B pro-teins of the mild virulent strain SDLY 1.The obtained gene fragments were digested and then cloned into a plasmid pMD19-T containing the full-length gene of SDLY 107 strain by using gene replacement strategy. The recombinant RNA was transfected into Vero cells for the preparation of recombinant virus particles.Sev-eral assays including the PCR, indirect immunofluorescence ( IFA) , Western blot and sequencing were per-formed for virus identification.Virus titers were measured by 50%cell culture infective dose ( CCID50 ) and plaque assay.Results The infectious clones of SDLY 107-2A-1 and SDLY 107-3B-1 chimeric virus strains were constructed successfully.Typical cytopathic effect was observed in Vero cells after viral transfection. Identification of the rescued viruses by PCR, IFA, Western blot and sequencing further confirmed the suc-cessful construction of infectious virus strains.The virus titers of SDLY 107-2A-1 and SDLY 107-3B-1 strains detected by CCID50 and plaque assay were 1.25 ×105 PFU/ml and 0.7 ×105 PFU/ml, respectively. Conclusion The chimeric viruses SDLY 107-2A-1 and SDLY 107-3B-1 were rescued successfully, causing cytopathic effects similar to those by using the parental virus strain SDLY 107.This study might pave the way for further investigation on in vitro and in vivo virulence of enterovirus 71 strains.

Dens invaginatus is a rare developmental anomaly of the teeth that is caused by the infolding of enamel organs or the penetration of their proliferations into dental papillae before calcification has occurred. The presence of double dens invaginatus is extremely rare. This paper describes the use of cone beam computed tomography in the evaluation of a maxillary lateral incisor with double dens invaginatus and periapical periodontitis. The tooth was treated through microscopic root canal therapy. The tooth was free of clinical symptoms, and the periradicular lesion narrowed during the follow-up period of 1 year.
Humans ; Dental Pulp Cavity/abnormalities* ; Dens in Dente/pathology* ; Incisor/pathology* ; Root Canal Therapy ; Periapical Periodontitis/pathology*

Objective: To explore the effect of enteral nutrition on tumor cell proliferation activity in rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy. Methods: Sixty-six rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy from January 2016 to January 2018 in the Yongchuan Hospital Affiliated to Chongqing Medical University were selected. The patients were divided into experimental group (enteral nutrition combined with neoadjuvant therapy) and control group (simple adjuvant therapy) according to the random digits table method, with 33 cases in each group. The expressions of proliferating cell nuclear antigen (PCNA) and Ki-67 antigen before and after treatment were detected by immunohistochemical method; the albumin and prealbumin before and after treatment were observed, and the nutrition risk screening 2002 (NRS2002) was evaluated. Results: There were no statistical differences in the expressions of PCNA and Ki-67 antigen before treatment between 2 groups (P>0.05); the expressions of PCNA and Ki-67 antigen after treatment in experimental group were significantly lower than those in control group, and there were statistical differences (P<0.05). There were no statistical differences in the NRS2002 score, albumin and prealbumin before treatment between 2 groups (P>0.05); the NRS2002 score after treatment in experimental group was significantly lower than that in control group: (1.58 ± 0.50) scores vs. (3.65 ± 0.72) scores, the albumin and prealbumin after treatment were significantly higher than those in control group: (35.92 ± 2.77) g/L vs. (31.12 ± 1.76) g/L and (204.58 ± 23.86) mg/L vs. (157.46 ± 18.99) mg/L, and there were statistical differences (P<0.01). Conclusions Enteral nutrition can reduce the proliferation activity of tumor cell in rectal cancer patients with nutritional risk treated with preoperative neoadjuvant therapy, and it can improve the nutritional status of patients.

Objective:To investigate the clinical outcome after surgery and first 131I treatment in patients with moderate-risk papillary thyroid cancer (PTC), and analyze the relevant factors that affect the therapeutic effect. Methods:From January 2018 to April 2019, 135 patients (48 males, 87 females; age (42.7±11.1) years) with moderate-risk PTC in the Second Affiliated Hospital of Chongqing Medical University were retrospectively analyzed. According to the 2015 American Thyroid Association (ATA) guidelines, patients were divided into excellent response (ER) group, inderteriminate response (IDR) group, biochemical incomplete response (BIR) group and structural incomplete response (SIR) group, of which IDR, BIR, SIR were collectively referred to as the non-ER group. χ2 test and Mann-Whitney U test were used to compare the general clinical features between the ER and non-ER groups, and then multivariate logistic regression analysis was performed. The predicted value of pre-ablation stimulated thyroglobulin (ps-Tg) to ER was assessed by ROC curve analysis. Results:The treatment responses of 94 patients were ER, and those of 41 were non-ER. The differences in tumor size (0.80(0.50, 1.10) vs 1.00(0.55, 1.50) cm; U=1 491.50, P=0.036), the number of metastatic lymph nodes (3(2, 5) vs 4(2, 12); U=1 422.00, P=0.015), metastatic lymph node size (0.50(0.30, 0.65) vs 0.50(0.30, 1.45) cm; U=1 396.50, P=0.013), metastatic lymph node involvement rate (50%(30%, 70%) vs 60%(50%, 85%); U=1 441.50, P=0.024), metastatic lymph node location (central/lateral: 76/18 vs 24/17; χ2=7.40, P=0.007) and ps-Tg level (2.1(0.8, 5.3) vs 14.0(3.2, 35.2) μg/L; U=680.00, P<0.001) were statistically significant between the ER and non-ER groups. Multivariate logistic regression analysis showed that ps-Tg (odds ratio ( OR)=1.200, 95% CI: 1.107-1.302, P<0.001) was an independent factor influencing ER. The cut-off value of ps-Tg for predicting ER was 7.38 μg/L, with the sensitivity and specificity of 68.3%(28/41) and 87.2%(82/94) respectively. Conclusion:Moderate-risk PTC patients with smaller tumor size, fewer metastatic lymph nodes, lower metastatic lymph node involvement rate, metastatic lymph nodes in central area, smaller metastatic lymph node size, and ps-Tg<7.38 μg/L have better therapeutic effect after initial 131I treatment.

Objective To compare the production of farnesol between Candida albicans (C.albicans) biofilms formed by resistant and standard strains in different media,and to investigate the changing trend of farnesol production in different phases of biofilm formation and the features of farnesol production by resistant C.albicans.Methods Fluconazole-resistant C.albicans strains were induced in vitro.Standard strains and fluconazole-resistant strains of C.albicans were separately inoculated onto different media,including RPMI 1640 medium,yeast extract peptone dextrose (YPD) medium,yeast nitrogen base (YNB) + 0.5％ glucose medium,RPMI 1640 + 10％ fetal calf serum (FCS),so as to form C.albicans biofilms.Morphological changes of C.albicans biofilms at 24 hours were observed under an inverted microscope,and gas chromatography-mass spectrometry (GC/MS)was performed to detect the level of farnesol at 1.5,3,6,12,24,36 and 48 hours.Results There were no obvious differences in the morphology of C.albicans biofilms between the resistant and standard strains when they were cultured in the same medium,while the morphology of C.albicans biofilms markedly differed between the 2 kinds of strains in the different media.Three-factor analysis of variance showed that the production of farnesol in the C.albicans biofilms changed over time (F =70.628,P ＜ 0.001).Concretely speaking,during the formation of resistant and standard C.albicans biofilms,the production of farnesol gradually increased in the RPMI 1640,YPD and YNB + 0.5％ glucose media until the biofilms matured,then showed a decreasing trend.However,the time to peak levels of farnesol was different between the 2 kinds of strains in these media.Moreover,the levels of farnesol in the 2 kinds of strains both slowly increased in the RPMI 1640 + 10％ FCS medium within 12-48 hours.Culture media also significantly affected the production of farnesol (F =176.665,P ＜ 0.001),and the levels of farnesol in the resistant and standard C.albicans biofilms were both higher in the YNB + 0.5％ glucose medium.When resistant and standard strains were separately cultured in the RPMI 1640 media and the YPD media,the level of farnesol was significantly higher in the resistant strains than in the standard stains (RPMI 1640 media at 36 hours:1.157 ± 0.064 vs.0.250 ± 0.075,P ＜ 0.05;YPD media at 6 hours:0.262 ± 0.036 vs.0.055 ± 0.062,P ＜ 0.05;YPD media at 12 hours:0.730 ± 0.030 vs.0.482 ± 0.024,P ＜ 0.05).However,when they were separately cultured in the YNB + 0.5％ glucose media,the farnesol level was significantly higher in the standard stains than in the resistant strains (36 hours:2.950 ± 0.677 vs.0.523 ± 0.266,P =0.020).Conclusion The media markedly affect the production of farnesol in the C.albicans biofilms,and there is a certain difference in the production of farnesol between resistant and standard C.albicans strains.

Metal-organic frameworks (MOFs), comprised of organic ligands and metal ions/metal clusters

Sepsis-induced uncontrolled systemic inflammatory response syndrome (SIRS) is a critical cause of multiple organ failure. Acute kidney injury (AKI) is one of the most serious complications associated with an extremely high mortality rate in SIRS, and it lacked simple, safe, and effective treatment strategies. Leontopodium leontopodioides (Willd.) Beauv (LLB) is commonly used in traditional Chinese medicine for the treatment of acute and chronic nephritis. However, it remains unclear whether lipopolysaccharide (LPS) affects LPS-induced AKI. To identify the molecular mechanisms of LLB in LPS-induced HK-2 cells and mice, LLB was prepared by extraction with 70% methanol, while a lipopolysaccharide (LPS)-induced HK-2 cell model and an AKI model were established in this study. Renal histopathology staining was performed to observe the morphology changes. The cell supernatant and kidney tissues were collected for determining the levels of inflammatory factors and protein expression by ELISA, immunofluorescence, and Western blot. The results indicated that LLB significantly reduced the expression of IL-6 and TNF-α in LPS-induced HK-2 cells, as well as the secretion of IL-6, TNF-α, and IL-1β in the supernatant. The same results were observed in LPS-induced AKI serum. Further studies revealed that LLB remarkably improved oxidative stress and apoptosis based on the content of MDA, SOD, and CAT in serum and TUNEL staining results. Notably, LLB significantly reduced the mortality due to LPS infection. Renal histopathology staining results supported these results. Furthermore, immunofluorescence and Western blot results confirmed that LLB significantly reduced the expression of the protein related to the NF-κB signaling pathway and NLRP3, ASC, and Caspase-1 which were significantly increased through LPS stimulation. These findings clearly demonstrated the potential use of LLB in the treatment of AKI and the crucial role of the NF-κB/NLRP3 pathway in the process through which LLB attenuates AKI induced by LPS.
Animals ; Mice ; NF-kappa B/metabolism* ; Lipopolysaccharides/adverse effects* ; NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Acute Kidney Injury/metabolism* ; Kidney ; Systemic Inflammatory Response Syndrome/pathology*