The abuse and irrational use of antibiotics in human veterinary medicine has seriously endangered the ecological environment and human health.In this study,a fully automatic solid-phase disk extraction-stable isotope dilution-ultra-high performance liquid chromatography-mass spectrometry method for determination of 17 kinds of sulfonamides antibiotics(SAs)in water was established,which was then applied to determination of SAs in real samples including tap water,river water and seawater,respectively.Meanwhile,the residual characteristics were discussed and the ecological risks were assessed.With this method,1.0 L water sample with 0.5 g/L Na2EDTA(pH=3)was extracted and enriched by 3M SDB-XC disk,and eluted by 10 mL of mixture of methanol and acetone(1:1,V/V),and the pretreatment time of the sample was about 60 min per six samples.Under the optimized conditions,the linearity of the method for detection of 17 kinds of SAs ranged from 0.05 to 100 μg/L,with correlation coefficients(R2)>0.999.In addition,the detection limits(S/N=3)were as low as 0.012-0.052 ng/L,and the recoveries were in the range of 76%-110%,with relative standard deviations of 0.5%-9.6%(n=5).The results showed that no SAs was detected in tap water,while 3 and 9 kinds of SAs were detected in river water of Zhoushan,and seawater of Wenzhou Sea area in Zhejiang province,respectively.The total concentrations of the detected SAs were 0.875-21.826 ng/L and 1.024-20.768 ng/L in river water and seawater,respectively,and among which,sulfamethoxazole(SMX)was the predominant compound in river water and seawater,accounting for 81%and 74%of the total SAs,respectively.The ecological risk assessment showed that the risk quotients of the detected SAs in the river water and seawater in the study area for the three kinds of trophic organisms(algae,Daphnia and fish)were far less than 0.01,meaning that the ecological risk was low.

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires

This study investigated the effects of different frequencies of repetitive transcranial magnetic stimulation (rTMS) on chronic neuropathic pain in rats.The behavior of rats with experimental chronic neuropathic pain was observed,and the expression of neuronal nitric oxide synthase (nNOS) in the ipsilateral dorsal root ganglions (DRGs) and the activation and proliferation of astrocytes in the ipsilateral spinal dorsal horn were detected.Thirty-two male Sprague-Dawley rats were randomly divided into four groups:sham-operated group,sham-rTMS group,1 Hz group and 20 Hz group (8 rats in each group).Chronic constriction nerve injury induced by sciatic nerve ligation was made to establish the models of the chronic neuropathic pain in rats except those in the sham-operated group.Then we applied different frequencies of rTMS to the primary motor cortex (M1) contralateral to the pain side once daily for 10 consecutive days.Pain behavior scores were observed before and after treatment.Western blot analysis was used to detect the expression of nNOS in ipsilateral L4-6 DRGs.Double immunofluorescent labeling for glial fibrillary acidic protein (GFAP) and 5-bromo-2-deoxyuridine (BrdU) was employed to observe the activation and proliferation of astrocytes in the ipsilateral L4-6 spinal dorsal horn.After rTMS treatment,the spontaneous pain behavior scores were significantly lower in the 20 Hz group than those in the sham-rTMS group (P＜0.05).Moreover,the brush-evoked pain behavior scores were significantly lower in the 20 Hz group than those in the sham-rTMS and 1 Hz group (P＜0.05),suggesting that the spontaneous pain and brush-evoked pain in the 20 Hz group were significantly alleviated.Western blot analysis revealed that the expression of nNOS in ipsilateral L4-6 DRGs was significantly decreased in the 20 Hz group as compared with the sham-rTMS group and the 1 Hz group (P＜0.01) after rTMS treatment.Double immunofluorescence suggested that the expression of GFAP and the co-localization with BrdU in astrocytes were less in the sham-operated group than those in the sham-rTMS group and the 1 Hz group in L4-6 spinal dorsal horn ipsilateral to the neuropathic pain.After rTMS treatment,the expression of GFAP and the co-localization with BrdU decreased in the 20 Hz group as compared with the sham-rTMS group and the 1 Hz group (P＜0.05).In addition,the alleviation degree of spontaneous pain and brush-evoked pain in the 20 Hz group was negatively correlated with the expression of nNOS in ipsilateral DRGs and the number of GFAP/BrdU co-labelled astrocytes in L4-6 spinal dorsal horn ipsilateral to the neuropathic pain (P＜0.05).It was suggested that high-frequency rTMS may relieve neuropathic pain through down-regulating the overexpression of nNOS in ipsilateral DRGs and inhibiting the activity and proliferation of astrocytes in L4-6 spinal dorsal horn ipsilateral to the neuropathic pain.

Salviae Miltiorrhizae Radix et Rhizoma (Danshen) is one of the commonly used bulk medicinal materials in China. It is widely used in clinical practice, and has many pharmacological effects, such as antithrombotic，antibacterial，anti-inflammatory，anti-oxidative and immunomodulatory activities. The quality of Danshen determines the quality of the curative effect. The current wild resources of Danshen gradually decrease, the cultivation area is widely distributed, among them, Shandong, Sichuan, Anhui, Shanxi, Hebei, Henan and other provinces have large acreage for Danshen. However, germplasm, origin, cultivation, harvest, process and other factors have a certain impact on the active ingredients of Danshen, which lead to the quality of Danshen good and bad mixed. This paper is based on the systematic analysis of literature on quality evaluation of Danshen. To provide a new idea for the further research on the quality of Danshen, we summarized the main factors affecting the quality of Salvia miltiorrhiza that of germplasm, origin, cultivation, harvest, process, storage, product specifications and so on.

Epimedii Folium has a long history in China as a common traditional Chinese medicine. Key factors of Epimedii Folium quality were summarized based on ancient literatures, Chinese Pharmacopoeias and modern research in different period of history. The main reason for unqualified Epimedii Folium is unstable icariin. Therefore, it's suggested that: the precondition of the quality control of epimedium is to find the proper quality marker. It's suggested that the medicinal parts should be reverted to "dry whole plant overground" to solve Epimedium resource shortage problem. In addition, it is necessary to strengthen the standardized cultivation, so as to ensure germplasm, production area, and producing method to guarantee the quality of Epimedium Folium. In the drying method, it is recommended to change "dry in the sun or shade" to "dry", namely dry in the sun, shade or drier, in order to provide a new method to improve the quality control and quality standard of Epimedii Folium.

OBJECTIVETo explore the features of immunophenotypes and the characteristics of molecular biology and cellular genetics of AML patients with CD7 and CD4 expression.

METHODSThe immunophenotypical markers of AML cells were detected by multiple parameter flow cytometry; the expression of WT1, MDK, ETO, PML-RaRa and BCR-ABL were detected by RT-PCR; and cellular features were analyzed by R-band in 304 patients. The patients were divided into three groups according to their immunophenotypes: AML with CD7 expression (CD7 group), AML with CD4 expression(CD4 group) and AML without CD7 and CD4 expression (common AML group).

RESULTSThe expression rate and level of HLA-DR in CD7 group were higher than those in the common AML group, and the expression rate of CD33 and CD34 was higher than that in the other two groups. The expression rate and level of CD15, CD64 in the CD4 group were higher than those in the other 2 groups, and the expression rate and level of CD33 were higher than those in the common AML group. WT1 expression in the CD7 group was lower than that in the common AML group. PML-RaRa was not detected in the CD7 group. AML with co-expression of CD4 or CD7 showed more normal karyotype. (15;17) was not found in AML with CD7 expression.

CONCLUSIONAML cells with CD7 expression originate from precursor cells and are blocked in the early phase of hematological development; AML cells with CD4 expression originate from more mature stage of hematological devevelopment and with CD33, CD64 and CD15 high expression; AML cells with CD7 and CD4 expression are characterized by no-specific change of cellular genetics. According to the expression level and intesity of CD4 and CD7, and together with other specific lineage markers, the MRD in AML patients can be quantitatively detected.

OBJECTIVETo investigate the effect of premature rupture of the membrane (PROM) on neonatal complications in premature infants.

METHODSThe registration information of 7684 preterm infants with gestational age <37 weeks were collected from the cooperative units in the task group between January 1, 2014 to December 31, 2014. Specially trained personnel from each cooperative units filled in the unified form in a standardized format to record the gender, gestational age, birth weight, PROM, placental abruption, antenatal corticosteroid, Apgar score, amniotic fluid pollution, and complications of the infants. The data were analyzed comparatively between the cases with PROM and those without (control).

RESULTSThe preterm mortality rate was significantly lower but the incidences of ICH, NEC, ROP and BPD were significantly higher in PROM group than in the control group (P<0.05). The 95% confidence interval of the OR value was <1 for mortality, and was >1 for ICH, NEC, ROP and BPD. After adjustment for gestational age, birth weight, gender, mode of delivery, placental abruption, placenta previa, prenatal hormones, gestational diabetes mellitus (GDM), gestational period hypertension and 5-min Apgar score <7, the incidences of NEC, ROP and BPD were significantly different between the two groups (P<0.05) with 95% confidence interval of OR value >1, but the mortality rate and incidence of ICH were not significantly different between the two groups (P>0.05).

CONCLUSIONPROM is a risk factor for NEC, ROP and BPD in preterm infants, and adequate intervention of PROM can reduce the incidences of such complications as NEC, ROP and BPD in the infants.

Apgar Score ; Birth Weight ; Female ; Fetal Membranes, Premature Rupture ; pathology ; Gestational Age ; Humans ; Incidence ; Infant, Newborn ; Infant, Newborn, Diseases ; etiology ; Infant, Premature ; Pregnancy ; Risk Factors

BACKGROUNDAlcohol dependence (AD) is a complex disorder characterized by impaired control over drinking. It is determined by both genetic and environmental factors. The recent approach of genome-wide association study (GWAS) is a powerful tool for identifying complex disease-associated susceptibility alleles, however, a few GWASs have been conducted for AD, and their results are largely inconsistent. The present study aimed to screen the loci associated with alcohol-related phenotypes using GWAS technology.

METHODSA genome-wide association study with the behavior of regular alcohol drinking and alcohol consumption was performed to identify susceptibility genes associated with AD, using the Affymetrix 500K SNP array in an initial sample consisting of 904 unrelated Caucasian subjects. Then, the initial results in GWAS were replicated in three independent samples: 1972 Caucasians in 593 nuclear families, 761 unrelated Caucasian subjects, and 2955 unrelated Chinese Hans.

RESULTSSeveral genes were associated with the alcohol-related phenotypes at the genome-wide significance level, with the ankyrin repeat domain 7 gene (ANKRD7) showing the strongest statistical evidence for regular alcohol drinking and suggestive statistical evidence for alcohol consumption. In addition, certain haplotypes within the ANKRD7 and cytokine-like1 (CYTL1) genes were significantly associated with regular drinking behavior, such as one ANKRD7 block composed of the SNPs rs6466686-rs4295599-rs12531086 (P = 6.51 × 10(-8)). The association of alcohol consumption was successfully replicated with rs4295599 in ANKRD7 gene in independent Caucasian nuclear families and independent unrelated Chinese Hans, and with rs16836497 in CYTL1 gene in independent unrelated Caucasians. Meta-analyses based on both the GWAS and replication samples further supported the observed significant associations between the ANKRD7 or CYTL1 gene and alcohol consumption.

CONCLUSIONThe evidence suggests that ANKRD7 and CYTL1 genes may play an important role in the variance in AD risk.

Adult ; Aged ; Alcohol Drinking ; genetics ; Blood Proteins ; Cytokines ; Female ; Genome-Wide Association Study ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Proteins ; genetics ; Receptors, Cytokine ; genetics

S-phase kinase-associated protein 2 (Skp2), which plays a role in cell cycle regulation, is commonly overexpressed in a variety of human cancers and associated with poor prognosis. However, its role in nasopharyngeal carcinoma (NPC) is not well understood. In this study, we examined the clinical significance of Skp2, with a particular emphasis on overall survival (OS) and disease-free survival (DFS), in NPC cases in South China, where NPC is an epidemic. Additionally, we explored the function of Skp2 in maintaining a cancer stem cell-like phenotype in NPC cell lines. Skp2 expression was assessed for 127 NPC patients using tissue microarrays and immunohistochemistry and analyzed together with clinicopathologic features, OS, and DFS. Skp2 expression was detectable, or positive, in 75.6% of patients. Although there was no correlation between Skp2 and any clinicopathologic factor, Skp2 expression significantly portended inferior OS (P = 0.013) and DFS (P = 0.012). In the multivariate model, Skp2 expression remained significantly predictive of poor OS [P = 0.009, risk ratio (RR) = 4.06] and DFS (P = 0.008, RR = 3.56), and this was also true for clinical stage (P = 0.012 and RR=3.201 for OS; P = 0.002 and RR=1.94 for DFS) and sex (P = 0.016 and RR=0.31 for OS; P = 0.006 and RR = 0.27 for DFS). After Skp2 knockdown, a colony formation assay was used to evaluate the self-renewal property of stem-like cells in the NPC cell lines CNE-1 and CNE-2. The colony formation efficiency in CNE-1 and CNE-2 cells was decreased. In Skp2-transfected CNE-1 and CNE-2 cells, side population (SP) proportion was increased as detected by flow cytometry. Skp2 is an independent prognostic marker for OS and DFS in NPC. Skp2 may play a role in maintaining the cancer stem cell-like phenotype of NPC cell lines.
Adolescent ; Adult ; Aged ; Carcinoma ; Cell Line, Tumor ; China ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gene Knockdown Techniques ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Neoplasm Staging ; Neoplastic Stem Cells ; pathology ; RNA, Small Interfering ; genetics ; S-Phase Kinase-Associated Proteins ; genetics ; metabolism ; Sex Factors ; Survival Rate ; Tissue Array Analysis ; Transfection ; Young Adult

OBJECTIVETo observe the mechanical properties of the prefabricated connective tissue tube as blood vessel substitute and its changes after implantation at the femoral artery.

METHODSThe acellular matrix tube of 8-12 cm in length with a silicone rod inside it was implanted into dog peritoneal cavity. 3 weeks later, a new formed tube around the silicone rod was transferred to the femoral artery as blood vessel substitute. The mechanical properties and histological examination of the blood vessel substitute were assessed and compared to those of the carotid artery and vein. 6 months after transfer, the patency of the blood vessels substitute was observed. The histological change was studied by light microscopy, scanning and transmitting electron microscopy.

RESULTS(1) The mechanical properties of blood vessel substitute was not as strong as artery, but better than the vein. (2) There were elastic and collagen fibers with many fibroblasts around the tube wall, but few mesothelial cells around the inner wall. All of the blood vessel substitutes (n = 6) were found to keep patency and the structure of the blood vessels substitutes became similar to femoral artery 6 months after they had been grafted to the femoral artery.

CONCLUSIONSThese results suggest that tissue engineering in vivo is a good approach to construct vessels substitute. The tissue tubes made in dog's peritoneal cavity have good condition when it is used as a blood vessel substitutes.

Animals ; Blood Vessel Prosthesis ; Blood Vessels ; transplantation ; Carotid Arteries ; surgery ; Dogs ; Extracellular Matrix ; Tissue Engineering ; methods