Adult ; Cross-Sectional Studies ; Female ; Humans ; Job Satisfaction ; Nurses ; Ophthalmology ; manpower ; Stress, Psychological ; Surveys and Questionnaires ; Workload ; Young Adult

OBJECTIVETo investigate the effect of Xuezhikang (, XZK) on renal cell apoptosis in diabetic rats and the possible mechanism.

METHODSSixty-six rats were randomly divided into 3 groups: the normal, model and XZK groups. In each group, the rats were further randomly divided into 3-month and 6-month subgroups, respectively. Diabetes of rats was induced by a single intraperitoneal injection of 1% streptozocin at 60 mg/kg body weight. Rats in the XZK group received gastric perfusion of XZK (1200 mg/kg body weight) everyday for 3 or 6 months, while rats in the normal and model groups received equal volume of saline. Twenty-four hours' urine was collected for urinary albumin excretion (UAE) measurement. Periodic acid-Schiff (PAS) and Masson's trichrome staining were used for saccharides and collagen detection. Cell apoptosis of renal cortex was investigated by TdT-mediated dUTP nick end labeling (TUNEL) staining. Bax and Bcl-2 expressions were detected by immunohistochemistry and Western blot, respectively. Cytochrome C (Cyt C) and caspase-9 concentration were detected by Western blot.

RESULTSCompared with the model group, XZK treatment could significantly decrease the kidney hypertrophy index, 24 h UAE, renal cell apoptosis, cytoplasmic Cyt C level and active caspase-9 level, as well as suppress the increment of Bax and up-regulate the expression of Bcl-2, leading to the suppression of Bax/Bcl-2 ratio at 3 and 6 months (P<0.05 or P<0.01). Moreover, XZK treatment could alleviate the deposition of PAS-stained saccharides and Masson's trichromestained collagen to different extent.

CONCLUSIONSRenal cell apoptosis was observed in diabetic kidney, in which mitochondrial apoptotic pathway might be involved. XZK treatment could attenuate pathological changes in diabetic kidney and reduce renal cell apoptosis, probably via the suppression of Bax/Bcl-2 ratio, which lead to inhibition of Cyt C release and following caspase-9 activation.

Albuminuria ; blood ; complications ; Animals ; Apoptosis ; drug effects ; Blood Glucose ; metabolism ; Caspase 9 ; metabolism ; Cytochromes c ; metabolism ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hypertrophy ; In Situ Nick-End Labeling ; Kidney ; drug effects ; pathology ; Kidney Glomerulus ; pathology ; Lipids ; blood ; Male ; Mesangial Cells ; drug effects ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats, Sprague-Dawley ; Streptozocin ; bcl-2-Associated X Protein ; metabolism

Common bile duct (CBD) stones are a frequent problem in Chinese populations, and their incidence is particularly high in certain areas (Wang et al., 2013). In recent years, laparoscopic common bile duct exploration (LCBDE) and endoscopic retrograde cholangiopancreatography (ERCP) have been the main surgical procedures for CBD stones, although each has different advantages and disadvantages in the treatment of choledocholithiasis (Loor et al., 2017; Zhou et al., 2017). For patients with large stones, a dilated CBD, especially concurrent gallstones, LCBDE is the preferred and most economical minimally invasive procedure (Koc et al., 2013). However, a T-tube is often placed during LCBDE to prevent postoperative bile leakage; this is associated with problems such as bile loss, electrolyte disturbance, and decreased gastric intake (Martin et al., 1998). In addition, the T-tube usually must remain in place for more than a month, during which time the patient's quality of life is seriously compromised. Many skilled surgeons currently perform primary closure of the CBD following LCBDE, which effectively speeds up rehabilitation (Hua et al., 2015). However, even in sophisticated medical centers, the incidence of postoperative bile leakage still reaches ≥10% (Liu et al., 2017). Especially for a beginner, bile leakage remains a key problem (Kemp Bohan et al., 2017). Therefore, a safe and effective minimally invasive surgical approach to preventing bile leakage during primary closure of the CBD after LCBDE is still urgently needed.
Aged ; Aged, 80 and over ; Choledocholithiasis ; Common Bile Duct Diseases ; Drainage/methods* ; Female ; Gallstones ; Gastroscopy ; Humans ; Laparoscopy ; Male ; Middle Aged

OBJECTIVETo construct prokaryotic expression systems of Streptococcus pneumoniae ciaH and ciaR genes,and to determine their correlation with drug resistance.

METHODSThe total length of ciaH and ciaR genes was amplified by PCR and their prokaryotic expression systems were established by using routine genetic engineering technique. SDS-PAGE was applied to measure the outputs of target recombinant proteins rCiaH and rCiaR. Rabbits antisera and IgGs against rCiaH and rCiaR were prepared. The resistance to penicillin and cefotaxime of S.pneumoniae strains was examined after CiaH and CiaR were extracellularly and intracellularly blocked by the IgGs.

RESULTThe homogeneity of nucleotide and amino acid sequences of the cloned ciaH and ciaR genes with the reported sequences was 99.9-100% and 100%, respectively. The recombinant bacteria E.coli BL21DE3pET42a-ciaH and E.coli BL21DE3pET42a-ciaR were able to express the target recombinant proteins rCiaH and rCiaR with efficiency. The outputs of rCiaH and rCiaR were 33% and 45% of the total bacterial proteins, respectively. The double immunodiffusion titers of rCiaH antiserum,rCiaR antiserum,rCiaH-IgG and rCiaR-IgG were 1:4,1:4,1:1 and 1:1, respectively. After CiaH was extracellularly or intracellularly blocked by CiaH-IgG, and CiaR was intracellularly blocked by CiaR-IgG, the penicillin-sensitive or cefotaxime-sensitive strains developed resistance to the two antibiotics; but the blocks did not change that of penicillin-resisting or cefotaxime-resisting strains.

CONCLUSIONThe prokaryotic expression systems of S. pneumoniae ciaH/ciaR genes have been successfully constructed in this study. Both the CiaH and CiaR may be involved in penicillin and cefotaxime resistance of the bacterium.

Animals ; Bacterial Proteins ; genetics ; metabolism ; Cefotaxime ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Genetic Vectors ; Penicillin Resistance ; genetics ; Protein Kinases ; genetics ; metabolism ; Rabbits ; Recombinant Fusion Proteins ; genetics ; metabolism ; Signal Transduction ; Streptococcus pneumoniae ; drug effects ; enzymology ; genetics ; beta-Lactam Resistance ; genetics

OBJECTIVECentral obesity is considered to be a central component of metabolic syndrome. Waist circumference (WC) has been widely used as a simple indicator of central obesity. This study is aimed to evaluate the sensitivity of WC cut-off values for predicting metabolic risk factors in middle-aged Chinese.

METHODSThe study involved 923 subjects aged 40-65 years. The metabolic risk factors were defined according to the Chinese Joint Committee for Developing Chinese Guidelines on Prevention and Treatment of Dyslipidemia in Adults. WC cut-off 85-90 cm and ⋝90 cm were used as cut-off values of central pre-obesity and central obesity in males, respectively, while WC 80-85 cm and ⋝85 cm were used as cut-off values of central pre-obesity and central obesity in females.

RESULTSFirst, WC values corresponding to body mass index (BMI) 24 kg/m2 and visceral fat area (VFA) 80 cm2 were 88.55 cm and 88.51 cm in males, and 81.46 cm and 82.51 cm in females respectively. Second, receiver operating characteristic curves showed that the optimal WC cut-off of value was 88.75 cm in males, higher than that in females (81.75 cm). Third, the subjects with higher WC values were more likely to have accumulating metabolic risk factors. The prevalence of metabolic risk factors increased linearly and significantly in relation to WC levels.

CONCLUSIONWC cut-off values of central pre-/central obesity are optimal to predict multiple metabolic risk factors.

Aged ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Intra-Abdominal Fat ; physiopathology ; Male ; Metabolic Syndrome ; diagnosis ; epidemiology ; physiopathology ; Middle Aged ; Obesity ; diagnosis ; ROC Curve ; Waist Circumference

Objective To investigate the different characteristics of apoptosis during the different phases of cell cycle between astrocytes and neurons, so as to explore the apoptosis mechanism of the 2 kinds of cells. Methods Ischemia/reperfusion injury was induced by temporary middle cerebral artery occlusion (MCAO). And the brain was taken 3 d after injury. Thymidine analogues such as bromodeoxyuridine (BrdU) was used to incorporate during the S phase of the cell cycle, and the combined visualization of BrdU labeling and Tdt-mediated dUTP-biotin nick end labeling was performed to investigate the apoptosis in the adult rat brain in vivo. Then the apoptosis during the different phases of cell cycle between astrocytes and neurons was detected by flow cytometry (FCM) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods. Results Compared to the normal controls, BrdU or TUNEL positive cells were increased in injury group, and a few of of BrdU and TUNEL double positive cells were found. The most apoptotic neurons and astrocytes were in G0/G1 phase, and the apoptotic neurons were less than astrocytes in G0/G1 phase (P＜0.01), but the apoptotic neurons were more than astrocytes in S phase (P＜0.01). Conclusions Astrocytes and neurons may have the different characteristics of apoptosis during the different phases of cell cycle, which is induced at different check-points.

Objective To explore the mechanism of opioid-psychic dependence involving the aspects of pre-receptor and receptor by observing the changeable expressions of glutamate neurotransmitter and NR2B of N-methyl-D-aspartic acid (NMDA) receptor in the neuroanatomical circuit of ventral temental area, nucleus accumbens and prefrontal cortex (VTA-Nac-PFC) of rats subjected to morphine-induced conditioned place preference (CPP). Methods The models of CPP were validated by escalating doses of morphine in rats (n=16). The colorimetry and immunohistochemistry ways were applied to detect the contents of glutamic acid and the expression level of NR2B in VTA, Nac and PFC. Results As compared with those in the control group physiological saline), the prolonged detention time of white compartment in the model group was notably observed (P<0.05), and increased content of glutamic acid and expression level of NR2B in fields of VTA, Nac and PFC in the model group were significantly detected (P<0.05). Conclusion Increased level of giutamic acid and expression level of NR2B in nuroanatomieal circuit of VTA, Nac and PFC could play key roles in inducing morphine-psychic dependent rats.

Pancreatic cancer remains one of the most lethal malignancies worldwide. The combination of the first-line standard agent gemcitabine (GEM) with the molecular-targeted drug erlotinib (Er) has emerged as a promising strategy for pancreatic cancer treatment. However, the clinical benefit from this combination is still far from satisfactory due to the unfavorable drug antagonism and the fibrotic tumor microenvironment. Herein, we propose a membrane-camouflaged dual stimuli-responsive delivery system for the co-delivery of GEM and Er into pancreatic cancer cells and tissues to block the antagonism, as well as reshapes profibrotic tumor microenvironment via simultaneous delivery of small interference RNA (siRNA) for synergistic pancreatic cancer treatment. This "all-in-one" delivery system exhibits sensitive GSH and pH-dependent drug release profiles and enhances the inhibitory effects on the proliferation and migration of tumor cells in vitro. Excitingly, the systemic injection of such a biomimetic drug co-delivery system not only resulted in superior inhibitory effects against orthotopic pancreatic tumor and patient-derived tumor (PDX), but also greatly extended the survival rate of tumor-bearing mice. Our findings provide a promising therapeutic strategy against pancreatic cancer through the enhanced synergistic effect of target therapy, chemotherapy and anti-fibrotic therapy, which represents an appealing way for pancreatic cancer treatment.

OBJECTIVETo establish a 16S rDNA multiplex polymerase chain reaction (PCR) for simultaneously detecting P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens of chronic periodontitis and to study the correlation between different modes of infection and severity of the disease.

METHODSPeriodontal pocket specimens from 152 patients with mild, moderate or advanced chronic periodontitis and gingival sulcus specimens from 30 periodontally healthy individuals were collected and placed in 200 microl lysis buffer. The specimens were incubated in 100 degrees C for 10 min and 10 microl of the supernatant was directly used as PCR template. DNAs from P. gingivalis strain ATCC33277, A. actinomycetemcomitans strain Y4, T. denticola strain FM and E. coli strain DH5alpha were used as positive and negative controls in PCR with all of which were prepared by routing phenol-chloroform method. A multiplex PCR assay, using three sets of primers specific to 16S rDNA genes of the three anaerobes, was developed to detect the specimens. The target amplification fragments from 3 cases of PCR products positive for all the three anaerobes were sequenced after T-A cloning. Chi-square test was applied to analyze the correlation between different coinfection of the three anaerobes and severity of the disease.

RESULTSThe established 16S rDNA multiplex PCR assay was able to detect P. gingivalis, A. actinomycetemcomitans and T. denticola at a minimum of 10, 20 and 20 cells, respectively. In comparison with the reported corresponding sequences, similarities of the nucleotide sequences from the three anaerobes 16S rDNA amplification fragments were as high as 99.45%, 97.08% and 96.59%, respectively. Of the 30 periodontally healthy gingival sulcus specimens, only one (3.3%) positive for P. gingivalis and two (6.7%) for A. actinomycetemcomitans could be identified but all of the rest were negative. In the 152 CP periodontal pocket specimens, 147 cases (96.7%) were positive for P. gingivalis, A. actinomycetemcomitans and/or T. denticola, respectively, and 5 cases (3.3%) were negative for all the three anaerobes. The positive rate of P. gingivalis detection (91.5%, 139/152) was significantly higher than those of A. actinomycetemcomitans (72.4%, 110/152) and T. denticola (80.9%, 123/152) (chi(2) = 7.07, 18.67; P < 0.01). 89.8% of the specimens from patients showed coinfections with two (26.5%) or three anaerobes (63.3%), and the coinfection rates in the specimens from moderate and advanced CP were remarkably higher than that from mild CP (chi(2) = 10.43, P < 0.01).

CONCLUSIONThe 16S rDNA multiplex PCR established in this study showed high sensitivity and specificity, which could be used to detect P. gingivalis, A. actinomycetemcomitans and T. denticola in clinical specimens. CP was an disease caused by multiple pathogenic microbes while the synergistic pathopoiesis of the three microbes was closely related to the severity of the disease.

Actinobacillus Infections ; epidemiology ; microbiology ; Adult ; Aged ; Aggregatibacter actinomycetemcomitans ; isolation & purification ; Bacteroidaceae Infections ; epidemiology ; microbiology ; China ; epidemiology ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Periodontitis ; microbiology ; Polymerase Chain Reaction ; methods ; Porphyromonas gingivalis ; isolation & purification ; RNA, Ribosomal, 16S ; Treponema denticola ; isolation & purification ; Treponemal Infections ; microbiology

Background Retinitis pigmentosa (RP) is a monogenic inheritance and blinding disease of fundus oculi.There is not an effective therapeutic method now.Objective This work was to identify the mutations of RP1 gene in Chinese RP patients in Ningxia area and to explore the potential interactions in the pathogenesis of RP.Methods The periphery blood of 3-5 ml was collected from 110 individuals with RP(35 ADRP and 75SRP)and 100 normal controls in Ningxia area.Polymerase chain reaction (PCR) and direct DNA sequencing were used to screening the sequence alterations in the entire coding region and splice sites of RP1 gene.Multivariate analysis and two web-based programs( PolyPhen and SIFT) were used to analyze the results.Results Eleven mutation locus were detected in the exon 4 of RP1 gene including two novel sequence variants:p.Lys1152Lys without a higher mutation rate in comparison with normal control group(x2 =9.12 P＜0.01 ),but c.* 247A＞C with a higher mutation rate in comparison with normal control group(x2 =12.77,P＜0.01 ) and c.* 247A＞C mutation was thought to be correlated with RP( r=1.11,P＜0.05 ).The other ten mutation locus were reported as single nucleotide polymorphisms (SNP).The mutation rate of p.Gln1725Gln was found to be higher in the RP patients than the normal controls (x2 =42.09,P＜0.01 ),but no the significant correlation was seen between the pathogenesis of RP and mutation of p.Gln1725Gln(r=1.74,P＞0.05).p.Lys1152Lys mutation was found in only 1 patient.Three SNPs( p.Arg872His,Ala1670Thr,Ser1691Pro) were always occurred in the same 83 RP patient and the relevance ratio was higher than controls ( P＜0.01 ).The age of night blindness on patients with concurrent three mutations was (30.54± 13.68 ) years,and the best corrected visual acuity (BCVA) was 0.50 ± 0.38.The age of night blindness on patients without concurrent three mutations was(21.06± 16.24) years,and the BCVA was 0.40 ±0.33 and were higher than controls ( t =2.11,P ＜ 0.05 ).Conclusions In this study,the prevalence of RP1 mutations among the RP patients in Ningxia population was lower than other populations (＜ 1％ ).The alliance of SNPs (p.Arg872His、p.Ala1670Thr、p.Ser1691Pro) may play a protective role on RP patients and reduce the frequency of mutatiaon in RP1 gene.