Objective To discuss the clinical value and nursing of the three-endoscope in the treatment of eholedoeholithiasis. Methods 45 eases of choledocholithiasis patients who were treated with LCDE (three-endoscope) were named as the research group.56 patients who received traditional open ab-dominal surgery were set as the control group. The average hospitalization time and satisfaction degree with nursing were compared, t test and χ2 test were adopted. Results The average hospitalization time was shorter and satisfaction degree with nursing was higher in the research group than those in the control group. Conclusions The treatment of choledochohthiasis with three-endoscope is safe and feasible, es-pecially when combined with antibiotics lavage and stone dissolution through naso-biliary duct.The opera-tion can widen the surgical indication,reduce the risk of surgery with little damage,clear stones completely, reduce postoperative complicatioas,make patients recover faster, shorten the hospital stay and achieve the same or better treatment results when compared to the traditional open abdominal surgery.

Objective:To analyze deceleration capacity of heart rate (DC) in patients with ventricular arrhythmia , and explore its predictive value for ventricular arrhythmia .Methods :Dynamic electrocardiogram (DCG) data of outpatients and inpatients ,who were treated in our hospital from Jan 2012 to Jul 2015 ,were retrospectively ana‐lyzed .A total of 41 rehabilitation patients with ventricular tachycardia (VT) directly detected by DCG were treated as VT group ,another 37 cases with similar general data and non‐tachycardia were enrolled as non‐tachycardia group in the same period .DCG was used to measure and calculate DC in all subjects , DC indexes were compared between two groups .Results:DC<4.5 ms was regarded as abnormal and DC<2.5 ms was regarded as significantly abnor‐mal .Compared with non-tachycardia group ,DC significantly reduced [ (8.72 ± 1.78) ms vs .(4.01 ± 1.90) ms] , P<0.01 ;there were significant rise in abnormal DC rate (5.41% vs .51.22% ) and significant abnormal DC rate (0 vs .29.27% ) in VT group , P<0.01 both .Conclusion:Detection rate of abnormal DC is high in VT patients .DC measurement is helpful for predicting ventricular arrhythmia and sudden cardiac death .

BACKGROUND: How to establish a stable in vitro culture system, including location of corneal limbal epithelial stem cells, in vitro sample harvest, in vitro culture, vector selection, as well as identification methods, play a key role in corneal limbal epithelial stem cells culture. OBJECTIVE: To culture the isolated rabbit corneal limbal epithelial stem cells and to identify the biological properties of cultured cells. METHODS: The primary rabbit cornel limbal epithelial stem cells were isolated and cultured with tissue inoculation using human amniotic membrane as vector. The growth features of cells were observed under an inverted microscope. The morphology of cells was observed by hematoxylin-eosin staining and a scanning electron microscope. Furthermore, the monoclonal antibody AE5 and P63 two-step immunohistochemical staining were used to identify limbal epithelial stem cell protein expression. RESULTS AND CONCLUSION: The rabbit corneal limbal epithelial stem cells could be successfully cultured and maintained a relatively high value-added potential in vitro. Rabbit corneal limbal epithelial stem cells cultured on the amniotic membrane pull netted cellular layer. The AE5 monoclonal antibody positive rate of primary cultured cells was about 5% and P63 monoclonal antibody positive up to 90%. AE5-positive rate increased and P63-positive rate decreased with the increase in the number of subculture. The rabbit limbal epithelial stem cells can be successful culture and amplified on human amniotic membrane in vitro by limbal tissue culture method. The cultured cells maintain the characteristics of corneal epithelial cells. The rabbit corneal limbal epithelial stem cells can form grafts on the amniotic membrane.

OBJECTIVE: Explore the model of universal NICU newborns' hearing screening in high-risk neonates, preliminary understanding factor of hearing damage. METHOD: Transient evoked otoacoustic emissions (TEOAE) and automatic auditory brainstem response (AABR) were used to detect newborns' hearing in 13 315 objects, that is newborns' hearing screening in NICU with TEOAE test who not pass, 42 days after will use AABR rescreening. Children's Hearing Center of Guangxi Child Health Hospital will diagnose the newborns that did not pass in 3 months. RESULT: In these 13 315 newborns, 5 151 subjects who did not pass the initial screening, 1910 subjects who also did not pass after 42 days, 1167 subjects cannot pass the rescreening after 3 months, 642 subjects were diagnosed congenital hearing impairment by Brainstem Auditory Evoked Potential Test, the rate is 4.82%. CONCLUSION TEOAE and AABR are the suitable model of universal newborns' hearing screening in high-risk neonates.
Evoked Potentials, Auditory, Brain Stem ; Female ; Follow-Up Studies ; Hearing Tests ; Humans ; Infant, Newborn ; Intensive Care Units, Neonatal ; Male ; Neonatal Screening

OBJECTIVETo explore a convenient and effective method for norovirus nucleic acid extraction from oysters suitable for long-term viral surveillance.

METHODSTwo methods, namely method A (glycine washing and polyethylene glycol precipitation of the virus followed by silica gel centrifugal column) and method B (protease K digestion followed by application of paramagnetic silicon) were compared for their performance in norovirus nucleic acid extraction from oysters. Real-time RT-PCR was used to detect norovirus in naturally infected oysters and in oysters with induced infection.

RESULTSThe two methods yielded comparable positive detection rates for the samples, but the recovery rate of the virus was higher with method B than with method A.

CONCLUSIONMethod B is a more convenient and rapid method for norovirus nucleic acid extraction from oysters and suitable for long-term surveillance of norovirus.

Animals ; Centrifugation ; methods ; Norovirus ; genetics ; isolation & purification ; Ostreidae ; virology ; RNA, Viral ; isolation & purification

Objective:To investigate the mechanism of Wenjing Decoction in the treatment of hepatic fibrosis through network pharmacological methods, and conducting animal experiments to verify the core targets.Methods:The TCMSP database platform was used to screen the active components and related targets of Wenjing Decoction, and the Uniprot database was used to obtain the target genes corresponding to the active components of Wenjing Decoction. The network diagram of "Chinese materia medica-compound-target" was constructed in Cytoscape 3.7.2, and the GeneCards database was used to search liver fibrosis related targets. String database was used to construct a protein interaction network (PPI) to screen the core components and key targets of liver fibrosis, and GO analysis and KEGG pathway enrichment were performed. Animal experiments were conducted to verify the results of the analysis. 10 mice were selected as the blank group, and the remaining 45 rats were induced with carbon tetrachloride induced liver fibrosis model. After modeling, 40 successfully modeled rats were divided into model group and Wenjing Decoction high, medium-, and low-dosage groups using a random number table method, with 10 rats in each group. Wenjing Decoction high, medium-, and low-dosage groups were orally administered with 1.5×3.18, 3.18, and 0.5×3.18 g/kg Wenjing Decoction, respectively. The blank group was orally administered with equal volume distilled water once a day for 8 consecutive weeks. HE staining was used to observe the histopathological and morphological changes in the liver of rats. The serum GPT and GOT levels of rats were detected using a fully automated biochemical analyzer, and the expressions of TNF, AKT, and IL-6 proteins in rat liver tissue was detected using Western Blot.Results:A total of 188 active components of Wenjing Decoction were obtained, and the active components with higher degree values were β-sitosterol, quercetin, naringenin, etc. 799 liver fibrosis gene targets were collected, and the core target genes of the PPI network were TNF, AKT, IL6, etc. The key anti-hepatic fibrosis related pathways were obtained by GO function and KEGG analysis, including pathway in cancer, TNF, PI3K-Akt and other signalling pathways. Results of animal experiments showed that there were obvious inflammatory infiltration, collagen fibre and pseudo lobe generation in the liver tissue of rats in the model group, and the levels of inflammation and fibrosis in the liver tissue of rats in the Wenjing Decoction high, medium-, and low-dosage groups were improved to different degrees compared with that of the model group; compared with the model group, the levels of serum GPT and GOT decreased ( P<0.05); the protein expressions of TNF, AKT and IL6 in the Wenjing Decoction high, medium-, and low-dosage groups decreased ( P<0.05). Conclusion:Wenjing Decoction may exert anti-liver fibrosis effects by intervening in TNF, AKT, IL6 targets, regulating cancer pathways, TNF, PI3K Akt and other signaling pathways.

Objective To explore the effect of first-line anti-tuberculosis treatment on vitamin D level in patients with pulmonary tuberculosis,and to master the changes of vitamin D level in the course of treatment,so as to provide a scientific basis for tuberculosis and nutrition health education in Shenzhen.Methods A total of 100 patients diagnosed as smear-positive pulmonary tuberculosis and receiving initial treatment in 2016 were enrolled and all the patients were treated with the standardized short-course chemotherapy regimens.The blood samples were extracted before treatment and at the ends of intensive and continuation phase.The 25-hydroxyvitamin D [25-(OH) D] concentrations were determined by chemiluminescence (CLIA) at each time point.The change of 25-(OH) D concentrations during anti-tuberculosis treatment was analyzed and the differences of vitamin D levels between different time points were identified.Results 79 (79.0％),94 (94.0％) and 96 (96.0％) patients were found vitamin D deficiency before treatment and at the end of the intensive and continuation phases respectively,which showed an upward trend (x2=15.543,P＜0.001) and the 25-(OH)D concentrations were (15.74±6.54) ng/ml,(12.56±5.15) ng/ml,(11.51±4.28) ng/ml,respectively.During the whole course of treatment,the 25-(OH) D concentration decreased by 26.9％ or (4.23 ± 6.75) ng/ml (t =6.257,P＜0.001),wherein it decreased (3.18 ± 5.24) ng/ml in intensive phase (t =6.069,P＜ 0.001) and (1.05±4.86) ng/ml in continuation phase (t =2.154,P =0.034).The former had a greater decreased value (t=2.836,P=0.006).There were 77 (77.0％) and 55 (55.0％) patients with 25-(OH)D concentration reduction in intensive and continuation phases respectively (x2 =9.680,P =0.003),of which 41 patients (41.0％) continued to decline.Conclusion Once anti-tuberculosis treatment is conducted,the vitamin D level will decrease rapidly in the intensive phase and continue decreasing throughout the course of treatment,which leads to a general lack of vitamin D in patients with primary pulmonary tuberculosis.First-line anti-tuberculosis drugs may be the main cause for vitamin D level reduction.Therefore,it is necessary for clinicians to strengthen vitamin D health education for each patient throughout the treatment period,especially for those at high risk of vitamin D deficiency who should be recommended adjuvant vitamin D supplementation therapy.

Objective To prepare a dissolvable microneedle(DMN)with a tip-layer loaded with hyaluronic acid(HA)modified sinomenine hydrochloride liposomes(HA-SMH-Lip),as well as characterize,evaluate its in vitro transdermal permeability,cellular uptake ability,and anti-inflammatory ability.Methods HA-SMH-Lip-DMNs were prepared by a two-step casting method,and the drug loading capacity was determined using HPLC.The morphology,skin permeation properties and in vitro transdermal ability were investigated by scanning electron microscopy,puncture assay and Franz diffusion cell method.Fluorescent microneedles were prepared by replacing HA-SMH-Lip with fluorescein isothiocyanate liposomes(HA-FITC-Lip/FITC-Lip).The uptake behavior of inflammation cells on HA-FITC-Lip-DMNs/FITC-Lip-DMNs was investigated using a flow cytometer and a fluorescence microscope.To evaluate the anti-inflammatory activity of HA-SMH-Lip-DMNs,the levels of inflammatory factors including nitric oxide(NO),tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β),and IL-10 in cell supernatants were measured using an ELISA kit.Results The prepared HA-SMH-Lip-DMNs have uniform shape and size,integral and visually pleasing array,and an average drug loading of(114.01±1.04)μg.Additionally,they have good puncture ability.The results of in vitro transdermal experiments showed that the accumulated amounts of HA-SMH-Lip-DMNs were(101.47±2.91)μg·cm-2 at 36 hours.Its transdermal ability was better than that of the SMH solution group and SMH liposome group.In vitro cellular uptake results indicated that HA-FITC-Lip-DMNs were more effectively taken up by RAW 264.7 cells(P＜0.01).Compared to the model group,HA-SMH-Lip-DMNs group significantly reduced TNF-α,IL-1β,and NO levels while increase IL-10 levels(P＜0.01).Conclusion The prepared HA-SMH-Lip-DMNs have a complete and beautiful morphology with excellent cellular uptake capability,remarkable in vitro transdermal performance,and potent anti-inflammatory properties.HA-SMH-Lip-DMNs are expected to become a new type of transdermal drug delivery system.

OBJECTIVE To establish the fingerprints of Tianma jiannao granules （TJG） and the method for content determination to evaluate the quality of TJG comprehensively combined with chemometric analysis. METHODS High-performance liquid chromatography （HPLC） was used to establish the fingerprints of 13 batches （S1-S3） of TJG and determine the contents of inosine， gastrodin， parishin B and parishin E. Cluster analysis， principal component analysis， and orthogonal partial least squares- discriminant analysis were performed using SPSS 20.0 and SIMCA 18 software； using variable importance projection （VIP） value greater than 1 as a criterion， marker components that affected quality were screened. RESULTS A total of 28 common peaks were identified in the 13 batches of TJG with similarities greater than 0.9， and 7 common peaks were identified， which were gastrodin， p-hydroxybenzyl alcohol， parishin B， parishin E， rhynchophylline， inosine and salidroside. The 13 batches of TJG were clustered into 3 categories， S1-S2， S8-S10 and S12 were clustered into one category； S3 and S7 were clustered into one category； S4-S6， S11 and S13 were clustered into one category. VIP of inosine was greater than 1. The contents of inosine， gastrodin， parishin B and parishin E were 62.637-176.677， 17.821-37.642， 5.748-16.077 and 5.660-13.510 μg/g. CONCLUSIONS The established HPLC fingerprints and content determination method are stable， reliable and highly reproducible， which can be used to evaluate the quality of TJG in combination with chemometric analysis. Inosine may be a marker component that affects the quality of TJG. There are differences in the quality of 13 batches of TJG.

Objective To observe the changes and significance of the protein expression levels of nuclear factor-κB p65 (NF-κB p65), transforming long factor-β1 (TGF-β1) and apoptosis-related factors Bcl-2 and Bax in myocardial tissue of Bama miniature pig model of diabetic cardiomyopathy (DCM). Methods Ten healthy male Guangxi Bama miniature pigs, aged 4 to 5 months old, were selected and divided into control group and model group according to the random number table method, with 5 pigs in each group. After 12 hours of fasting in the two groups, the DCM model was replicated by intravenous injection of streptozotocin (STZ) 150 mg/kg; for the Bama miniature pigs in the control group, citric acid-sodium citrate buffer 150 mg/kg was injected intravenously. After 10 months of modeling, the basic conditions of the two groups of animals were observed and their fasting blood glucose (FPG) levels were detected. The protein expression levels of NF-κB p65, TGF-β1, Bcl-2 and Bax in myocardial tissue of two groups were detected by Western Blot and the pathological changes of myocardial tissue were observed under electron microscope. Results In the model group, 4 models were successfully established, and 1 died. The model pigs had symptoms such as polydipsia, polyphagia, polyuria and decreased body weight. The FPG level in the model group was significantly higher than that in the control group (mmol/L: 25.53±3.75 vs. 4.68±0.77, P < 0.01). Compared with the control group, the protein expression levels of NF-κB p65, Bax and TGF-β1 in the myocardial tissue of model group were significantly increased (NF-κB p65/GAPDH: 0.46±0.05 vs. 0.38±0.02, Bax/GAPDH: 0.46±0.01 vs. 0.35±0.01, TGF-β1/GAPDH: 0.39±0.01 vs. 0.33±0.01, all P < 0.05) and the expression level of Bcl-2 protein was significantly decreased (Bcl-2/GAPDH: 0.33±0.01 vs. 0.42±0.01, P < 0.01). Electron microscopy results showed that the myofibrils of myocardial tissue in the DCM model group were disordered, and the number of mitochondria in the gap was significantly reduced. A large number of mitochondria with vacuolar degeneration were observed. Conclusions The DCM model of Bama miniature pigs can be successfully replicated after 10 months of high-dose STZ disposable ear vein injection. The DCM model miniature pigs have obvious glucose metabolism disorder, and their myocardial tissue has inflammatory reaction, cardiomyocyte apoptosis and fibrosis.