OBJECTIVE To investigate the mechanism through which Danggui buxue decoction regulates neutrophil extracellular traps （NETs） to improve osteoporosis （OP） in rats with premature ovarian failure （POF）. METHODS Female SD rats were randomly divided into normal group， model group， calcitriol group， and Danggui buxue decoction low-dose， medium-dose and high-dose groups， with 9 rats in each group. Except for the normal group， all other groups were administered cisplatin via intraperitoneal injection on days 1 and 8 to establish a POF complicated with OP model. Each group received the corresponding drugs or normal saline intragastrically starting from day 5， once a day， for 4 consecutive weeks. After the last medication， serum levels of estradiol （E2）， NETs， 25-hydroxyvitamin D3 [25（OH）D3]， receptor activator of nuclear factor-κB ligand （RANKL）， and osteocalcin （BGP） were measured. The histopathological changes in bone tissue were observed. The expressions of vitamin D receptor （VDR）， myeloperoxidase （MPO）， neutrophil elastase （NE） and citrullinated histone H3 （CitH3） in bone tissue were detected； the protein expressions of 25-hydroxyvitamin D-1α-hydroxylase （CYP27B1） and 1α，25-dihydroxyvitamin D3-24-hydroxylase （CYP24A1） were also determined. RESULTS Compared with the normal group， the bone tissue of rats in the model group showed a significant reduction in the number of trabeculae， which was thinner broken and poorly connected， with significant destruction of the reticular structure， and an enlarged marrow cavity. Serum levels of NETs and RANKL， the protein expressions of MPO， NE， CitH3 and CYP24A1 in bone tissue were significantly increased or upregulated， while serum levels of E2， 25（OH）D3 and BGP as well as protein expressions of VDR and CYP27B1 were significantly decreased or downregulated （P＜0.05）. Compared with the model group， the histopathological changes in the bone tissue of rats in each administration group showed some degree of recovery， with significant improvements observed in most quantitative indicators （P＜0.05）. CONCLUSIONS Danggui buxue decoction can restore the E2 level in POF complicated with OP rats， and improve OP. The mechanism may be related to its ability to upregulate VD level and inhibit the formation of NETs.

As an important means of data visualization, statistical graphics can potentially convey scientific findings and key points. However, authoritative graphic guidelines are lacking currently, and medical researchers still have shortcomings in mastering graphic design principles and drawing skills, which may obscure some key research findings and even misinterpret the research findings. Based on statistical examples, we summarized the essential elements and design principles of statistical graphics to address these challenges and to provide a reference for medical researchers.

To explore the remineralization effect of bioactive glass NovaMin on demineralized dentin specimens, and to study the physical and chemical properties of formed structure at dentin surface.
 Methods: One mm-thickness coronal dentin slices were soaked in ethylene diamine tetraacetic acid (EDTA) for 48 h to prepare the completely demineralized dentin specimens and they were divided into 2 groups: an artificial saliva group (control group) and a NovaMin powder group. The specimens were treated with artificial saliva or NovaMin powder for 2 min (2 times every day), and the interval was 8 hours. Then, the specimens were soaked in the remineralization solution. After 7 days, the scanning electron microscope (SEM), energy dispersive X-ray (EDX), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray diffraction (XRD) were used to detect dentin morphology, the physical and chemical properties of the formed structure at dentin surface.
 Results: The results of SEM showed that a defined surface layer in the NovaMin powder group could be observed in the SEM imaging at the 7th day, which completely occluded dentinal tubules; the EDX, ATR-FTIR and XRD analysis found that the mineralized layer formed at dentin surface was mainly composed of calcium and phosphate elements, which was similar to the hydroxyapatite-like crystal. However, there were no materials formed at the dentin surface in the control group, and the dentinal tubules were still open.
 Conclusion: NovaMin can remineralize the demineralized dentin specimens and occlude the dentinal tubules in hydroxyapatite-like crystal structure.
Dental Pulp Cavity ; Dentin ; chemistry ; Glass ; Microscopy, Electron, Scanning ; Saliva, Artificial ; Spectroscopy, Fourier Transform Infrared ; Tooth Remineralization ; methods

AIM:To investigate the role of Rho-associated kinase ( ROCK) inhibitor fasudil in the formation of rabbit urethral stricture after injury and to observe the cell activity , migration and extracellular matrix synthesis in the rabbit urethra fibroblasts.METHODS:The rabbit model of urethral stricture was established by microsurgical techniques .The rabbits were divided into sham operation group , operation group and fasudil (3 mg/kg, 10 mg/kg, 30 mg/kg) groups.The diameter of the stenosis was measured by retrograde urethrography 3 months after surgery .The fibroblasts were isolated from urethral scar, and then incubated with fasudil (12.5 μmol/L, 25 μmol/L, 50 μmol/L) in the presence of transforming growth factor-β1 (TGF-β1, 10 μg/L).The untreated cells were used for control .The cell activity was measured by MTT assay.The cell migration ability was tested by the method of Transwell chambers .The protein expression of ROCK , α-smooth muscle actin (α-SMA) , collagen I and collagen III was determined by Western blot analysis .RESULTS:Fasudil significantly reduced formation of urethral stricture after injury (P<0.05).Cultured rabbit fibroblasts with different con-centrations of fasudil inhibited the cell activity and cell migration ability (P<0.05).The protein expression of ROCK,α-SMA, collagen I and collagen III was also inhibited by treatment with fasudil in a dose -dependent manner ( P<0.05 ) . CONCLUSION:Fasudil inhibits the formation of extracellular matrix and reduces the incidence of urethral stricture after injury by down-regulating TGF-β1-induced Rho/ROCK pathway activation in the rabbit urethra fibroblasts .

Objective To establish a method for fingerprint analysis of amino acids from honey by high performance liquid chromatography ( HPLC). Methods Amino acids of honey were concentrated by 732 cation exchange resin, and then were treated by pre-column derivatization with phenyl isothiocyanate, with praline as control peak. The chromatography was performed on a Waters Symmetry C18 ( 250 mm × 4.6 mm × 5 μm) column, with acetonitrile ∶ water (4∶1) as mobile phase A and 30 mmol/L sodium acetate ∶ acetonitrile (355∶15, acetic acid adjusting pH value to be 6.5) as mobile phase B by gradient elution. The detection wave length was set at 254 nm. The flow rate was 1.0 mL/min. The column temperature was 40℃, and the injection volume was 5μL. Results Sixteen common peaks were shown in the fingerprint of 15 batches of honey samples. The similarity for 15 batches of honey samples was in the range of 0.910 ~ 0.996 . Conclusion The fingerprint detection method is simple, practical, reproducible and specific, and can provide certain reference for quality control of honey.

Objective To understand the nosocomial infection distribution,drug resistance characteristics and carbapenemases-producing phenotype of Pseudomonas aeruginosa (PAE)and Acinetobacter baumannii (ABA).Methods The nosocomial infection strains of non-repeated PAE and ABA isolated in this hospital and the infected cases from July 2012 to July 2013 were retrospec-tively collected.The antimicrobial susceptibility test was conducted by the disk diffusion method(K-B method).The modified Hodge test was adopted to preliminarily screen carbapenemase and the positive strains of preliminary screening were further detected met-allo-beta-lactamase(MBL)by 2-mercaptopropionic acid synergy test.Results During the study period,250 strains of non-repeated PAE and 132 strains of ABA were included.All of them were primarily isolated from sputum specimens,accounting for 55.5%.The department distribution was dominated by the intensive care units(ICU),accounting for 20.9%.The antimicrobial susceptibility test showed that the sensitivity of PAE to the testing anti-microbial drugs was more than 70%,its resistance rates to IPM and MEM were 8.5% and 9.5% respectively.However,the resistance rates of ABA to the testing anti-microbial drugs were up to 35.2%-77.4%,its resistance rates to IPM and MEM were 35.2%,39.1% respectively.The occurrence rates of multidrug-resist-ant and pandrug-resistant ABA nosocomial infection was higher than that of PAE,which were 44.7% and 24.0% and 9.1% and 2.8%,respectively.Among 40 strains of carbapenem-resistant PAE,11 strains(27.5%)were positive in the preliminary screening and 2 strains(18.2%)were positive of MBL phenotype.Among 49 strains of carbapeneme-resistant ABA,37 strains(75.5%)were positive in the preliminary screening and only 1 strain(2.7%)was positive of MBL phenotype.Conclusion PAE and ABA in our hospital exhibit different resistance to common antibacterial drugs.The monitoring should be strengthened.The production of car-bapenemsa is one the main mechanisms for PAE resistance to carbapenems.The detection rate of MBL-producing PAE and ABA is lower in our hospital.

Objective To explore the optimum dilution concentration of calcium gluconate intravenous infusion in infants and the detaining time of venous indwelling needle .Methods One hundred and thirty-nine infants who needed calcium gluconate intravenous infusion were chosen and divided into the one time dilution group (n=43), two times dilution group (n=48), and five times dilution group (n=48).The incidence of extravasation injury in puncture vascular , the degree of injury and the situation of healing were compared among three groups .Results The total incidence of extravasation injury in puncture vascular in the one time dilution group, two times dilution group , five times dilution group within 3 days were respectively 19.4%, 9.0%, 2.1%, and the differences were statistically significant (H=261.23, P<0.01).The further analysis showed that the incidence of extravasation injury in the one time dilution group was higher than that of two times dilution group (χ2 =6.88, P<0.01), and the incidence in the two times dilution group was higher than that of five times dilution group (χ2 =6.40, P<0.05), and the incidence in the one time dilution group was higher than that of five times dilution group (χ2 =20.48, P <0.01 ).No difference was found in the incidence of extravasation injury in puncture on the first , second and third day of the detaining time of venous indwelling needle (H=3.47,P>0.05).Conclusions The incidence of extravasation injury is lowest in 5 times diluted concentration of 10%calcium gluconate intravenous infusion in infants through venous indwelling needle .

Objective To analyze the prevalence status and the genetic characterizations of influenza B viruses isolated in Hunan Province after pandemic influenza A (H1N1) 2009,and to explore possible reasons for the prevalence.MethodsThroat swabs were collected from outpatients with influenza-like illness in 23 sentinel hospitals of Hunan Province in 2010.Influenza viruses were isolated with Madin-Darby canine kidney (MDCK) cells and identified by haemagglutination inhibition test.The genomes of 10 selected influenza B viruses were sequenced and analyzed for phylogenetic and molecular characterization.ResultsWith the reduction of isolation of pandemic influenza A (H1N1)2009 viruses,influenza B virus became the predominant isolated strain in the first half of 2010.Epidemic viruses mainly belonged to the B/Victoria lineage,and both two lineages co-circulated.Seven out of 11 influenza outbreaks caused by type B.Ten strains were filled into 2 branches of BV and BY which were classified by their lineage types in polymerase (PB2,PB1,PA),hemagglutinin (HA),neuraminidase (NA),NB,membrane protein (M1),influenza B virus membrane protein M2 (BM2),and non-structural protein (NS1,NS2) phylogenetic trees except the NP phylogenetic tree in which 10 strains were all in the BY branch.Compared with World Health Organization (WHO) vaccine strains,the amino acid identity of 11 proteins of the 10 strains was high (97.2％-100.0％).However,some amino acid point mutations were found.No mutation was found in drug resistance mutation sites.Some mutations in NA,NB,PB1,PB2 and NS2 molecules were found in 2 strains isolated from outbreaks compared with strains from sentinel surveillance.Conclusions The point mutations,insertions and genetic reassortment indicate viruses sustaining evolution,which is probably the reason for predominant influenza B viruses after pandemic influenza A (H1N1) 2009 in Hunan Province.

Objective To evaluate the effect of enteral nutrition (EN) versus total parenteral nutrition(TPN) on gut barrier function in patients with severe acute pancreatitis (SAP). Methods Sixtythree patients with SAP enrolled from 4 hospitals were randomly assigned into EN group(29 cases) and TPN group(34 cases). EN group patients were fed via a spiral nasojejunal feeding tube placed routinely by endoscopy or fluoroscopy, and TPN group patients were nourished intravenously with TPN during the same period. The changes of serum endotoxin, diamine oxidase, and urinary excretion of lactulose and mannitol ratio (L/M) were observed. Results Plasma concentration of endotoxin were markedly decreased in EN group as compared with that in TPN group at the 7th,14th ,21th day of entry trial [(39. 30 ± 15. 82) EU/L vs (73.05 ±21.16) EU/L,(22.64 ±14.31) EU/L vs (49.34 ±24.54) EU/L,(14.81 ± 10.93)EU/L vs ( 30. 08 ± 14. 10 ) EU/L, P ＜ 0. 05]. Plasma concentration of diamine oxidase were markedly decreased in EN group as compared with that in TPN group at the 7th, 14th day of entry trial [(9. 97 ± 3. 84)U/Lvs (19.89±9.89)U/L,(5.42±1. 84) U/Lvs (8.79 ±4.08) U/L, both P ＜ 0. 05]. The urinary L/M decreased significantly in EN group than those in TPN group at the 7th, 14th,21th day of entry trial (0.28 ±0.25 vs 0. 65 ±0.45,0.21 ±0. 18 vs 0.54 ±0.41,0.08 ±0.04 vs 0.29 ±0.06, all P＜0.05).Conclusion EN has better effect on improving intestinal barrier function than TPN in treatment of patients with SAP.

OBJECTIVETo investigate the chemical constituents of Peperomia tetraphylla.

METHODThe constituents of EtOAc-soluble portion were isolated and purified by chromatography. Their structures were identified by spectral features.

RESULTSix amides were isolated and identified as, aristololactam All (1), aristololactam B II (2), N-trans-feruloyltyramine (3), N-trans-sinapoyltyramine (4), N-trans-feruloylmethoxytyramine (5), N-p-coumaroyltyramine (6).

CONCLUSIONAll compounds were isolated from this plant for the first time.