Objective To establish a specific HPLC method and a standard fingerprint for quality control ofZiziphi Spinosae Folium.Methods The samples were separated on a Kinetex C18 column (100 mm × 2.1 mm,2.6 μm) by gradient elution at the flow rate of 300 μL/min using acetonitrile and 0.1％ aqueous formic acid as the mobile phase.The column temperature was 30℃.The detection wavelength was 225 nm and the sample size was 10 μL.The fingerprint evaluation software (2012 edition) for Chinese materia medica (CMM) was used to evaluate the similarity of the 12 batches of samples.Results There were 13 characteristic peaks identified in the characteristic spectra ofZiziphi Spinosae Folium samples.Peak 6 was Rutin.The similarities of 13 batches ofZiziphi Spinosae Folium samples were proved to behigher than 0.850.Conclusion The method is available with a good reproducibility and accuraty which can control the quality standards effectively.

BACKGROUND:Periprosthetic joint infection is a complication that is difficult to deal with after joint arthroplasty. Early diagnosis is the key to treatment. To find a fast response, high-sensitivity and high-specificity molecular biomarker can significantly optimize the diagnosis process of periprosthetic joint infection. OBJECTIVE:To monitor blood procalcitonin, interleukin-6 and lipopolysaccharide binding protein levels, to compare with blood leukocyte count and C-reactive protein levels, to identify above indexes, and to distinguish sensitivity and specificity of periprosthetic joint infection. METHODS: A total of 81 patients with pain after arthroplasty who were treated in Affiliated Hospital of Jining Medical Colege from January 2008 to December 2013 were enroled in this study. The repair surgery of al patients was divided into two stages. In the first stage, complete debridement and the instalation of temporary occupancy device were conducted. After 3 months averagely, two-phase reconstruction was performed. At 1 day before surgery, venous blood was colected. Calcitonin, interleukin 6, lipopolysaccharide binding protein, leukocyte count and C- reactive protein levels were detected. During the operation, synovial membrane and sample of false envelope around the prosthesis were colected. Bacterial and histological examinations were performed. The sensitivity and specificity were calculated using receiver operating characteristic curve.
RESULTS AND CONCLUSION: One-way analysis of variance results showed that the receiver operating characteristic curve of lipopolysaccharide binding protein was bigger, 0.962; 95 confidence interval 0.924-1.000. Diagnostic value was optimal, and the critical value was 23.5 μg/L. These data suggested that when lipopolysaccharide binding protein exceeded 23.5 μg/L before surgery, periprosthetic joint infection would be identified. The receiver operating characteristic curve of C-reactive protein was 0.871. The receiver operating characteristic curve of leukocytes was close to 0.5. The diagnostic value of leukocyte count on periprosthetic joint infection was not great. These findings indicate that lipopolysaccharide binding protein has good application prospect in the diagnosis of periprosthetic joint infection after joint replacement, and shows high positive predictive rate and negative predictive rate of periprosthetic joint infection.

Objective To study the tooth germ development of mouse molar in fluorosis.Methods We established an acute fluorosis model and used HE-staining after preparing specimens of different stage of mouse first molar developing tooth germ.Results The developing molars of the control fetuses were at the bell stage of odontogenesis,whereas those of the experimental population were at cap stage.The ameloblasts became shorter and lost polarity.The arrangement of ameloblasts fell into disorder at the bell of differeiational stage and secretory stage.Conclusion The developing molars of the experimental fetuses were retarded.High-dose fluoride has a strong effect in the ameloblasts at the bell of differetiational stage and secretory stage.

Objective:To explore the effects of miR-181a-5p on the occurrence and development of gastrointestinal stromal tumors (GIST) through targeting CTDSPL mediating TGF-β signaling pathway.Methods:Surgical treatment of GIST patients in the First People’s Hospital of Shangqiu City from Jan. 2016 to Dec. 2019 were selected as research objects, and tumor tissue and adjacent normal tissue were collected intraoperatively. The clinicopathological data of the patients were analyzed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression of CTDSPL gene and miR-181a-5p expression. Western blot was used to detect the protein level of CTDSPL and TGF-β signaling pathway related factors. Human gastrointestinal stromal tumor cell lines (GIST-T1) were transfected with miR-181a-5p mimic, miR-181a-5p inhibitor, or CTDSPL overexpression vector. MTT was used to detect cell proliferation activity, Transwell assay was utilized to detect cell invasion, flow cytometry was used to determine cell apoptosis in each group.Results:Compared with adjacent tissues, expression of miR-181a-5p and TGF-β signaling pathway related factors was activated while CTDSPL expression was inhibited. Tumor size, invasion depth and modified NIH grading were related to the mRNA expression level of CTDSPL gene in GIST tumor tissues (All P<0.05) . Compared with Blank group, inhibition of miR-181a-5p or CTDSPL overexpression had the ability to inhibit the cell viability and invasion, induce apoptosis. The effects of miR-181a-5p mimic on GIST-T1 can be saved by CTDSPL overexpression. Conclusion:miR-181a-5p can promote the occurrence and development of GIST by down-regulating the CTDSPL gene level and activating TGF-β signaling pathway.

Objective To explore the effects of cAMP response element binding protein ( CREB) on taxol-induced cell cycle arrest in HeLa cells .Methods MTT assay was used to determine the optimal concentration and treatment time . PCR method was used to construct the recombinant plasmid pCI neo /CREB( PN) and site-directed mutagenesis recombinant plasmid pCI neo/CREB-M(PM).Cell cycle was assayed by flow cytometry .Expressions of pCREB, CREB, cyclins and CDKs were assayed by Western blotting .Results The effective conditions of taxol treatment on HeLa cells were 0.1μmol/L for 24 hours.After cells were treated with 0.1μmol/L taxol, G2/M phase was arrested in a time-dependent manner , accomplished with the decrease of cyclin A , a significant increase of cyclin B1, D1 and phosphorylated CREB (pCREB) protein expression, whereas, no marked changes were observed in cyclin E , CDK1, CDK2, CDK4 and CREB expressions. However, combinantion of PM and taxol treatment significantly reduced taxol-induced G2/M phase arrest, and reversed the effect of taxol-decreased cyclin A, increased cyclin B1 and D1 expression.Conclusion Tanscription factor CREB-mediated specific cyclins play a pivotal role in taxol-induced G 2/M arrest in HeLa cells .

Objective:To summarize the clinical features and experience in diagnosis and treatment of subglottic cysts in children.Methods:The clinical data of 5 children with subglottic cysts admitted in Qilu Children′s Hospital of Shandong University from April 2018 to June 2019 were retrospectively analyzed.Results:Among the 5 patients, 2 cases were premature infants and 3 cases were full term infants.All patients had an endotracheal intubation history.One patient was asymptomatic.Four patients had laryngeal stridor, 2 cases of whom were accompanied by hoarseness and dyspnea.These 2 patients were used to be misdiagnosed with laryngomalacia, laryngitis, etc.Bronchoscopy revealed subglottic gray-white cyst-like lesions, and enhanced CT showed low density without enhancement.All patients were treated with laser and forceps, and bronchoscope was applied in the follow up period for 5-12 weeks.There was no recurrence or subglottic stenosis.Conclusions:Laryngeal stridor in children with a history of endotracheal intubation must be examined by bronchoscopy in time to find out whether there is subglottic cysts.Laser therapy is an effective treatment for subglottic cysts.

The clinical data of 4 children with congenital absence of tracheal cartilage ring in Qilu Children′s Hospital of Shandong University from November 2017 to August 2019 were retrospectively analyzed.The age of the 4 cases was from 2 months to 1 year.One case had no obvious symptoms after birth, and then had repeated wheezing attacks for 3 times.Another 3 cases had symptoms after birth, such as throat ringing, repeated cough and asthma.Bronchoscopy in 4 cases indicated that the local lumen of trachea was round and centripetal stenosis, but the body of bronchoscope could pass smoothly, and no cartilage structure was found in the wall of trachea.Among them, 3 cases of chest enhanced CT examination displayed local and hourglass-like tracheal stenosis, without vascular ring malformation.All the 4 cases were treated by operation, of which 3 cases recovered well and 1 case died of severe infection after operation.

Objective:Colorectal cancer is one of the common gastrointestinal tumors.Recent studies have shown that, the expression of PDEF can promote the differentiation of Secretory progenitor cells to goblet cells in the intestinal tissue.Therefore the oc-currence of colorectal cancer may be related to expression of PDEF.In this study,we tried to investigate the effects of proliferation and invasion after interference targeting prostate-derived ETS factor in colorectal cell lines HT29.Methods: HT29 cells were transiently transfected with PDEF shRNA plasmids and blank control plasmid via cathodolyte liposome transfection method.By fluorescence microscopy,RT-PCR,Western blot technique to detect the expression of PDEF mRNA and protein in normal control group,blank control group,shRNA group.The proliferation and invasion ability of HT29 cells after transfection were assessed by MTT assay and Transwell invasion assay respectively.Results: Green fluorescent protein was observed in blank control plasmid group and shRNA plasmid group.Western blot showed the reduced PDEF protein expression compared with normal control group and blank control group.Interference PDEF gene expression can significantly promote the proliferation of HT29 cells (P<0.05).The ability of cell invasion in interference group was significantly higher than the normal control group and blank control group after 48h ( P<0.05).Conclusion:Interference PDEF in HT29 cells can promote cell proliferation and invasion.

BACKGROUND:Cytokines and neurotrophic factors secreted from human umbilical cord blood-derived mesenchymal stem cells secrete have neuroprotective effects on cerebral ischemia-reperfusion injury, but there are few reports about intranasal administration of human umbilical cord blood-derived mesenchymal stem cellconditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the protective effects of intranasal administration of human umbilical cord-derived mesenchymal stem cells-conditioned medium on neurologic function of rats with cerebral ischemia-reperfusion injury.
METHODS:Adult rats were subjected to 2 hours of right middle cerebral artery occlusion and the human umbilical cord-derived mesenchymal stem cells were isolated from the postpartum human cord. We made the conditioned medium of human umbilical cord-derived mesenchymal stem cells. Ischemic rats were randomized and assigned to three groups and were treated by intranasal routine starting 24 hours after middle cerebral artery occlusion with:(1) saline for control group;(2) Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium for medium control group;(3) conditioned medium treatment group (10mL/kg) daily for 14 days. Behavioral tests (foot fault test, and modified Neurological Severity Score) were performed before and at 1, 7, 14 days after middle cerebral artery occlusion.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperatively 1 day (P>0.05). Compared to the control and medium control group rats, respectively, rats in the conditioned medium group significantly improved functional outcome after stroke in days 7 and 14 (P<0.05). There was also no significant difference in functional tests between the control group and medium control group in days 7 and 14 (P>0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells-conditioned medium via intranasal administration can significantly improve neurologic functional outcome after cerebral ischemia-reperfusion injury.

Objective To establish and compare HPLC fingerprint chromatograms of Dipsaci Radix decoction pieces, aqueous decoction and formula granules.Methods The HPLC analysis was carried out in Wondasil C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of acetonitrile-0.1% phosphoric acid by gradient elution. The flow rate was 1.0 mL/min; the detection wavelength was set at 212 nm; the column temperature was kept at 30℃. Results The fingerprint chromatograms from 12 batches of Dipsaci Radix decoction pieces, aqueous decoction and formula granules were established respectively. 14 common peaks in the fingerprint chromatogram in the formula granules could be tracked in the aqueous decoction, and 13 common peaks in the fingerprint chromatogram could be tracked in the decoction pieces. 2 chemical compounds were identified, such as asperosaponinⅥ and chlorogenic acid.ConclusionThe method of HPLC fingerprint chromatograms is stable and with good repeatability. Dipsaci Radix decoction pieces, aqueous decoction and formula granules are basically the same chemical composition.