AIM: To explore the frequency of drug injection of alloxan diabetes on the established model of rabbit.METHODS: Thirty-six healthy rabbits, weighing 2-2.5kg, were randomly divided into one time drug injection group (group A, n=12), two times drug injection group (group B, n=12) and three times drug injection group (group C, n=12). Each rabbit was injected with a total amount of 150mg/kg of alloxan. Fasting blood glucose was measured. The success rate and death rate of each group were also calculated.RESULTS: The success rate of diabetic rabbit model in group B was higher than that in group A (P<0.01) and its death rate was lower than that of group A (P<0.01); the success rate of diabetic rabbit model in group C was highest and the death rate was the lowest in three groups(P<0.01). CONCLUSION: Multiple administration of alloxan can improve success rate in establishing diabetic rabbit model with decreased death rate and increased stability.

BACKGROUND: The treatment of degenerative knee osteoarthritis is mainly to relieve pain, restore knee function, improve quality of life, delay knee replacement, and reduce the number of revisions. Knee replacement is currently the most common treatment for this disease, but it costs much, has great trauma and high risk, often results in prosthesis loosening and peripheral infection, and has many adverse reactions. OBJECTIVE: We hypothesize that knee-preserving arthroscopic debridement for treatment of knee degenerative osteoarthritis in the elderly patients costs less, is effective, safe, and reliable. METHODS: A total of 212 elderly patients (knees) with degenerative knee osteoarthritis who receive treatment in the Second Hospital of Chaoyang (Liaoning Province, China) will be included in this study. These patients will be assigned to two groups according to patient's conditions and wishes (n = 106/group). In the control group, intra-articular injection of sodium hyaluronate will be performed, followed by oral administration of non-steroidal anti-inflammatory drugs, conventional physiotherapy, and quadriceps functional exercise. In the arthroscopic debridement group, arthroscopic debridement will be performed followed by oral administration of non-steroidal anti-inflammatory drugs, conventional physiotherapy, and quadriceps functional exercise. All patients will be followed up for 1 week, 1 month, 3 months, 6 months, 1 year, and 2 years. RESULTS AND CONCLUSION: The primary outcome measure is the percentage of the number of patients with Hospital for Special Surgery (HSS) knee score ≥ 85 points at 2 years after surgery, which will be used to evaluate knee function recovery. The secondary outcome measures are the percentage of the number of patients with HSS knee score ≥ 85 points before surgery, 1 week, 1 month, 3 months, 6 months, and 1 year after surgery; HSS score, Visual Analogue Scale (VAS) score, The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), knee range of motion, hospitalization costs, and knee X-ray morphology before surgery, 1 week, 1 month, 3 months, 6 months, 1 year, and 2 years after surgery, medical costs after 2 years of treatment, incidence of adverse reactions at 1 week, 1 month, 3 months, 6 months, 1 year and 2 years after surgery. Findings from this study will reveal whether arthroscopic debridement for the treatment of degenerative knee osteoarthritis in the elderly patients has advantages of less adverse reactions, low treatment costs, and can effectively restore knee function. This trial has been approved by the Second Hospital of Chaoyang, Liaoning Province, China (approval number:2017-08-01).All protocols will be in accordance with Declaration of Helsinki,formulated by the World Medical Association.Written informed consent will be provided by participants. This trial was designed in June 2017. The recruitment of subjects and data collection will begin in June 2018. The recruitment of subjects will be finished in December 2018. Outcome measures will be analyzed in June 2021. This trial will be completed in August 2021. The results of the trial will be reported in a scientific conference or disseminated in a peer-reviewed journal. This trial had been registered in the Chinese Clinical Trial Registry (registration number: ChiCTR1800015208). The version of this study protocol is (1.0).

The aim of this study was to identify the relationship between susceptibility of children to acquired aplastic anemia (AA) and HLA-A, -B, -DRB1 alleles. 80 children with AA were enrolled in this study. Among of them, 34 patients collected from tissue typing test centers of Nanfang Hospital; 46 patients were diagnosed at Department of Pediatrics of Sun Yat-Sen Memorial Hospital. In these patients, 48 were males, 32 were females, and with average age 8.1 years old, 6 cases were non-severe AA (nSAA), 74 case were severe AA (SAA). The healthy control group consisted of 109 donors who were from the same area. All the patients and healthy controls were of Han Chinese, and all were unrelated individuals. The polymerase chain reaction sequence specific primers (PCR-SSP) was used to analyze the polymorphism of HLA-A, -B and -DRB1 alleles. Pearson Chi-square or continuity correction or two-sided Fisher's exact test were used. The results showed that the genotype frequency of HLA-B*48:01 and DRB1*09:01 were significantly higher in children with AA as compared with healthy controls (P < 0.05). The genotype frequency of HLA-B*51:01, DRB1*03:01 and DRB1*11:01 were significantly lower in children with AA as compared with healthy controls (P < 0.05). Besides, the results also demonstrated that the genotype frequencies of HLA-B*48:01 and DRB1*09:01 were significantly higher in SAA as compared with controls, the genotype frequencies of B*51:01, DRB1*03:01 and DRB1*11:01 were significantly lower in SAA, as compared with controls. In conclusion, HLA-B*48:01 and DRB1*09:01 are related with children AA, and may be susceptible alleles to the development of children AA. Besides, the expression of HLA-B*51:01, DRB1*03:01 and DRB1*11:01 are low in children with AA, whether they are relative protection alleles of children needs to be further studied.
Adolescent ; Alleles ; Anemia, Aplastic ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DRB1 Chains ; genetics ; Humans ; Infant ; Male ; Polymorphism, Genetic

OBJECTIVETo verify the efficacy of Jianpi-xingniao needling therapy on prevention and treatment of motion sickness.

METHODSSixty volunteers of motion sickness were randomized into an acupuncture group and a delayed acupuncture group, 30 cases in each one. In the acupuncture group, acupuncture was given at Baihui (GV 20), Sishencong (EX-HN 1), Neiguan (PC 6), Zusanli (ST 36) and Hegu (LI 4). The needles were retained for 20 min. The treatment was given twice a week and 10 treatments were required. In the delayed acupuncture group, acupuncture was postponed, meaning no acupuncture during observation stage. Graybel scale was adopted to observe the score of symptoms and physical signs of the subjects of motion sickness before and after intervention. The efficacy was compared between the two groups.

RESULTSTwenty-five cases in the acupuncture group and 22 cases in the delayed acupuncture group were included in the statistical analysis. The score of symptoms and physical signs of motion sickness was reduced significantly after treatment in the acupuncture group as compared with that before treatment (10.12 +/- 3.37 vs 0.92 +/- 0.40, P < 0.05). The score in the acupuncture group was lower apparently than that in the delayed acupuncture group (0.92 +/- 0.40 vs 9.86 +/- 2.53, P < 0.05). The difference was not significant before and after treatment in the self-comparison of the delayed acupuncture group (P > 0.05). The total effective rate was 96.0% (24/25) in the acupuncture group, which was significantly better than 0.0% (0/22) in the delayed acupuncture group (P < 0.01).

CONCLUSIONJianpixingniao needling therapy relieves the symptoms of motion sickness in the patients and achieves a better clinical efficacy.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Motion Sickness ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo establish chemical pattern recognition method for the identification and evaluation of Atractylodes macrocephala.

METHODThe chemical constituents in methanol extract of 32 samples of A. macrocephala were determined by HPLC. The fingerprints were obtained and were handled by hierarchical clustering analysis and stepwise discriminant analysis.

RESULT AND CONCLUSIONAccording to the result of classification, all samples collected were devided into three Grades--the superior, the ordinary and the fake. Chemical pattern recognition method was established. It may be of practical value for the quality control of A. macrocephala.

Atractylodes ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Pattern Recognition, Automated ; Plants, Medicinal ; chemistry ; Quality Control ; Rhizome ; chemistry

OBJECTIVETo prepare and purify recombinant human NAMPT and NAMPT (H247A) proteins and to detect their enzymatic activity.

METHODSUsing pcDNA3.1-hnampt as template, full-length hnampt was sub-cloned into pET-11a(+) plasmid. The hnampt (H247A) mutant was obtained by site-directed mutagenesis. The plasmids were introduced in Escherichia coli BL21star for protein expression. The recombined NAMPT and NAMPT (H247A) proteins were purified by flowing through nickel column and size-exclusion column. The target proteins were confirmed by SDS-PAGE and mass spectrometry detection. The enzymatic activities of recombinant proteins were assessed by solution NMR.

RESULTThe DNA sequences showed that hnampt (wild type) and hnampt (H247A) (mutation) were cloned into pET-11a(+). The recombinant proteins were expressed in Escherichia coli BL21star in soluble form. The purified protein was confirmed to be NAMPT with a molecular weight of 56 KD. The enzyme activity of NAMPT (H247A) was dramatically decreased compared to wild-type NAMPT.

CONCLUSIONThe recombinant hNAMPT and hNAMPT (H247A) proteins have been successful prepared and purified. The H247A mutation dramatically decreases the enzymatic activity of NAMPT.

Base Sequence ; Cytokines ; genetics ; isolation & purification ; metabolism ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Nicotinamide Phosphoribosyltransferase ; genetics ; isolation & purification ; metabolism ; Plasmids ; genetics ; Recombinant Proteins ; genetics ; metabolism ; Transformation, Bacterial

This study was aimed to explore the frequency of PTPN11 mutation in children with leukemia and its clinical significance. Genomic DNAs were extracted from peripheral leukocytes of 131 patients with leukemia, including 101 cases of ALL, 26 cases of AML, 3 cases of CML and 1 case of juvenil myelomonocytic leukemia (JMML). The sequences of PTPN11 exons 3, 8, 13 were amplified by polymerase chain reaction (PCR), and the clinical characteristics of positive patients were analyzed. The results indicated that the PTPN11 mutation was found in 10 cases (9.9%) from newly diagnosed 101 cases of ALL. Grouping the newly diagnosed ALL children by various clinical features, it was found that the PTPN11 mutation did not show associations with sex, age, white blood cell (WBC) count, prednisone test sensitivity, clinical risk and disease recurrences at the first visit (P > 0.05). PTPN11 mutations were found in 2 cases out of 26 AML patients, including one AML-M(2) and one AML-M(4). No PTPN11 mutation in 3 CML patients was found. Exon 13 mutation of PTPN11 gene was found in 1 case of JMML. It is concluded that the E76 of exon 3 is the hot spot of PTPN11 mutation in children leukemia. The novel G503E (1508G > A) mutation is detected in one JMML patient. The PTPN11 mutation does not associate with the sex, age, WBC count, prednisone sensitive test and early recurrence.
Adolescent ; Base Sequence ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukemia ; genetics ; Male ; Mutation ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; genetics

Objective To evaluate the diagnostic value of brain SPECT imaging with a novel Aβ plaque probe,131 I-2-(4'-dimethylaminophenyl) -6-iodoimidazo[ 1,2-α ] pyridine ( 131 I-IMPY) in early AD.Methods Thirteen patients with AD (3 males,10 females,age ranged 52 - 79 y),11 with mild cognitive impairment (MCI,4 males,7 females,age ranged 48 - 67 y) and 14 normal controls (6 males,8 females,age ranged 42 - 67 y) were enrolled in this study.131I-IMPY SPECT imaging was acquired in 2 -3 h after the agent injection.ROIs were drawn on cerebral lobes and cerebellum.The ratios of mean radioactivity of cerebral lobes over cerebellum (Rcl/cb) were calculated.The t-test was used for data analysis.Results In patients with MCI,Rcl/cb ratios were increased in parietal gyrus,temporal gyrus and frontal gyrus (right:1.15±0.18,1.18±0.12,1.14±0.14; left:1.16±0.11,1.19±0.18,1.15±0.09)compared with those in normal control group ( right:1.02 ± 0.12,1.05 ± 0.14,1.01 ± 0.12 ; left:1.03 ±0.13,1.05 ±0.13,1.01 ±0.14; t:2.1642 to 2.8757,all P ＜0.05).Rcl/cb ratios of basel ganglia and occipital gyms in MCI group (right:0.92 ±0.18,1.12 ±0.15; left:0.94 ±0.15,1.13 ±0.17) showed no statistical difference compared with those in normal control group (right:0.82 ±0.15,1.06 ±0.18;left:0.85 ±0.16,1.08 ±0.15; t:0.7805 to 1.4344,all P＞0.05).In patients with AD,Rcl/cb ratios were increased in parietal,temporal,basal ganglia and occipital lobes (right:1.16 ±0.19,1.24 ±0.17,1.16 ±0.13,1.14±0.11,1.23±0.10; left:1.17±0.21,1.25±0.15,1.18±0.08,1.17±0.16,1.25±0.11)compared with those in normal control group( t:2.1001 to 6.2789,all P ＜0.05).Rcl/cb ratios of parietal,temporal and frontal lobes in AD group showed no statistical difference compared with those in MCI group (t:0.1316 to 0.9806,all P ＞ 0.05 ),while Rcl/cb ratios of basal ganglia and occipital lobes in AD group were increased compared with those in MCI group ( t:2.0850 to 3.6772,all P ＜ 0.05 ).Conclusion 131 I-IMPY as a β- amyloid plaque probe for brain SPECT imaging may be potentially helpful for early diagnosis of AD.

Objective To explore the biological behavior and pathologic mechanism of ectopic decidua hemorrhage or acute postpartum hemorrhagic deciduosis caused by typical decidua tissue outside the uterine cavity under the influence of ovarian and placental hormones,and provide reference for accurate diagnosis and effective treatment. Methods From August 2017 to January 2018,there were 461 term-pregnant women undergoing ceasarian section in maternal and child health hospital of Qingbaijiang district,of whom 3 cases with ectopic deciduas were diagnosed.The clinical characteristics and microscopic features of 3 cases were retrospectively analyzed.The risk assessment to human health as well as effective control measures of ectopic deciduas were further elucidated along with the relevant references. Results Three patients with ectopic decidua generally had no obvious clinical symptoms or endometriosis,though dysmenorrheal and infertility might happen before pregnancy.Only an intraoperative incidentally finding of specific lesions varied from vacuolar plaques, hyperemia,vascular nodules, solid nodules and local hemorrhagic change.Under light microscope, the decidualized stroma revealed large polygonal cell aggregates, without nu-clear atypia.Due to the clinical features of abnormal angiogenesis and inflammatory reactions, ectopic decidua may cause severe cases such as acute postpartum hemorrhagic deciduosis,which could provide a basis for clinical comprehensive understanding and scientific prevention and treatment.Conclusion Ectopic decidua can happen in uterine seromuscular layers,ovary,great epiploon,cervix,vagina and so on,which is easy to be misdiagnosed or fails to be examined due to asymptom and nonspecific physical signs.It hasn’t been unequivocally clarified and widely recognized,nor earned extensively considered for rarely life-threatening haemorrhagic deciduosis in the prepartum and postpartum peri-od,which must be taken effective first-aid measures such as surgical intervention to protect maternal-fetal health.

OBJECTIVETo evaluate the role of leptin in neointimal formation and related mechanisms.

METHODSFemoral arterial injury was induced in wild-type (Wt, n = 10), leptin-deficient (Lep(-)/-, n = 12), and leptin receptor-deficient (LepR(-)/-, n = 10) mice. Leptin treatment studies (tail vein injection of adenovirus expressing murine leptin on the RSV promoter, ad-leptin) were performed on Lep(-)/- (n = 5) and LepR(-)/- (n = 4) mice. Intimal (I) and medial (M) areas were measured and the ratio of I/M was calculated. Smooth muscle cells were detected by smooth muscle alpha-actin staining using an alpha-actin monoclonal antibody. Cellular proliferation was analyzed with BrdU Staining Kit and the number of BrdU-positive cells was counted manually. Plasma leptin level was measured by ELISA.

RESULTSThe I/M ratio of Lep(-)/- and LepR(-)/- mice was significantly lower than that in Wt separately (Lep(-)/- vs. Wt = 0.80 +/- 0.14 vs. 1.50 +/- 0.22, P < 0.01; LepR(-)/- vs. Wt = 0.55 +/- 0.20 vs. 1.50 +/- 0.22, P < 0.05). Plasma leptin level was significantly increased in Lep(-)/- and LepR(-)/- mice post leptin treatment. I/M was significantly increased in Lep(-)/- mice receiving ad-leptin compared with untreated Lep(-)/- mice (P < 0.05), while I/M was similar between LepR(-)/- mice with and without ad-leptin treatment (P > 0.05). The changes on number of positive alpha-actin and BrdU stained smooth muscle cells were consistent with the neointimal formation findings in various groups.

CONCLUSIONSMice lacking leptin or the leptin receptor were protected from neointimal formation following vascular injury. Leptin treatment increased neointimal formation in Lep(-)/- but not in LepR(-)/- mice, suggesting leptin receptor activation and vascular smooth muscle cell proliferation played a pivotal role on neointimal formation post-injury in this model, giving an evidence that high plasma leptin level is a risk factor for neointimal formation.

Actins ; analysis ; Animals ; Cell Proliferation ; Leptin ; blood ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Smooth, Vascular ; pathology ; Receptors, Leptin ; metabolism ; Tunica Intima ; pathology