Objective To study the construction of the lentiviral-mediated RNA interference(RNAi) vector targering rat suppressors of cytokine signaling 3(SOCS3) gene.Methods Three target sequences were selected by on-line designer software on Ambion according to rat SOCS3 mRNA sequence(NM053565),the complementary DNA contained both sense and antisense oligonucleotides were designed and synthesized.After annealing,these double strands DNA were cloned to pRNA-Lenti-green fluorescent protein(GFP),which contained U6 promoter and GFP.The resulting Lentiviral vector containing SOCS3 shRNA was named pRNA-Lenti-SOCS3-GFP.After the rat glioma cells(C6)were transduced with the constructed 1entiviral vectors,real-time polymerase chain reaction was used to evaluate the level of SOCS3 expression(including siRNA1 group,siRNA2 group,siRNA3 group,vacuity group and siRNA-Negative group).The pRNA-Lenti-SOCS3-GFP and Lentivector Pakaging plasmid mix were cotransfected into 293T to package Lentivirus particles.Culture supematant was harvested,then the virus titer was determined by serial dilution assay.Results The SOCS3 mRNA sequence was successfully cloned to pRNA-Lenti-GFP,which was proved by PCR and DNA sequence.Compared with control group,the SOCS3 mRNA expressions were obviously suppressed in all 3 experimental groups,especially the expression rate in siRNA1 group was reduced by 80%.The Lentiviral particle titer was determined by serial dilution assay with 1.0?1010 TU?L-1.Conclusion The lentiviral-mediated RNAi vector of rat SOCS3 gene has been constructed successfully,this may provide a potential tool for studying and treating SOCS3-related diseases.

Objective To evaluate the relationship between postoperative cognitive dysfunction and inflammatory responses of patients with non-acute fragile brain function. Methods Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients of both sexes, aged 65-83 yr, weighing 52-85 kg, scheduled for elective lumbar spinal internal fixation under general anesthesia, were divided into 2 groups (n=30 each) using a random number table method: brain function normal control group (group C) and non-acute fragile brain function group ( group F) . The blood samples were obtained from the peripheral vein at 30 min before anesthesia induction, at the end of operation and at 2 days after operation to determine the concentrations of tumor necrosis factor-alpha ( TNF-α) , interleukin-6 ( IL-6) , IL-10 and C-reactive pro-tein by enzyme-linked immunosorbent assay. Confusion assessment method was used to assess the develop-ment of delirium at 2 days after operation. Mini Mental State Examination was used to assess the develop-ment of postoperative cognitive dysfunction at 7 days after operation. Results Compared with the baseline at 30 min before anesthesia induction, the concentrations of TNF-α, IL-6 and C-reactive protein in plasma were significantly increased at 2 days after operation in two groups ( P<0. 05) . Compared with group C, the concentrations of TNF-α and IL-6 in plasma and incidence of postoperative delirium were significantly in-creased at 2 days after operation in group F ( P<0. 05) . Conclusion The mechanism of delirium occurred after surgery may be related to inflammatory responses of patients with non-acute fragile brain function.

OBJECTIVESTo screen the hALR-interacting protein by phage-displayed technique and identify its biological activities.

METHODSThe specific phage clones that interacted with target protein hALR from a cDNA library of hepatocarcinoma cells were selected using the T7 phage-displayed technique. The acquired cDNA inserts were sequenced and analyzed by bioinformatic tools. The biological activities of the phage-displayed peptide affecting QGY hepatocarcinoma cells were studied using 3H-TdR method.

RESULTSThe cDNA inserts with 212 bp were acquired after 4 rounds of biopanning. They showed 100% homology with citron kinase. The phage-displayed peptide and the peptide combined with hALR affected QGY cells proliferation.

CONCLUSIONShALR-interacting peptide can be specifically screened by phage-displayed technique. Citron kinase that interacted with hALR potentially plays an important role in the proliferation of hepatocarcinoma cells.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Gene Library ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Liver Regeneration ; Peptide Library ; Peptides ; metabolism ; Protein Binding ; Proteins ; genetics ; isolation & purification ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo investigate the dynamic changes of a-AR, b1-AR and b2-AR expression in hepatic fibrosis.

METHODSRat hepatic fibrosis model was established by bile duct ligation (BDL). HE and Masson staining were used to determine hepatic fibrosis levels. Immunohistochemistry was applied to detect alpha -smooth muscle actin (alpha -SMA), a marker of hepatic stellate cell (HSC) activation; Western blot and real-time RT-PCR were used to measure the dynamic changes of alpha -AR, beta(1)-AR, beta(2)-AR expression on protein and mRNA levels, respectively, during the development of hepatic fibrosis.

RESULTS(1) HE and Masson trichrome staining showed that the liver fibrosis models were established successfully. (2) At 1, 2, 3, 4 wk after BDL, alpha -SMA positive area density of the model group (10.58% +/- 1.75%, 24.14% +/- 2.02%, 29.74% +/- 2.59%, 34.28% +/- 2.01%) was significantly higher than that of the sham operation group (4.12% +/- 1.51%), P less than 0.01. (3) The expression of alpha -AR, beta(1)-AR, beta(2)-AR protein and mRNA was increased with the development of the hepatic fibrosis (P less than 0.05). (4) alpha -SMA expression was positively associated with alpha -AR, beta(1)-AR, beta(2)-AR, r values were 0.564, 0.753 and 0.606, respectively.

CONCLUSIONThe expression of alpha -SMA is increased dramatically during the fibrosis, and is positively associated with the expression of alpha -AR, beta(1)-AR and beta(2)-AR.

Actins ; metabolism ; Animals ; Hepatic Stellate Cells ; metabolism ; pathology ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Biliary ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Polymerase Chain Reaction ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, alpha ; genetics ; metabolism ; Receptors, Adrenergic, beta ; genetics ; metabolism ; Sympathetic Nervous System ; metabolism ; Time Factors

OBJECTIVETo evaluate the effects of fluorid on morphology change in enamel and dentin during tooth bleaching.

METHODSThe study population consisted of twelve patients who required the extraction of first premolars for orthodontic reasons. Twelve participants were divided into three groups: bleaching with NaF-treated group, bleaching-treated group and control group. Immediately after bleaching treatment, all teeth were extracted and prepared for scanning electron microscope (SEM). Morphologic observations were carried out with SEM.

RESULTSIn the bleaching-treated group, mild demineralization was observed on the surface of enamel and collapse of collagen scaffold was also observed on the longitudinal section of dentine. The diameter of dentinal tubule was not uniform due to peritubular dentine was demineralized. In the bleaching with NaF-treated group, the demineralization of enamel and dentin were reduced and some diameter of dentinal tubule were smaller than bleaching-treated group.

CONCLUSIONFluoride can reduced the demineralization of enamel and dentine obviously, which may be applied as a therapeutic tool for sensitivity induced by tooth bleaching.

Dental Enamel ; Dentin ; Fluorides ; Humans ; Microscopy, Electron, Scanning ; Phosphates ; Tooth Bleaching

Objective:To evaluate the effect of general anesthesia guided by electroencephalography (EEG) monitoring on postoperative delirium (POD) in elderly patients with non-acute fragile brain function.Methods:Sixty patients of both sexes with non-acute fragile brain function, aged 65-85 yr, of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with body mass index of 18-30 kg/m 2, undergoing hip replacement, were divided into 2 groups ( n=30 each) by a random number table method: conventional general anesthesia group (group C) and general anesthesia guided by EEG monitoring group (group E). Anesthesia was maintained by intravenous infusion of propofol 50-150 μg·kg -1·min -1 and remifentanil 0.05-0.30 μg·kg -1·min -1 and intermittent intravenous boluses of rocuronium.In group E, the dose of anesthetic was reduced when the EEG burst-suppression ratio≥10% for more than 1 min or anesthesia index (AI) <40.If the situation mentioned above still existed after 1 min, the dose of anesthetic was continued to be reduced or norepinephrine was injected intravenously.In group C, the amount of intraoperative anesthesia was adjusted according to the changes in hemodynamics.Norepinephrine 4-10 μg or dopamine 1 mg was given intravenously in the light of the patients′ heart rates when intraoperative hypotension occurred.At 10 min after anesthesia induction, immediately after skin incision, immediately at the end of surgery and at 1 h after surgery, blood samples were obtained from the artery and jugular venous bulb for blood gas analysis and for calculation of jugular bulb blood oxygen content (CjvO 2), artery-jugular bulb blood oxygen content difference (Ca-jvO 2), cerebral oxygen uptake rate (CERO 2) and jugular-arterial blood lactate concentration difference (Djv-aLac). The emergence time, amounts of intraoperative anesthetics, use of noradrenaline, cumulative time of EEG burst inhibition and duration of AI<40 were recorded.The development of POD was assessed within 5 days after surgery by the confusion assessment method for the intensive care unit and the duration was recorded. Results:Compared with group C, recovery time, cumulative time of EEG burst inhibition and duration of AI<40 were significantly shortened, the intraoperative consumption of propofol and remifentanil was decreased, the requirement for intraoperative noradrenaline was increased, CjvO 2 was increased, Ca-jvO 2 and CERO 2 were decreased immediately at the end of surgery and at 1 h after surgery, the incidence of POD within 5 days after surgery was decreased, and POD duration was shortened in group E ( P<0.05). Conclusion:General anesthesia guided by EEG monitoring can reduce the development of POD in elderly patients with non-acute fragile brain function.

Chronic kidney disease (CKD) has become a worldwide health and social problem. Retarding its progression to end-stage renal disease is beneficial both to the patients and the healthcare system. Plenty of clinical trials have indicated that enema with Chinese medicine could effectively prevent chronic renal failure, and was widely used in the clinical practice. However, studies on mechanism were still nearly blank, which may prevent further improvement of therapeutic efficacy. Recent studies had discovered that colon was an important organ where uremic toxins were generated. The uremic toxins involved could not only promote CKD progression, but also was closely correlated with CKD mortality. Reducing production and promoting excretion of toxins were confirmed to reduce renal tubule interstitial fibrosis and delay renal progression. On the basis of the theory of gut-kidney axis above, we had conducted pilot clinical researches to evaluate the effect of enema with Chinese medicine on the intestinal flora, gut barrier, enterogenous uremic toxins and renal protection. The preliminary results revealed that rheum enema through colon could accelerate intestinal dynamics, improve intestinal barrier function, regulate intestinal flora and reduce production and absorption of intestine-derived uremic toxins such as indoxyl sulfate, which may reduce renal fibrosis and delay renal progression. Further studies could provide more evidence for colon as a new therapeutic target for the treatment of CKD with Chinese medicine.
Colon ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Enema ; Humans ; Kidney ; drug effects ; pathology ; Renal Insufficiency, Chronic ; drug therapy ; pathology ; Treatment Outcome

[Objective]To explore the effects of family cohesion and child-teacher dependency on children's attention-deficit/hy?peractivity behaviors at one year after their kindergarten entry.[Methods]Based on a cohort of 7465 first-year preschoolers in Long?hua district of Shenzhen in 2014,the information about social-demographic characteristics,family cohesion(assessed with Family Environment Scale-Chinese Version)were collected from their primary-caregivers at kindergarten entry. The teacher rating scores of child-teacher dependency(assessed with Student-Teacher Relationship Scale)and parental rating scores of attention-deficit/hyperac?tivity behaviors(assessed with Conners'Parent Symptom Questionnaire)were examined at one year after their kindergarten entry. Lo?gistic regression were performed to analyze the relationship and interaction between family cohesion and child-teacher dependency in attention-deficit/hyperactivity behaviors,after adjusting for potential confounders.[Results]Children's attention-deficit/hyperactivity behaviors at one year after kindergarten entry were negatively associated with family cohesion (OR=0.85,95% CI=0.75~0.97,P=0.01),but were not significantly associated with child-teacher dependency(OR=1.00,95%CI=0.93~1.08,P=0.98). Furthermore, family cohesion moderated the impact of child-teacher dependency on children ' s attention-deficit/hyperactivity behaviors at one year after kindergarten entry(OR=0.96,95% CI=0.92~0.99,P=0.03). In addition,neither family cohesion(OR=1.08,95%CI=0.88~1.34,P=0.47)nor child-teacher dependency(OR=1.03,95%CI=0.93~1.15,P=0.53)could protect preschoolers from persistence of attention-deficit/hyperactivity behaviors in the first year of kindergarten.[Conclusion]The association of child-teacher dependency and onset of attention-deficit/hyperactivity behaviors is modified by the level of family cohesion in preschool children.

AIMTo investigate the pharmacokinetics of hylotelephin in Beagle dogs and obtain the main pharmacokinetic parameters.

METHODSAn HPLC method with UV detection was developed to study the pharmacokinetics of hylotelephin in dogs by joining an internal standard (anthracene). Benzoyl chloride was used to the pre-column derivatization of hylotelephin and methanol-water (64:36) was used as the mobile phase. According to the 3P97 pharmacokinetic program, the main parameters were calculated.

RESULTSThe hylotelephin pharmacokinetics conforms to a two-compartment open model after a single iv dose of hylotelephin 10.6 or 21.3 mg x kg(-1) in Beagle dogs. The parameters of two groups were as follows: T(1/2) alpha were 2.3 and 2.1 min, T(1/2) beta were 1.9 and 2.0 h, K12 were 0. 12 and 0.11 min, K21 were 0.17 and 0.21 min, K10 were 0.011 and 0.0094 min, Vc were 0.54 and 0.54 L x kg(-1), AUC were 1.8 and 4.1 g x min x L(-1), CL were 0.0048 and 0.0056 L x kg(-1) x min(-1), MRT were 2.10 and 2.4 h, respectively.

CONCLUSIONThe pharmacokinetics of hylotelephin after iv administration showed a rapid distribution and elimination process in Beagle dogs and was of first order kinetics.

Animals ; Antiviral Agents ; chemistry ; isolation & purification ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; Crassulaceae ; chemistry ; Dogs ; Female ; Heterocyclic Compounds, 3-Ring ; chemistry ; isolation & purification ; pharmacokinetics ; Male ; Molecular Structure ; Plants, Medicinal ; chemistry ; Spectrophotometry, Ultraviolet

OBJECTIVETo detail our early experience and technique of a modified two-stage reimplantation protocol using antibiotic-loaded articulating cement spacers (ALACSs) for treatment of late periprosthetic infection after total knee arthroplasty (TKA).

METHODSFrom January 2006 to February 2009, a series of 21 patients (21 knees) with late infected TKAs were treated by radical debridement and removal of all components and cement, and then articulating spacers were implanted using antibiotic-impregnated bone cement. For this purpose, 4 g vancomycin powder was mixed with per 40 g cement. Graduated knee motion and partial weight bearing activity were encouraged in the interval period. Each patient received an individual systemic organism-sensitive antimicrobial therapy for 4.9 (range, 2-8) weeks followed by a second-stage TKA revision. All the patients were regularly followed up using the American Knee Society Scoring System.

RESULTSEach case underwent a successful two-stage exchange and had infection eradicated, none had recurrent infection after an average of 32.2 (range, 17-54) months of follow-up. Preoperatively, the mean knee score was 53.5 points, function score was 27.3 points, pain score was 25.7 points, range of motion (ROM) was 82.0 degree extensor lag was 2 degree Between stages, the mean knee score was increased to 61.3 points, function score to 45 points, pain score to 35 points, ROM to 88.2 degree and extensor lag to 3.4 degree At final follow-up, the mean knee score was further increased to 82.1 points, function score to 74.5 points, pain score to 42.1 points, ROM to 94.3 degree and knee extension lag to 1.9 degree The interval period was 11.5 (range, 6-32) weeks. The amount of bone loss was unchanged between stages. No patient developed noticeable dysfunction of the liver or kidney or other complications such as impaired wound healing, deep venous thrombosis, pulmonary embolism, cerebrovascular accidents, etc.

CONCLUSIONSTreating infected TKA with ALACS avoids spacer-related bone loss, preserves knee function between stages, and eradicates infection effectively without significant complications. The early clinical results are inspiring. The authors believe that radical and repeated (if needed) debridement, individual application of systemic antibiotics, and reasonable timing judgement upon the secondary revision are all key factors related to a successful outcome with two-stage reimplantation procedure for infected TKA.

Anti-Bacterial Agents ; Arthroplasty, Replacement, Knee ; Humans ; Knee Joint ; Knee Prosthesis ; Prosthesis-Related Infections