OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.

METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.

RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).

CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.

Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

BACKGROUND: The size of artificial disc is not exactly identical to cervical vertebral endplate. However, the effect of implant size on cervical disc replacement has not yet been clearly identified. OBJECTIVE: To evaluate the effect of Bryan disc size on cervical artificial disc replacement. METHODS: Totally 71 patients with cervical degenerative disease underwent single-level Bryan disc replacement from December 2003 to December 2007 were enrolled. The average age was (45.90±8.12) years old, and all patients were followed up for more than 10 years. The artificial disc size was measured using lateral X-ray films, and expressed as (Bryan artificial disc's footprint sagittal diameter×2) / (upper endplate sagittal diameter + lower endplate's sagittal diameter)×100%. X-ray films were photographed to measure motion range of the treated segment before surgery, 3 months after surgery, and during final follow-up. Clinical outcomes, including Japanese Orthopaedic Association score, Neck Disability Index and Odom's scores, were evaluated before and after operation. Observational indexes of different disc sizes (≥95% and <95% groups) were compared and analyzed. RESULTS AND CONCLUSION: (1) There were 49 patients in the ≥95% group and 22 patients in the <95% group. (2) At 3-month follow-up, range of motion was significantly smaller in the <95% group than in the ≥95% group (P < 0.01). At the final follow-up, range of motion was significantly smaller in the <95% group than in the ≥95% group (P < 0.05). (3) At final follow-up, improvement rate of Japanese Orthopaedic Association score was (70.65±32.58)% in the <95% group and (68.83±38.85)% in the ≥95% group, and no significant difference was detected between the two groups (P > 0.05). (4) At final follow-up, Neck Disability Index decreased by (10.82±7.50)% in the <95% group, and (12.61±8.51)% in the ≥95% group, and no significant difference was detected between the two groups (P > 0.05). (5) At final follow-up, Odem's score results showed excellent in 10 cases (45%), good in 9 cases (41%) and average in 3 cases (14%) in the <95% group; and excellent in 26 cases (53%), good in 20 cases (41%) and average in 3 cases (6%) in the ≥95% group, and no significant difference was detected between the two groups (P > 0.05). (6) Results suggest that different sizes of artificial disc replacement have obtained good functional recovery, but small artificial disc may impact the range of motion. We should try to avoid placing the artificial disc smaller than 95%.

OBJECTIVETo identify hereditary nonpolyposis colorectal cancer (HNPCC) families based on the germline mutations of MLH1 and MSH2 mRNA.

METHODSRNA was extracted from the peripheral blood of the 14 members from 12 different families fulfilling Amsterdam Criteria II. The germline mutations of MLH1 and MSH2 mRNA were detected by cDNA sequencing analysis following reverse transcription-PCR(RT-PCR) with special primers, heat-resistance reverse transcriptase, and expand long template PCR. DNA was extracted from the peripheral blood of the 14 members, the corresponding exons, in which mutations were found using the above method, were amplified with Taq enzyme, sequencing analysis was followed.

RESULTSSix germline mutations were detected and identified from the 6 different families based on mRNA, 4 of them to be in MLH1, the other 2 in MSH2. The MLH1 mutations distribute in the exon 8, 12, 16, and 19. The MSH2 mutations distribute in exons 1 and 2. The 6 mutations were identified from the corresponding exons respectively in genomic DNA sequencing analysis. The mutation types involve in 4 missense, 1 silent, and 1 non-coding area mutations. Five out of the 6 mutations have not been reported previously. Five out of the 6 mutations were pathological, involving in 5 different families. The five families were identified to HNPCC families.

CONCLUSIONHNPCC family can be identified with RNA-based sequencing of MLH1 and MSH2 from peripheral blood, which has the advantages of both cost, time saving and high sensitivity.

Adaptor Proteins, Signal Transducing ; Biomarkers, Tumor ; genetics ; Carrier Proteins ; genetics ; Colorectal Neoplasms, Hereditary Nonpolyposis ; diagnosis ; genetics ; Female ; Germ-Line Mutation ; Humans ; Male ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; genetics ; Mutation ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; genetics ; RNA, Messenger ; analysis

OBJECTIVETo introduce a new sequential chlorination disinfection process in which short-term free chlorine and chloramine are sequentially added.

METHODSPilot tests of this sequential chlorination were carried out in a drinking water plant.

RESULTSThe sequential chlorination disinfection process had the same or better efficiency on microbe (including virus) inactivation compared with the free chlorine disinfection process. There seemed to be some synergetic disinfection effect between free chlorine and monochloramine because they attacked different targets. The sequential chlorination disinfection process resulted in 35.7%-77.0% TTHM formation and 36.6%-54.8% THAA5 formation less than the free chlorination process. The poorer the water quality was, the more advantage the sequential chlorination disinfection had over the free chlorination.

CONCLUSIONThis process takes advantages of free chlorine's quick inactivation of microorganisms and chloramine's low disinfection by-product (DBP) yield and long-term residual effect, allowing simultaneous control of microbes and DBPs in an effective and economic way.

Bacteria ; drug effects ; Chloramines ; chemistry ; Chlorine ; chemistry ; Disinfection ; methods ; Viruses ; drug effects ; Water ; chemistry ; Water Microbiology ; Water Purification ; methods

OBJECTIVETo carry out a genetic detection and analysis of Von Hippel-Lindau (VHL) gene in Chinese patients with sporadic pheochromocytoma.

METHODSDNA samples were extracted from peripheral blood cells and fresh pheochromocytoma specimens from 41 patients with sporadic pheochromocytoma were assayed by polymerase chain reaction and direct sequencing. The DNA samples of 50 healthy volunteers were extracted from peripheral blood as a control. The PCR products of exon 1, exon 2 and exon 3 were used for molecular analysis of the VHL gene. The genetic detection of family members of VHL gene mutations was also performed.

RESULTSOne of mutations was located at nucleotide 572 (G-->C) in exon 2, presenting a codon 120 from arginine (R) to threonine (T). Tow small insertions were locatated at nucleotide 623T (TTTGTtG) in exon 2, leading to a frameshift mutation. There were also three carriers of G572C and three carriers of 623T (TTTGTtG) in family members of the three cases.

CONCLUSIONThere are some Chinese patients with sporadic pheochromocytoma with tumorigenic VHL gene mutations. It is recommended to use the genetic detection and analysis of VHL gene as a routine examination for patients with sporadic pheoehromoeytoma under the age of 50 years with questionable family history. The genetic detection and analysis of VHL gene may be useful as a marker for the diagnosis of hereditary pheochromocytoma.

Adrenal Gland Neoplasms ; genetics ; Adult ; DNA Mutational Analysis ; DNA, Neoplasm ; genetics ; Exons ; Family ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Pheochromocytoma ; genetics ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

OBJECTIVETo study the cytotoxic T lymphocyte (CTL) response induced by dendritic cells (DC) transduced with recombinant adenovirus vector bearing hepatitis B virus surface antigen (HBsAg) gene in hepatocellular carcinoma HepG2. 2. 15 cells in vitro.

METHODSFull length HBsAg cDNAs were subcloned into pIND vector, followed by being cloned into pShuttle vector. The HBsAg gene fragments resulted from the pShuttle-S digested with PI-Sce and I-Ceu were linked to the linear adeno-X virus DNA. After packaged with HEK293 cells, the adenovirus expression vector was obtained. Then the recombinant adenovirus expression plasmid AdVHBsAg was transfected into human monocyte-derived dendritic cells, to construct AdVHBsAg hepatocarcinoma tumor vaccine. The effectiveness of transfection was detected by Western blot. Surface molecules of AdVHBsAg-DC were detected by FACS. Autologous T cell proliferation stimulated by AdVHBsAg-DC was detected by 3H-TdR assay. Cytotoxic CTL activity induced by AdVHBsAg-DC in vitro was detected by LDH assay.

RESULTSHBsAg gene in the inserted DNA of AdVHBsAg was confirmed by PCR, and predictive fragments proved by restriction enzyme digestion analysis were exhibited. Cell pathological changes appear after 10 days HEK293 cells transfected AdVHBsAg. Western blot analysis showed that HBV surface antigen gene was expressed in transfected DC, indicating that the transfection was effective. AdVHBsAg-DC was able to upregulate CD1a, CD11c, CD80, CD86 and HLA-DR. Autologus T cell proliferation induced by AdVHBsAg-DCs was significantly higher than that in DC control group and LacZ-DC group (P < 0.05). AdVHBsAg-DC activated CTL presented the specific killer ability to the hepatocellular carcinoma cells expressing HBsAg.

CONCLUSIONDC transduced with recombinant adenovirus HBsAg can express HBV-related hepatocellular carcinoma antigen (HBsAg), and AdVHBsAg-DC can induce potent immune response against HBsAg-positive hepatocellular carcinoma cells in vitro.

Adenoviridae ; genetics ; Antigens, CD1 ; metabolism ; CD11c Antigen ; metabolism ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; virology ; Cell Line, Tumor ; Cell Proliferation ; Cytotoxicity, Immunologic ; immunology ; Dendritic Cells ; cytology ; immunology ; metabolism ; Genetic Vectors ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Humans ; Liver Neoplasms ; immunology ; pathology ; virology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Transfection

OBJECTIVETo investigate K(ATP) channel function of cardiomyocytes isolated from the left ventricular wall of rats with or without abdominal aortic constriction at different time points under normal or simulated ischemic conditions.

METHODSMale Wistar rats were randomized into 4 groups (n = 10 - 13): 4-week sham-operated group (F4), 4-week aortic-banded group (T4), 12-week sham-operated group (F12), 12-week aortic-banded group (T12). Chronic pressure overload model was established by abdominal aortic constriction. Left ventricular myocytes were isolated by modified Langendorff perfusion method post in vivo hemodynamical measurements. The whole-cell patch-clamp technique was used to record transient outward current of K(ATP) channel on myocytes under normal and simulated ischemic perfusion conditions. The current densities of K(ATP) channel between F4 and T4 group, F12 and T12 group were compared under 0 mV of test potential.

RESULTSSBP, DBP and MBP were significantly increased in T4 group compared to F4 group, but were similar between T12 and F12 groups. LVEDP and +/- dp/dtmax were similar between T4 and F4 groups and LVEDP was significantly increased while +/- dp/dtmax significantly reduced in T12 group than that in F12 group. Whole-cell membrane current densities were similar between F4 and T4 group or F12 and T12 group under normoxic condition, the K(ATP) current densities increased dramatically in T12 group [(28.11 +/- 3.91) pA/pF vs (11.55 +/- 1.17) pA/pF, P < 0.01], but not in T4 group [(14.09 +/- 5.74) pA/pF vs (11.74 +/- 3.68) pA/pF, P > 0.05] in myocytes exposed to ischemic solution for 25 minutes. The total number of K(ATP) channel in ventricular myocytes was similar between F4 and T4 group or F12 and T12 group.

CONCLUSIONSThe sarcolemmal K(ATP) channel was more sensitive to ischemia and the current magnitude was significantly increased at the stage of congestive heart failure. The functional change of K(ATP) channel occurred before the increase of total number of K(ATP) channel.

Animals ; Disease Progression ; Heart Failure ; metabolism ; physiopathology ; KATP Channels ; metabolism ; Male ; Myocytes, Cardiac ; pathology ; Patch-Clamp Techniques ; Rats ; Rats, Wistar

OBJECTIVEInadequate postnatal nutritional support is an important factor contributing to growth failure, which leads to poor neurological outcome. In this study, co-word analysis was used to investigate the research on nutrition of premature infants over the last six years in China, describe the research trend in this field in China, and provide possible directions for future research.

METHODSA literature search was performed in January 2013 using the CNKI database and the key words "preterm infant" and "nutrition". A total of 772 articles were retrieved. Then high-frequency key words were extracted using Excel 2010 to create a co-occurrence matrix. Finally, a visualized network was built using Ucinet 6.0.

RESULTSThe knowledge domain map of research on nutritional support for premature infants in China showed that the major topic of relevant research is still the combination of parenteral nutrition and enteral nutrition, with the goal of maintaining appropriate growth rates in premature infants. Researchers have paid much attention to the adverse effects of parenteral nutrition. Feeding intolerance is still the main problem in nutritional support, especially enteral nutrition, for premature infants.

CONCLUSIONSA visualized network of current research on nutrition of premature infants in China has been created, and a knowledge domain map has been drawn to reflect the hot topics in this field of study over the last six years.

Enteral Nutrition ; Humans ; Infant Nutritional Physiological Phenomena ; Infant, Newborn ; Infant, Premature ; physiology ; Parenteral Nutrition

It's known to all that the refractory ascites treatment has so far been a very difficult clinical problem. We have extracted much experience from the practical techniques used in the refractory ascites treatments of more than 1,000 cases, and have developed the ascites ultrafiltration & concentration therapeutic instrument--FSCLZLY-A. The clinical applications show that it is very effective. Its effective rate is about 72.08%. Therefore, it is a very useful and important medical device for refractory ascites, for the improvement of renal function, and for the prevention of the infection of abdominal cavity.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Ascites ; therapy ; Equipment Design ; Female ; Humans ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Treatment Outcome ; Ultrafiltration ; instrumentation ; methods

G-quadruplexes with different conformations often exhibit different binding abilities to proteins and play different physiological functions. It is of great significance to analyze the factors affecting the configuration of G-quadruplex to explore its physiological function. Different from DNA, RNA sequences have shown parallel G-quadruplex structures in previous reports. However, we found that the only RNA sequence, the 22-nt telomere RNA fragment (ORN-N, A