OBJECTIVETo investigate the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid (HA) on the proliferation of rabbit chondrocytes in vitro.

METHODSChondrocytes from the knee joints of New Zealand white rabbits were cultured. bFGF or HA or both were added into the culture medium respectively, and the proliferation of the chondrocytes was measured with MTT 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyl-tetra-zolium bromide. (MTT, Sigma, M2128).

RESULTSBasic fibroblast growth factor (10 ng/ml) with low concentration of fetal bovine serum in the culture medium promoted the proliferation of chondrocytes significantly, and this effect reached its maximum when concentration of bFGF reached 50 ng/ml. HA itself had no effect on the proliferation of chondrocytes. However, when bFGF was used in combination with HA, especially when the concentration of bFGF was 50-500 ng/ml and that of HA was 10-50 ng/ml, the effect on the proliferation of chondrocytes was much more than when bFGF or HA was used alone.

CONCLUSIONSbFGF can promote the proliferation of chondrocytes. HA, which has no effect on the proliferation of the cells, can maintain a normal growth of chondrocytes. When bFGF is used in combination with HA, more proliferation is obtained.

Analysis of Variance ; Animals ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Chondrocytes ; drug effects ; physiology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Hyaluronic Acid ; pharmacology ; In Vitro Techniques ; Knee Joint ; cytology ; Male ; Probability ; Rabbits ; Sensitivity and Specificity

OBJECTIVETo analyze the clinical features of mid- and long-term acute cardiac allograft rejection to improve the long-term clinical outcomes of the patients.

METHODSFourteen recipients (11 males and 3 females) underwent orthotopic heart transplantation with standard immunosuppressive therapy protocols (3 cases) or induction therapy protocols (11 cases). Cyclosporine, azathioprine or mycophenolate mofetil, and prednisolone were applied as the maintenance immunosuppressive regimen. Acute graft rejection episodes occurred within 3 to 6 months in 1 case, within 6 months to 1 year in 3 cases, within 1 to 2 years in 3 cases, within 2 to 5 years in 6 cases, and above 5 years in 1 case.

RESULTSNo significant difference was found in the incidence of late heart rejection between the patients receiving the two immunosuppressive therapy protocols. Immunosuppressants were withdrawn or spared in 8 recipients due to different causes. Nine recipients with steroid-sensitive acute cardiac allograft rejection were treated with steroid-pulse therapy, while the other 5 were treated with a short course of polyclonal antithymocyte antibodies because of steroid-resistant acute rejection; in 11 cases, azathioprine was converted to mycophenolate mofetil. Four of the 5 late deaths occurred in the recipients with steroid-resistant rejection. The surviving recipients had a good quality of life, and no recurrent episodes of rejection or infection were observed in the follow-up period.

CONCLUSIONSLate acute cardiac allograft rejection is associated mainly with patient compliance but not with early immunosuppressive therapy protocols. The episodes are rather severe and should be timely treated with steroid pulses or polyclonal antithymocyte antibodies.

Adolescent ; Adult ; Cyclosporine ; Female ; Graft Rejection ; etiology ; prevention & control ; Heart Transplantation ; adverse effects ; Humans ; Immunosuppression ; methods ; Male ; Middle Aged ; Mycophenolic Acid ; analogs & derivatives ; Young Adult

The purpose of this investigation is to improve ethanol production and decrease acetate formation in Saccharomyces cerevisiae strain YS2-?adh2.The strain YS2-?adh2 with deleted alcohol dehydrogenase Ⅱ(adh2) gene was isolated in our lab with higher ethanol production than that of the strain YS2.The ace-taldehyde dehydrogenase Ⅵ(ald6) gene encoded a cytosolic acetaldehyde dehydrogenase,a key enzyme of the pyruvate dehydrogenase(PDH) bypass,transfers acetaldehyde to acetate.To disrupt ald6 gene of the strain YS2-?adh2,ald6 gene targeting cassettes were synthesized by long flanking homology PCR(LFH-PCR) and then were transformed into YS2-?adh2 mutants by LiAc/SS Carrier DNA/PEG method.Positive transformants were selected with G418 and further confirmed by PCR.Once correctly integrated into the genome,the selective marker was rescued by transforming the plasmid pSH65 into the positive transformants and inducing the Cre expression with a Cre/loxP-mediated marker removal procedure.We named the ald6 gene knocked-out strain as YS2-?adh2-?ald6 which has a 12.5% higher ethanol production and a 18% lower acetate formation compared to the strain YS2.

Objective To study the accuracy of real-time continuous monitoring system (RT-CGMS) at different stages and its association with glucose excursion. Methods Totally 33 patients with type 1 diabetes or type 2diabetes were under surveillance of RT-CGMS for 5 d. Capillary glucose values were measured 7 times daily.Correlation coefficient, error grid analysis (EGA), and Bland-Altman analysis methods were used to assess the correlation, accuracy and agreement of RT-CGMS at different stages and in general level; The mean amplitude of glucose excursion (MAGE) and the frequency of glucose excursion ( FGE ) were also calculated. Results ( 1 ) The correlation coefficient of RT-CGMS with capillary glucose values at fasting, postprandial stages, and in general level were 0.94,0.92, and 0.93 respectively( P＜0.01 ). (2) EGA showed that 98.82%, 98.39%, and 98.64% of the results fell in the A and B zones and 1. 18%, 1.61%, and 1.36% fell in the D zone respectively at fasting,postprandial stages, and in general level. There is no result fell in C and E zones. ( 3 ) The agreement analysis showed that RT-CGMS readings were in close agreement with capillary glucose values at fasting, postprandial periods, and in general level. (4)The MAGE at fasting, postprandial periods, and in general level were (3.57±2.66), (4.07±3.09), and (4. 02 ±3.04) mmol/L (P＞0. 05), (0±0. 5), (3± 1), and( 1 ±3) d for FGE (P＜0. 01 ).Conclusion RT-CGMS at fasting stage has higher accuracy than postprandial stage and general level, FGE at fasting stage is higher than postprandial stage and general level.

Objective To analyze the safety of indwelling neonatal umbilical vein catheter (UVC) at high or low tip positions.Methods We retrospectively analyzed 155 neonates with indwelling UVC in Guangdong Women and Children Hospital from March 2015 to May 2016.According to the position of the catheter tip,these infants were divided into high position group (the tip was at the same level as the central vein) or low position (the tip was below the portal vein) group.Several parameters including indwelling time,liver function,liver B-ultrasound and catheter-related bloodstream infections in the two groups were analyzed.Data were statistically analyzed using Mann-Whitney U test,Wilcoxon signed rank sum test or Chi-square test.Results (1) A total of 155 infants were enrolled in this study.Their gestational age ranged from 25 to 41 weeks and their birth weight were 700 g to 4 690 g.UCV was inserted at the age of 0.5-8.0 d for 0-12 d.In the low position group,seven cases ended in early extubation due to fluid extravasation caused by shallow insertion.The indwelling time of the low position group was shorter than that of the high position group [M (P25-P75),6 (4-7) d and 7 (5-7) d,Z=-2.580,P=0.010].There were no significant differences in gender,gestational age,birth weight,and age at catheterization between the two groups.(2) No abnormality in the two groups was revealed by liver ultrasound.Complications such as neonatal necrotizing enterocolitis,thrombosis and embolism,air embolism and liver abscess were not reported.The proportion of liver function abnormalities in the high group was 6.9％ (6/87),which was not statistically significant as compared with 4.4％ (3/68) in the low position group (3x2=0.431,P=0.512).(3) Blood culture was performed for all cases,of which 116 (74.8％) were catheter culture (including 71 in the high position group and 45 in the low position group).The incidence of catheter-related bloodstream infection in the high position group was 5.6％ (4/71) and the infection rate was 7.4 per thousand catheter days,while in the low position group these figures were 6.7％ (3/45) and 8.0 per thousand catheter days,respectively.There was no significant difference between the two groups (x2=0.052,P=0.820).(4) Results of catheter culture showed that seven cases were positive,including three for Staphylococcus haemolyticus (two in low position group and one in high position group),one for yeast-like fungus (high position group),one for Enterococcus faecium (high position group),one for Staphylococcus aureus (high position group) and one for Candida albicans (low position group).Conclusions The indwelling time of UVC at high tip position is longer than that at low position,and although there are no differences between the complications and the incidence of catheter-related bloodstream infection in the two groups,but its safety needs further study.

Cell Line, Tumor

OBJECTIVETo investigate the outcome of orthotopic heart transplantation for patient with end-stage hypertrophic cardiomyopathy.

METHODSThis retrospective review analyzed the clinical data of nine patients (7 males) undergoing orthotopic heart transplantation for end-stage hypertrophic cardiomyopathy in our center. All patients received induced therapy protocols peri-operative and standard triple maintenance immunosuppressive therapy postoperative.

RESULTSOne recipients developed acute renal failure due to renal artery embolism and allograft rejection in the early posttransplantive course, symptoms and signs were improved under continuous renal replacement therapy and steroid-pulse therapy, this patient died of sudden cardiac arrest at 32 months post transplantation. Another recipient developed demyelinating disease in frontal and parietal lobe and finally recovered with medical therapy. Eight patients survived the operation with good quality of life and there was no episode of rejection or infection or chronic graft arteriosclerosis during follow-up time. Three recipients developed left ventricular hypertrophy and there were no signs of grapg-vessel diseases in the survivals.

CONCLUSIONHeart transplantation is the best therapeutic option for selected patients with end-stage hypertrophic cardiomyopathy.

Adolescent ; Adult ; Cardiomyopathy, Hypertrophic ; surgery ; Female ; Heart Transplantation ; Humans ; Male ; Middle Aged ; Postoperative Period ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo detect HPV 58, a common type of human papillomavirus (HPV), clone and express its E7 gene from biopsy specimens of cervical cancer.

METHODSHPV 58 from 58 biopsy tissues of cervical cancer was detected by GP5+/GP6+ PCR followed by template-directed dye-terminator incorporation assay with fluorescence polarization detection (TDI-FP). HPV 58 E7 gene was amplified from one HPV 58-positive sample, and then cloned into pGEM-T Easy vector. The recombinant plasmid, HPV58E7-pGEM-T was confirmed by sequencing. Subsequently, E7 gene was cloned into prokaryotic expression vector pRSET-A. The constructed pRSET-58E7 plasmids were transfected into BL21(DE3) cells, and induced to express 58 E7 protein by IPTG.

RESULTSAmong the 58 biopsy tissues of cervical cancer, 10 were HPV 58-positive, accounting for 19.2% of 52 HPV-positive cases. HPV 58 E7 gene was amplified from one HPV 58-positive sample. The constructed plasmids were identified containing HPV58 E7 gene by restriction enzyme analysis and sequencing. SDS-PAGE analysis showed that HPV58 E7 His6 fusion protein of M(r) 16 x 10(3) was expressed by pRSET-58E7 after induction by IPTG. The fusion protein accounted for 30% of total bacterial proteins.

CONCLUSIONHPV 58 is not uncommon in Chinese women with cervical cancer in Shaanxi province. Constructed HPV58 E7 recombinant plasmids can be effectively expressed in E.coli, which may provide a tool in diagnosis and vaccine design for HPV of HPV58-associated tumors.

Adult ; Cloning, Molecular ; Escherichia coli ; metabolism ; Female ; Genes, Viral ; Humans ; Middle Aged ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus E7 Proteins ; biosynthesis ; genetics ; Papillomavirus Infections ; complications ; genetics ; Plasmids ; Recombinant Proteins ; biosynthesis ; genetics ; Transformation, Genetic ; Uterine Cervical Neoplasms ; genetics ; virology

OBJECTIVETo evaluate the clinical outcomes between extreme lateral interbody fusion and conventional posterior operation in the treatment of upper lumbar disc herniation.

METHODSAmong 60 patients with upper lumbar disc herniation were treated with extreme lateral interbody fusion(XLIF) or conventional posterior operation from June 2010 to December 2014, 30 patients(19 males and 11 females) were treated with XLIF (XLIF group); and the other 30 patients(17 males and 13 females) were treated with conventional posterior operation (conventional group). In XLIF group, the lesions occurred at T₁₂L₁ segments in 2 patients, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 10 patients, and at L₃,₄ segment in 12 patients. In conventional group, the lesions occurred at T₁₂L₁ segments in 1 patient, at L₁,₂ segments in 6 patients, at L₂,₃ segments in 8 patients, and at L₃,₄ segment in 15 patients. Operative incision lengths, time, blood loss, postoperative draining volume, hospital stays were recorded. Pre-and post-operative visual analogue score(VAS) and Japanese Orthopedic Association(JOA) were compared between two groups. According to the image data, the intervertebral fusion device was observed to be displaced and the rate of interbody fusion was analyzed.

RESULTSAll the patients were followed up, and the duration ranged from 12 to 48 months, with an average of 29 months. The complications included 2 femoral nerve damage in XLIF group (postoperative recovery within 3 months) and superficial incision infection in conventional group(cured by anti-infection). There were no patients with cerebrospinal fluid leakage(CSFL), cauda equina injuries or functional deterioration in the nerve root of lower limbs. In the XLIF group: the operative time was (65.6±20.5) minutes, blood loss was (48.8±15.3) ml, postoperative draining volume was 0 ml. In the conventional group: the operative time was (135.2±33.9) minutes, blood loss was (260.3±125.7) ml, postoperative draining volume was (207.1±50.2) ml. The operative time, blood loss, postoperative draining volume in XLIF group were less than those in the conventional group(<0.05). The JOA and VAS score were significantly improved in both groups during the follow-up period compared with those before operation(<0.05). But the difference of the JOA and VAS score between the two groups 1, 6, and 24 months after surgery had not significant differences(>0.05). There were no significant differences in the fusion rate between the two groups 6 and 12 months after operation(>0.05).

CONCLUSIONSThe XLIF fusion for the treatment of upper lumbar disc herniation has several advantages such as minimal invasive, stable vertebral plate, less complications and postoperative fusion rate, which has a better clinical effect.

To study the program of evaluating mothers and infants after 6 months of cord blood donation, from June 1998 to February 2004, all mothers after 6 months of cord blood donation were followed-up by phone calls or letters to report on the health condition. The results showed that when 3 195 mothers were visited by phone calls, 18 mothers declined to answer. 392 letter were send to those who could not be found by phone, 15 of whom wrote back. The average time to talk with each mother was approximately 12 minutes. Follow-up on the baby donors showed two cases with chromosome abnormality, one with hypothyroidism, one with neutropenia, one with albinism and 5 dead with unclear reasons. The cord blood components from all these abnormal donors found were discarded. In conclusion, the programs to evaluate mother and baby after 6 months of cord blood donation seems important in quality control of the components stored in cord blood bank.
Adult ; Blood Banks ; Blood Donors ; China ; Female ; Fetal Blood ; Follow-Up Studies ; Humans ; Infant ; Infant, Newborn ; Quality Assurance, Health Care ; methods ; statistics & numerical data ; Surveys and Questionnaires ; Time Factors