0.05).The DNA content in samples of ITS group was significantly lower than that of FBS group on day 6(P

BACKGROUND:Self-assembling short-peptide has been shown to be a kind of hovel nanometer biomaterial.Following culture combined with chondrocytes,self-assembling short-peptide can promote eell marex secretion,cell division and proliferation.and eell phenotype maintenance.For this reason,it has become a candidate for all ideal cell scaffold used for cartilage tissue engineering.OBJECTIVE:To observe the biological characteristics of rabbit articular chondrocytes cultured in three-dimensional nanoscale self-assembling short.peptide KLD-12 hydrogel seaffold.DESIGN.TIME AND SETTING:An observational experiment was performed at the laboratory of Institute for Nanobiomedical Technology and Membrane Biology,Sichuan University between November 2007 and April 2008.MATERIALS:Nanoscale short-peptide KLD-12 with a sequence of Ac-KLDLKLDLKLDL-CONH2 was synthesized by Shanghai Biotech Company,China.METHODS:Chondrocytes were harvested from New Zealand rabbit articular cartilage by enzyme digestion.Following in vitro culture.chondrocytes of passage 3 were inoculated into 4 g/L self-assembling short-peptide KLD-12 solution at a density of 5x 109/L.Peptide gel formation was induced by phosphate buffered saline for three-dimensional culture.MAIN OUTCOME MEASURES:①Observation of chondrocyte morphology in the short-peptide hydrogel with inverted phase contrast microscope.②Detection of intracellular matrix secretion by toluidine blue staining and immunohistochemistry.③Measurement of mRNA expression levels of aggrccan,type Ⅱ collagen and type Ⅰ collagen in chondrocyte-seeded hydrogels by reverse transcription-polymerase chain reaction.RESULTS:①The viability of chondrocytes digested and isolated from rabbit articular cartilage was over 95%.④Chondrocytes cultured in three-dimensional short-peptide hydrogel maintained their round shape,proliferated actively,and aggregated in clusters.In addition.cartilage lacuna could be seen surrounding the clusters.③Both toluidine blue staining and type Ⅱ collagen immunohistochemistry results were positive.The mRNA expression levels of aggrecan and type Ⅱ collagen were increased with time.④Reverse transcription-polymerase chain reaction results revealed that after 3 weeks of in vitro culture in the short-peptide hydrogel,chondrocytes maintained the capability to secrete aggrecan and to express type Ⅱ collagen.CONCLUSION:Self-assembling short-peptide KLD-12 three-dimensional hydrogel Call well inainrain the normal cellular morphology,function and proliferative capability of chondrocytes

BACKGROUND: In the tissue engineering studies, the common method of cell detachment is trypsinization. However, this method may decrease cell activity by breaking the extracellular matrix proteins. Therefore, it is important to develop a new cell detachment method which can avoid the damage from trypsinization.OBJECTIVE: Based on that poly N-isopropylacrylamide (PNIPAAm) could alter its hydrophobic property with the change of temperature, this study aimed to develop a novel method which can control cell detachment by simply changing the temperature. METHODS: IPAAm was dissolved in isopropanol with final concentration of 55% (w/v). IPAAm solution was evenly laid on the surface of TCPS dish, and PNiPAAm was then covalently grafted on the TCPS dish surface by electron beam irradiation. The micro.structure of the surface of TCPS dish was observed by atomic force microscopy (AFM) before and after PNIPAAm grafted. Water contact angle was determined by sessile drop method. And an inverted phase contrast microscope was used to observe and evaluate the cell detachment ability of PNIPAAm grafted TCPS dish by subjecting the same dish to different temperatures. RESULTS AND CONCLUSION: AFM images showed that a thin and compact PNIPAAm layer formed on the TCPS dish surface. In the water contact angle test, there was no statistic difference between PNIPAAm grafted and ungrafted surfaces at 37 ℃, and the water contact angle of PNIPAAm grafted surfaces was lower than that of ungrafted surface at 20 ℃. The cells adhered to the surface of the PNIPAArn grafted TCPS dish at 37 ℃, which would be detached at 20 ℃.It was successful to control cell detachment by simply changing the temperature with PNIPAAm grafted TCPS dish, and it would be promising to be applied in tissue engineering.

Objective To study the clinical features of congenital glucose-galactose malabsorption (CGGM),and to improve the understanding of CGGM.Method Clinical manifestations and treatment process of one patient with CGGM in our hospital were retrospectively analyzed.From 1966 to 2016 May,Chinese medical database and PUBMED were searched using Malabsorption syndrome,dehydration,hypernatremia , diarrhea , newborn , carbohydrate metabolism ,andglucose/galactose malabsorption as key words.The clinical features of CGGM reported in literatures were summarized.Result The patient in our hospital was a full-term female infant naturally delivered.The onset of the disease was on the 9th day after birth,and the clinical manifestations included severe diarrhea,severe dehydration,hypernatremia,metabolic acidosis and malnutrition.After intravenous infusion and symptomatic treatment,dehydration,hypernatremia and metabolic acidosis were corrected.However,there was no improvement of diarrhea characterized with watery and acidic stools,and neither was weight gain.Glucose loading test was negative,and fructose loading test was positive.Diarrhea was improved markedly using diagnostic carbohydrate-free formula,so CGGM was diagnosed clinically.SLC5A1 homozygous IVS7-2 A ＞ G mutation was detected which confirmed the diagnosis of CGGM.With carbohydrate-free formula feeding,the body weight of the infant was increased.Followed up for 2 months now,her body length and body weight were at P25 and P22 on growth curve respectively,and no obvious neurological sequela was observed.Our literature review revealed 7 reports including 48 cases of CGGM children.Literature review showed that:most children with CGGM (79.2％) had the onset within 7 days of life;main clinical features included diarrhea (100％),dehydration (100％),and malnutrition (54.2％);22.9％ of patients with carbohydrate-free formula and 27.1％ with fructose matrix formula were fed well;no death was detected,77.1％ had normal weight gain,and 91.7％ had normal development of the nervous system.Conclusion CGGM is rare.The symptoms include severe watery and acidic stools with onset during neonatal period.CGGM is associated with severe complications such as hypertonic dehydration and hypernatremia.The diagnosis is established based upon typical clinical manifestations,sugar loading test and SLC5A1 gene detection.Carbohydrate-free formula feeding is effective.

Objective:To develop the Counseling Skills Rating Scale for Psychological Aids Hotline (CSRSPAH),an instrument to objectively assess the quality of process of counseling or intervention provided by hotline operators,and to test its validities and reliabilities.Methods:A draft of the scale,which consisted of 50 items under 5 dimensions,was designed in line with previous researches and the practice.In pilot study,supervisors at Beijing psychological aids hotline,assessed tape recorded sessions and gave feedbacks on the draft of the scale.The scale had been revised based on the pilot study.Twenty-eight items were deleted,and several items were rephrased.Finally,a scale which consisted of 22 items within 3 dimensions,i.e.counseling process,attitude,and communication skill,was developed.The scoring standards of the scale were also developed.To evaluate the IntraClass Coefficients (ICC) of the CSRSPAH,each of the tape records of 37 callings from 2005 to 2007 were assessed by 7 supervisors independently,using the scale.And the tape records of other 318 callings from 2013 to 2014 were also assessed by supervisors,using the CSRSPAH.The results of the 318 assessed callings were used to test the construct validities with the Confirmative Factor Analysis.The Cronbach a coefficients of the total score and three dimensions,the discriminant indices of every items,and correlations of each items and each dimensions were calculated,based on the 318 assessed callings.Results:One of the items (referral) was deleted due to excessive amount of missing data.Results of cortfirmative factor analysis of the remained 21-item scale revealed that the 3-factor construct structure of the scale was robust.The fitting indices of the confirm factor analysis were,x2/df=675.21/186,CFI =0.92,NNFI =0.91,RMSEA =0.10,SRMR =0.08.The Cronbach α coefficients of the total score,scores of counseling process,attitude,and communication skill,were 0.89,0.68,0.81 and 0.77,respectively.The ICCs of the inter-rater reliabilities of the total score and 3 dimensions of the scale were 0.67,0.59,0.59,and 0.67,respectively.The discriminant indices of all the 21 items ranged from 0.09 to 0.32.The correlation coefficients of scores of each items and scores of 3 dimensions and total scores were greater than 0.30,and reached statistical significance.Conclusion:The validities and reliabilities of the Counseling Skills Rating Scale for Psychological Aids Hotline are acceptable.The scale could be used in assessing the quality of hotline counseling or intervention,and related studies in the future.

Objective To evaluate the lung protection of remote limb ischemic preconditioning during one-lung ventilation (OLV) in the patients undergoing esophageal cancer resection.Methods Seventyone patients of both sexes,aged 30-64 yr,with body mass index of 15-28 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective esophageal cancer resection,were randomly divided into control group (group C,n =34) and remote limb ischemic preconditioning group (group RLIP,n =37) using a random number table.Patients in group RLIP received three cycles of 5-min ischemia/5-min reperfusion induced by a blood pressure cuff placed on one upper arm before OLV.Before OLV (T0),at 1 and 2 h of OLV (T1,2),at 20 min after re-expansion of the collapsed lung (T3),and at 2 h after operation (T4),blood samples were drawn from the radial artery for blood gas analysis,oxygenation index (PaO2/FiO2) and alveolar-arterial oxygen gradient (A-aDO2) were calculated.At T0,T2,T3 and T4,blood samples were collected from the radial artery for determination of plasma tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6),and IL-10 concentrations.Results Compared with group C,PaO2/FiO2 was significantly increased,and A-aDO2was decreased at T1,2,the plasma TNF-α concentrations were decreased at T2-4 (P＜0.05),and no significant change was found in the plasma IL-6 and IL-10 concentrations and rate of abnormal pulmonary function at T1-4 in group RLIP (P＞O.05).Conclusion Although remote limb ischemic preconditioning can produce lung protection during OLV in the patients undergoing esophageal cancer resection,it provides no clinical significance.

Organic chemistry experiment is early and important practical course for pharmacy speciality after entering college,which is very important for training their practical abilities and spirit of innovation.The method of integrating four autonomies (self-designing,teaching,operating and summarizing) has been tried in organic chemistry experiment teaching,which completely takes the student as the center and puts emphasis on inquiry learning and ability training.The preliminary practice shows that it is of benefit to arousing students' enthusiasm for study,training their ability and developing their innovative consciousness and spirit.

Objective To evaluate a new gene therapy for the treatment of experimental occlusive arterial disease Methods Magnetic nanospheres were produced, VEGF gene was cloned for subsequent construction of eukaryotic expression plasmid The magnetic gelatin microspheres used in targeted gene therapy were prepared by emulsion crosslinking method The microspheres were injected intrafemorally in rabbits through contralateral femoral artery, and the ischemic limb was placed in a magnetic field Angiography was performed on day 10 and day 30 respectively The capillary density and the capillary to muscle fiber ratio were determined histochemically Results Compared to the controls there was significant collateral artery development in VEGF transfected group The capillary density and the capillary to muscle fiber ratio were significantly higher for the VEGF transfected group than for the control group. The capillary density of control was (125?23)/mm 2, and in VEGF group was (298?27)/mm 2, P

ABSTRACT:In the present study ,we aimed to identify differentially expressed proteins between induced worms (the infec‐ted mice were treated intragastrically with ED50 PZQ) and uninduced worms (control group) for clarifying the mechanism of PZQ .ED50 PZQ was used to administrate mice that were infected with S .japonicum via intragastric incubation for consecutive‐ly 30 days .Twenty‐one days later ,mice were sacrificed after treatment with 200 mg/kg PZQ for continuously five days ,and the male worms were obtained and some of them were subjected in DMEM medium with different concentrations of PZQ in vitro for 16 hours .Then the worms were washed twice and incubated in PZQ‐free medium for 72 hours .Compared with control group ,the induced worms had lesser sensitivity to PZQ .The survival rate of induced worms was 75 .6% in vitro when the con‐centration of PZQ was 112 mol/L (the concentration was 8 times of uninduced worms Lethal Concentration ) ,significantly higher than that in the uninduced worms (11 .1% ,P<0 .05) ,showing obviously tolerance .The other induced and uninduced worms were acquired and collected for 2D‐DIGE and MALDI‐TOF‐MS ,and combined with bioinformatics to analyse the func‐tion of the identified protein .Thirty differential expression proteins were confirmed between induced and uninduced worms ,in‐cluding 12 proteins up‐regulated and 18 proteins down‐regulated .These proteins respectively ascribed to cytoskeleton‐associat‐ed protein ,glucose and energy metabolism enzymes ,stress proteins ,thioredoxin peroxidase enzymes ,and other protease .Up‐or down‐regulation of these differential proteins indicated that PZQ promote or inhibit the expression of some specific genes . These findings may help to clarify the mechanism of PZQ ,simultaneously ,providing a scientific basis for exploring new vaccine candidate antigens and targets for drug therapy .

Objective To investigate the relationship between adiponectin combined with the ultrasound blood flow index of the umbilical artery and perinatal outcome in women with severe preeclampsia. Methods Placental tissues were obtained from normal term pregnancies (control group, n=50) and severe preeclampsia patients (PE group, n=50) in Third Affiliated Hospital of Zhengzhou University from February 2014 to October 2014. The expression of adiponectin was examined using immunohistochemical methods and real-time polymerase chain reaction. The umbilical artery was measured by color Doppler, and the umbilical artery systolic/diastolic ratio (UA-S/D), umbilical artery resistance index (UA-RI) and umbilical artery pulsatility index (UA-PI) were determined. The relationship between the expression of adiponectin in placental tissues, UA-S/D and perinatal outcome were analyzed. The data were analyzed using two dependent-sample t test, the log-rank test and Spearman correlation analysis. Results Compared with the control group, infants in the PE group had lower birth weight and placental weight, shorter height, and greater umbilical artery indices including UA-S/D, UA-RI and UA-PI (all P<0.05). The expression of adiponectin and its mRNA in placentae of the PE group was significantly higher than that of the control group (adiponectin: 0.326±0.011 vs. 0.116±0.011, t=99.144, P=0.000;mRNA:4.18±1.80 vs. 1.00±0.51, t=11.985, P=0.000). UA-S/D had a negative correlation with birth weight, onset gestational age and gestational age at birth (r= - 0.897, - 0.469 and - 0.524, all P<0.01). The expression of adiponectin mRNA had a negative correlation with birth weight, onset gestational age, and gestational age at birth (r=-0.580,-0.407 and-0.449, all P<0.01). The expression level of adiponectin had positive correlations with body mass index of the mothers and the UA-S/D (r=0.261 and 0.788, both P<0.01). Conclusions The expression of adiponectin in placental tissues and blood flow index of the umbilical artery both increase in severe preeclampsia, and are associated with poor perinatal outcome.