Human with bi-allelic WNT10A mutations and epithelial Wnt10a knockout mice present enlarged pulp chamber and apical displacement of the root furcation of multi-rooted teeth,known as taurodontism;thus,indicating the critical role of Wnt10a in tooth root morphogenesis.However,the endogenous mechanism by which epithelial Wnt10a regulates Hertwig's epithelial root sheath(HERS)cellular behaviors and contributes to root furcation patterning remains unclear.In this study,we found that HERS in the presumptive root furcating region failed to elongate at an appropriate horizontal level in K14-Cre;Wnt10afl/flmice from post-natal day 0.5(PN0.5)to PN4.5.EdU assays and immunofluorescent staining of cyclin D1 revealed significantly decreased proliferation activity of inner enamel epithelial(IEE)cells of HERS in K14-Cre;Wnt10afl/flmice at PN2.5 and PN3.5.Immunofluorescent staining of E-Cadherin and acetyl-α-Tubulin demonstrated that the IEE cells of HERS tended to divide perpendicularly to the horizontal plane,which impaired the horizontal extension of HERS in the presumptive root furcating region of K14-Cre;Wnt10afl/flmice.RNA-seq and immunofluorescence showed that the expressions of Jag1 and Notch2 were downregulated in IEE cells of HERS in K14-Cre;Wnt10afl/fl mice.Furthermore,after activation of Notch signaling in K14-Cre;Wnt10afl/flmolars by Notch2 adenovirus and kidney capsule grafts,the root furcation defect was partially rescued.Taken together,our study demonstrates that an epithelial Wnt10a-Notch signaling axis is crucial for modulating HERS cell proper proliferation and horizontal-oriented division during tooth root furcation morphogenesis.

Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of tobramycin in human serum,and examine the utility of the method in a clinical pharmacokinetic study of tobramycin inhalation.Methods Serum samples were pretreated by solid phase extraction with tobramycin-D12 as internal standard.Chromatographic separation was performed on a TitankHilic(2.1 mm × 100 mm,3 μm)column.The mobile phase consisted of0.1％formic acid-acetonitrile and 0.1％formic acid aqueous solution at a flow rate of 0.4 mL/min.Electrospray ionization source and multiple reaction monitoring(MRM)scanning were used for monitoring the quantitative ion pairs with m/z 468.3→m/z 163.3(tobramycin)and m/z 480.6→m/z 166.2(tobramycin-D12).The established method was investigated in terms of selectivity,interaction,concomitant medication,standard curve and lower limit of quantitation,precision and accuracy,recovery,matrix effect,and stability of tobramycinin.Results The linear range of tobramycin was 0.050 0-10.0 mg/L(R2=0.999 5).The intra-and inter-batch precision was satisfactory(coefficient of variation[CV]≤3.6％).The accuracy ranged from-0.4％to 6.0％.The matrix effect factor(MF)in human serum samples(including hemolysis and lipemia)ranged from 92.2％to 94.9％(CV≤2.7％).The recovery of tobramycinin was 79.5％-81.9％in serum samples,while the recovery of internal standard was 78.9％.The analyte was stable in serum samples for 72 h at room temperature and for 274 days at-20℃/-70℃.The pharmacokinetic study of tobramycin inhalation in bronchiectasis patients showed that after continuous administration of tobramycin 300 mg twice a day to 3 patients,the mean Cmax of tobramycin was(0.72±0.61)mg/L on Day 1 and(0.76±0.73)mg/L on Day 28,respectively.The corresponding Tmax was(1.83±0.61)h and(1.50±0.50)h,respectively.Conclusions The UPLC-MS/MS method established in this study is sensitive,accurate and rapid.It is successfully applied to the clinical pharmacokinetic study of tobramycin inhalation.The method may be suitable for therapeutic drug monitoring of tobramycin in clinical practice.

CaMKII is essential for long-term potentiation (LTP), a process in which synaptic strength is increased following the acquisition of information. Among the four CaMKII isoforms, γCaMKII is the one that mediates the LTP of excitatory synapses onto inhibitory interneurons (LTP_E→I). However, the molecular mechanism underlying how γCaMKII mediates LTP_E→I remains unclear. Here, we show that γCaMKII is highly enriched in cultured hippocampal inhibitory interneurons and opts to be activated by higher stimulating frequencies in the 10-30 Hz range. Following stimulation, γCaMKII is translocated to the synapse and becomes co-localized with the postsynaptic protein PSD-95. Knocking down γCaMKII prevents the chemical LTP-induced phosphorylation and trafficking of AMPA receptors (AMPARs) in putative inhibitory interneurons, which are restored by overexpression of γCaMKII but not its kinase-dead form. Taken together, these data suggest that γCaMKII decodes NMDAR-mediated signaling and in turn regulates AMPARs for expressing LTP in inhibitory interneurons.
Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Hippocampus/metabolism* ; Interneurons/physiology* ; Long-Term Potentiation/physiology* ; N-Methylaspartate/metabolism* ; Receptors, AMPA/physiology* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Synapses/physiology*

Immediate-early genes (IEGs) have long been used to visualize neural activations induced by sensory and behavioral stimuli. Recent advances in imaging techniques have made it possible to use endogenous IEG signals to visualize and discriminate neural ensembles activated by multiple stimuli, and to map whole-brain-scale neural activation at single-neuron resolution. In addition, a collection of IEG-dependent molecular tools has been developed that can be used to complement the labeling of endogenous IEG genes and, especially, to manipulate activated neural ensembles in order to reveal the circuits and mechanisms underlying different behaviors. Here, we review these techniques and tools in terms of their utility in studying functional neural circuits. In addition, we provide an experimental strategy to measure the signal-to-noise ratio of IEG-dependent molecular tools, for evaluating their suitability for investigating relevant circuits and behaviors.
Animals ; Brain ; metabolism ; Gene Expression Profiling ; methods ; Genes, Immediate-Early ; Humans ; Molecular Imaging ; methods ; Neural Pathways ; metabolism ; Neurons ; metabolism ; Signal-To-Noise Ratio

Objective? To explore the application effect of multi-form training assessment in clinical training of geriatric nursing specialties. Methods? By convenience sampling, a total of 34 nurses under training from 14 hospitals in Beijing were selected in December 2018, and divided into experimental group(n=17) and control group(n=17) randomly. The control group adopted the traditional teaching method. A practical demand-oriented training course based on the characteristics of geriatric nursing specialties was established, and various forms of training were used, multi-station assessment of the training effect was conducted in the experimental group. The training lasted for one month in both groups. The assessment scores and satisfaction degree were compared between the groups. Results? In the experimental group, the student nurses＇ elderly patient management test score (95.0±7.5), the elderly patients specialist nursing test station test score(97.0±6.7), and the emergency response test station test score (99.0±4.5) were all higher than the control group with (86.0±8.8), (80.0±10.5), (88.0±7.6) respectively, and the differences were statistically significant (P＜0.01); the difference between the two groups of student nurses in satisfaction degree towards trainers was not statistically significant (P＞0.05); there were statistical differences between the two groups in teaching curriculum, teaching method, teaching contents, and assessment forms in both of the groups (P＜ 0.01); the overall satisfaction degree was 97.6%(83/85) in the experimental group, higher than the control group with 62.3%(53/85) and the difference was statistically significant (χ2=38.594,P＜0.001). Conclusions? Multi-form training is suitable for teaching nurses in clinical geriatric specialty. It can improve the test scores and learning satisfaction of the student nurses. Scientific curriculum, multi-form teaching methods and multi-station assessment are the key in the improvement of training quality.

OBJECTIVE: To explore the prognostic factors of patients with unresectable liver metastasis colorectal cancer after failed conversion chemotherapy. METHODS: A retrospective, case-controlled study was performed. Study subjects were 105 patients who were diagnosed with synchronous liver metastasis colorectal cancer after failed chemotherapy (metastasis evaluated as unresectable after the conversion chemotherapy) at Xinhua Hospital, Shanghai Jiaotong University from January 2012 to December 2015. Overall survival(OS) was retrospectively analyzed using Kaplan-Meier method. Log-rank test was used to compare survival among groups. Univariate and multivariate analysis was conducted for prognosis using Cox regression model. RESULTS: Of 105 cases,70 were male and 35 were female with median age of 60 years old. Twenty-one patients had right colon cancer, 41 had left colon cancer, 42 had rectal cancer and 1 had synchronous cancers(sigmoid colon and rectum). One hundred and two (97.1%) patients were cT3-4 and 90 patients were cN+ (imaging diagnosis). Eighty-nine (84.8%) patients were loaded with 2 or more liver metastases with the median maximum diameter of 48.3 mm. The patients were followed up for 3 to 43 months from the day of diagnosis. The median OS was 11 months (interquartile range, 8-18). The median OS of patients with cN0, cN1 and cN2 stage was 17, 13 and 10 months, respectively(P=0.026). The median OS of patients with single lesion, 2-3 lesions, 4-10 lesions and more than 10 lesions was 15, 15, 17 and 9 months, respectively (P=0.002). OS of patients with maximum diameter of liver metastatic lesion ≤ 50 mm, 51-100 mm and >100 mm was 15, 10 and 8 months, respectively(P=0.003). The median OS of patients with chemotherapy response of partial response (PR), stable disease (SD) and progressive disease (PD) was 17, 14 and 8 months, respectively(P<0.001). OS was 17 months in patients receiving second line chemotherapy and was 10 months in those without second line chemotherapy (P<0.001). OS in patients undergoing primary tumor resection was 13 month and in those without primary tumor resection was 9 months; the difference was not significant (P=0.060). Multivariate analysis showed that cN2(HR=2.115, 95%CI:1.089-4.109, P=0.027), the maximum diameter of liver metastatic lesion more than 100 mm (HR=3.112, 95%CI:1.455-6.657, P=0.003), chemotherapy response of PD (HR=4.435, 95%CI:2.506-7.533,P<0.001) and without second line chemotherapy(HR=4.432,95%CI:2.186-8.986, P=0.010) were independent prognostic factors. CONCLUSIONS For patients with unresectable liver metastasis from colorectal cancer after failed conversion chemotherapy, prognostic factors include cN2, the maximum diameter of liver metastatic lesion, chemotherapy response and second line chemotherapy. Whether the resection of primary tumor can prolong OS further study.
Antineoplastic Agents ; therapeutic use ; China ; Colorectal Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Liver Neoplasms ; diagnosis ; secondary ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Failure

Objective To evaluate the effect of Babaodan capsules combined with transcatheter arterial chemoembolization (TACE) therapy in patients with BCLC stage B primary hepatocellular carcinoma (HCC). Methods A total of 32 patients with BCLC B primary hepatocellular carcinoma who cannot be treated with operation in the Department of Oncology in Fujian Province Cancer Hospital from May 2010 to March 2013 were enrolled in the experimental group. According to paired design, 64 patients with BCLC B primary liver cancer who were treated with only TACE were as control. The patients of experimental group were treated with Babaodan capsules combined with TACE, and the patients of control group were treated with only TACE. Overall response rate (ORR), time to progression (TTP), overall survival (OS), liver function change 1 week after TACE, and post-embolization syndromes were analyzed. Measurement data were compared using t test, count data were compared with χ2test. Kaplan-Meier method was used for survival analysis, and Log-rank method for testing. Results The ORR 1.5 month after TACE was 75.0%in experimental group and 81.3%in control group(P=0.477). The median TTP was 8.9 months(95%CI 3.1-14.7 months) in experimental group, and 5.5 months (95%CI 4.3-6.7 months) in control group (P=0.048). The median OS time was 16 months(95%CI 8.0-24.0 months) in experimental group, and 12 months(95%CI 11.0-13.0 months) in control group (P=0.159). Compared with the experimental group, the alanine transaminase 1 week after TACE in control group increased obviously (P=0.018). The incidence rate of ≥CTCAE grade 2 pain after the first time TACE in experimental group was lower than that in control group(P=0.019). Conclusion Babaodan capsules could reduce pain of HCC patients after TACE, improve liver damage after TACE,and prolong the TTP of patients with BCLC stage B HCC.

Objective To investigate the safety , feasibility , and clinical application value of digital sub-traction angiography ( DSA) -guided insertion of totally implantable venous access port ( TIVAP) in patients with digestive system cancer .Methods We retrospectively analyzed the data of 15 digestive system cancer patients who were inserted with TIVAP under DSA guidance between April 2013 and January 2016, recorded the TIVAP-related complications and indwelling time, and investigated the patients’satisfaction about TIVAP.Paired rank sum test was used for the differences in patients’quality of life before and after the insertion of TIVAP.Results Of the 15 digestive system cancer patients , operation success rate of TIVAP insertion was 100%, with the success rate of venipuncture on first attempt being 100%.The incidence of complication was 6.67% ( 1/15 ) , which was manifested as pull feeling at the neck occurring in the eighth month after insertion .The indwelling time of TIVAP was from 2 to 28 months, with a median value of 9 months.Patients’satisfaction rate of TIVAP was 86.67%(13/15).The proportion of patients with a good quality of life was 100%(15/15), statically signifi-cantly higher than that before the insertion [46.67%(7/15), Z=-3.416, P=0.000).Conclusions TIVAP insertion under DSA guidance in digestive system cancer patients is safe and feasible , with few complications and fair patient satisfaction .It may improve the patient ’s quality of life , worthy of clinical application .

OBJECTIVETo evaluate the effect of annular electrode lacrimal duct reconstruction in improving the safety and efficacy of nasolacrimal duct stent implantation for treatment of nasolacrimal duct obstruction.

METHODSThis randomized clinical trial was performed to compare the efficacy, success rate of intubation, time used for stent implantation, intraoperative pain, and extubation-assciated complications between nasolacrimal stent implantation with and without annular electrode lacrimal duct reconstruction.

RESULTSA total of 119 eligible patients were enrolled in this trial. The total curative rate at 6 months of follow up after extubation was 70.9% (83/117) in these patients, and was significnatly higher in pateinets with lacrimal duct reconstruction than in those without [80.6% (54/67) vs 58.0% (29/50); χ(2)=7.093, P<0.05]. The total success rate of stent implantation was 98.3% (117/119) in all the patients initially enrolled, and two patients experienced failure of stent implantation and were excluded; the success rate was signfiicantly higher in patients initially enrolled in the lacrimal duct reconstruction group (χ(2)=6.282, P<0.05). The median time required for intubation was shorter in lacrimal duct reconstruction group (12 s vs 33 s; Z=-36.722, P<0.05). The intendity of intraoperative pain was comparable between the two groups (t=0.833, P=0.405). The total rate of puncta injury was 43.6% (51/117) in these patients and similar between the two groups (χ(2)=1.459, P=0.227). The total rate of extubation difficulty was 9.4% (11/117) in all the patients, and was lower in lacrimal duct reconstruction group [4.5% (3/67) vs 16% (8/50); χ(2)=4.463, P<0.05]. Stent breakage in extubation occurred in 4.3% (11/117) of the patients with similar rates between the two groups (χ(2)=2.964, P=0.085). Spearman bivariate correlation analysis showed that the time required for intubation was inversely correlated with the treatment efficacy (r=-0.584, P<0.05) and positively with the occurrence of extubation difficulty (r=0.491, P<0.05); extubation difficulty was inversely correlated with the curative effect (r=-0.511, P<0.05).

CONCLUSIONAnnular electrode nasolacrimal duct reconstruction can increase the safety and efficacy of nasolacrimal duct stent implantation for treatment of nasolacrimal duct obstruction.

Dacryocystorhinostomy ; methods ; Electrodes ; Humans ; Intubation ; Lacrimal Duct Obstruction ; Nasolacrimal Duct ; surgery ; Pain ; Reconstructive Surgical Procedures ; methods ; Stents ; Treatment Outcome

The body is exposed to various organic anions,so it is the best way to remove the toxic substances in the body quickly and effectively.Cross epithelial active transport mediated by organic anion transporter is the rate limiting process.Renal secretion and re-absorption of a variety of endogenous and exogenous organic anions are occurred in the proximal tubular epithelial cells of the organic anion transporter family.The expression of organic anion transporter-1 (OAT1) in proximal tubular epithelial cells plays an important role in the introduction of organic anion into the renal tubular epithelial cells.This article reviewed the renal expression,the substrate and the polymorphism of the organic anion transport protein OAT1,the renal toxicity of adefovir dipivoxil,the interaction between organic anion transporters and drug,and the influence of the renal toxicity on the renal toxicity of adefovir dipivoxil.