BACKGROUND:Transgenosis of basic fibroblast growth factor (bFGF) gene has been successfully performed into the muscle satellite cells of rat extraocular muscles in the previous study of the research group, proving that bFGF could express in the myoblasts of extraocular muscles, also promote cell proliferation and differentiation.OBJECTIVE: To further investigate the methods for regulating the expression of the bFGF in myoblasts following transfection. METHODS: Target gene bFGF was connected with inducing expression vector pcDNA4/T0/myc-His?A, followed by masculine clone sequencing identified by colony PCR and enzyme digestion, EcoR I and Hind III restriction enzyme digestion, as well as Xho I single enzyme verification. C2C12 myoblasts antibiotics sensitivity was screened and finally defined. By use of lipofection transfection technology, cell lines where C2C12 stably expressed pcDNA6/TR were estabolishd and then identified by Western blot. The pcDNA4/TO/myc-His?A-bFGF was transfected into pcDNA6/TR- C2C12 cells. The bFGF expression and secretion in C2C12 cells following tetracycline-induced pcDNA4/TO/myc-His?A-bFGF transfection were determined by immunofluorescence and Western blot, the controls were established.RESULTS AND CONCLUSION: ① The conjunction between the bFGF and inducing expression vector pcDNA4/TO/myc-His?A was proved successfully by sequencing comparison, double digestion and single digestion. ②The minimal lethal concentration of blasticidin to C2C12 cells was 10 mg/L, while that of zeocin was 750 mg/L. ③ The pcDNA6/TR-C2C12 cell lines were established correctly. ④ The myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF were positive for gene expression, those untreated exhibited a negativity; bFGF protein could be produced in myoblasts treated by tetracycline and transfected with pcDNA4/TO/myc-His?A-bFGF, the production reached a peak at 24 hours, while those untreated can not produce bFGF protein. Results suggest that the bFGF expression in the myoblasts can be controlled by tetracycline inhibition and regulatory systems.

Objective CD8 +T cells increased in the airway of patients with chronic obstructive pulmonary disease and exis -ted constantly .The aim was to investigate the role of CD 8 +T-cells in rats with chronic bronchitis ( CB) which was induced by cigarette smoking and intratracheal injection with lipopolysaccharide ( LPS) . Methods 18 health Wistar rats were radomly divided into sham smoking group(group A), CB group(group B) and N-acetylcysteine prevention group (group C).The rats in group B and group C re-ceived intratracheal injection with LPS twice and exposed to cigarette smoking for 4 weeks to induce CB model .The rats in Group C re-ceiving intragastric administration with N-acetylcysteine (NAC)(200mg/kg) before received LPS and smoking.Group A was the sham smoking group.The lung tissue of all rats were stained by HE then evaluated about pathological scores .The expression of nuclear fac-tor-κB (NF-κB), major histocompatibility complex class I (MHCI), CD8 +T cell and Vascular endothelial growth factor (VEGF) in airway were detected by immunohistochemisty which was stained by labeled streptavidin biotin method . Results The pathological scores of airway ( 10 .83 ±3 .31 ) in group B were higher than (1.17 ±2.40) in group A(P <0.05).The pathological scores of airway(4.66 ±2.25) in group C were less than (10.83 ±3.31) in group B(P <0.05).The expression of NF-κB(4.84), MHC I (2.48),CD8 +T cell(5.35)and VEGF(5.02) in airway increased in group B when compared with (1.18, 1.25, 1.33) and (1.18) in group A respectively(P <0.05).The expression of NF-κB (2.18), MHC I(1.46),CD8 +(2.35)and VEGF(2.02) in airway decreased in group C when compared with (4.84), MHC I(2.48),CD8 +T cell(5.35)and VEGF(5.02) in group B respectively (P<0.05 ). There were positive correlations between the expression of NF-κB, MHC I and CD8 +T cells in airways(r=0.670, r=0.701, respec-tively, all P<0.01).There were positive correlations between the expression of CD 8 +T cells and VEGF the pathological scores of air-ways(r=0.689, r=0.782, respectively, all P<0.01). Conclusion NAC can inhibit airway inflammation which is regulated by CD8 +T-cells and VEGF through suppressing the expression of NF -κB and MHC I.

Objective To investigate the applicability of clinical grade and a non-contact measurement method in evaluation of underneath eye wrinkles and to compare two methods.Methods A lot of 46 healthy Chinese women were recruited for this study.Underneath eye wrinkles severity was evaluated using clinical grade and a non-contact measurement method.The correlations were calculated for clinical grade and non-contact measurement parameters and age.The non-contact measurement parameters were classified by factor analysis.The correlations between age,clinical grade and factors were analyzed.Results The correlation coefficient between clinical grade in comparison to subject's age was 0.818.The parameters getting from non contact measurement were obviously correlated with age and clinical grade except SEr and SEsc;the correlation coefficients between parameters and age were-0.601 to 0.605;the correlation cofficients between parameters and clinical grade were-0.630 to 0.570.The non-contact measurement parameters could be classified into two factors;one represented wrinkle depth and roughness;the other represented wrinkle width and counts.These two factors were also obviously correlated with age and clinical grade.Conclusions Clinical grade and non contact measurement methods are both applicable in evaluation of underneath eye wrinkles.The parameters getting from two methods are obviously correlated with each other.

Objective: To investigate the diagnostic value of serum levels of aquaporin-4 (AQP4) and matrix metalloproteinase-9 (MMP-9) in predicting malignant cerebral infarction in patients with cerebral infarction. Methods: The clinical data about 66 patients with hemisphere massive cerebral infarction were collected. According to clinical symptoms and brain computed tomography (CT) findings, the patients were divided into malignant cerebral infarction group (MCI) and non-MCI group. The serum levels of AQP4 and MMP-9 were detected 1, 4, 7 days. The diagnostic value of AQP4 and MMP-9 in predicting occurrence of MCI was analysed by receiver operating characteristic (ROC) curve. Results: The serum levels of AQP4 and MMP-9 in MCI group were significantly higher than those in non-MCI group in the fourth day (P<0.05). In MCI group, AQP4 and MMP-9 in the fourth day were significant higher than other two time points (P<0.05). But in non-MCI group, AQP4 and MMP-9 in different time points have no differences (P>0.05). ROC curve analysis showed that the aera under curve for the serum levels of AQP4 was 0.654 (95% CI: 0.516-0.792). When the serum AQP4=23.49 μg/L, the sensitivity and specificity were 0.57, 0.74, respectively. Conclusions Serum AQP4 and MMP-9 levels were increased in patients with MCI. Combined detection of serum AQP4 and MMP-9 is helpful for early differential diagnosis of MCI.

The clinical characteristics and gene mutation characteristics of a child with typical maple syrup urine disease were analyzed retrospectively.The child is a boy, who showed unexplained milk refusal, poor reaction, foaming at the mouth, and encephalopathy symptoms 7 days after birth.The total leucine concentration was abnormally increased by blood tandem mass spectrometry, and the results of urine gas chromatography/mass spectrometry suggested that the concentrations of 2-hydroxy isovaleric acid, 2-keto isovaleric acid, 2-keto-3-methylpentanoic acid and 2-keto-isohexanoic acid were significantly increased.The gene detection results showed that c. 1028delC (p.S343Lfs*9) homozygous mutation was found in the BCKDHB gene.Understanding the clinical symptoms and gene mutation characteristics of this disease can help with the early detection and early diagnosis of this disease, so as to improve its prognosis to the greatest extent.

Objective:To explore the effects of nutrition guidance combined with exercise intervention among elderly patients with sarcopenic obesity in community.Methods:From January 2014 to November 2015, we selected 120 elderly patients with sarcopenic obesity of Xixi Street Community Health Service Center in Hangzhou as subjects by convenience sampling. All of patients were divided into control group and intervention group according to the odd and even of visit number, 60 cases in each group. Control group carried out the routine health education, while intervention group implemented the balanced diet combined with exercise intervention as well as increase whey protein appropriately. Six months after intervention, we compared the body mass index (BMI) , grip, calf girth, leg speed, 5 times sitting up test, human muscle percentage and body fat percentage of patients between two groups.Results:Before intervention, there were no statistical differences in the indexes of patients between two groups ( P>0.05) . Six months after intervention, the BMI of intervention group and control group was (25.39±0.95) kg/m 2 and (25.46±0.93) kg/m 2 respectively with no statistical difference ( t=0.36, P>0.05) . The six indexes, grip, calf girth, leg speed, 5 times sitting up test, human muscle percentage and body fat percentage, of intervention group were (23.43±2.04) kg, (32.60±1.34) cm, (0.83±0.10) m/s, (11.05±1.50) s, (47.69±2.87) % and (33.68±2.20) % respectively, all better than those of control group with statistical differences ( t=-4.62, -2.06, -2.62, 2.46, -3.68, 2.46; P<0.05) . Conclusions:Nutrition guidance combined with exercise intervention can improve the functional indexes of elderly patients with sarcopenic obesity in community, such as muscle strength, and delay the functional decline caused by sarcopenia.

Prosthetic joint infection (PJI) is a catastrophic complication after joint replacement.It has become the primary cause of revision surgery after knee arthroplasty and the third cause of revision after hip replacement surgery.The risk factors involve the patient (intrinsic factor) and the environment (extrinsic factor),which can be further divided into uncontrollable factors such as the history of surgical infection and other factors related to age and sex of tumor,as well as the risk factors caused by patients' own characteristics and associated diseases,such as body mass index,smoking,diabetes,rheumatoid arthritis and medicine use.The authors summarize the patients' own characteristics,associated diseases and medication to comprehensively evaluate and understand how to reduce the risk of PJI in the perioperative period,so as to provide some reference for clinical treatment.

Objective To investigate the natural evolution of postoperative distal adding-on in Lenke 1A and 2A adoles-cent idiopathic scoliosis(AIS)patients,and to explore the risk factors for the progression of distal adding-on.Methods From Ju-ly 2006 to July 2012,a total of 197 AIS patients with Lenke 1A or 2A curves underwent posterior selective thoracic instrumenta-tion and fusion surgery.Among which,44 patients(22.3%)with postoperative distal adding-on were recruited in this study.There were 39 female and 5 male,with an average age of(15.0±2.1)years.The mean Cobb angle of main thoracic curve was 49.3°±9.3°. The first postoperative radiograph indicating distal adding-on and the last follow-up radiograph were compared:make the measure-ment of the disc angle below lowest instrumented vertebra(LIV),and the distance between the vertebra below LIV(LIV+1)and cen-tral sacral vertical line(CSVL).Distal adding-on could be classified into progressive group and non-progressive group according to its natural evolution during follow-up.If the disc angle increased> 5°or the LIV+1-CSVL distance increased>5 mm,the pa-tients were assigned into progressive group; Otherwise, the patients were assigned into non-progressive group. Using Student T test, χ2test or Fisher exact test, the predicted risk factors for progression were screened for further Logistic regression. Results Among the 44 patients enrolled in the study,17 patients(38.6%)had progressive adding-on while 27 patients(61.4%)had non-progressive adding-on.The Risser sign was significantly lower in progressive group than non-progressive group(t=4.399,P<0.001). Besides,more patients had LIV proximal to substantially stable vertebra(SSV)in progressive group than non-progressive group (Fisher exact test value=18.142,P<0.001).The improvement of shoulder imbalance was significantly better in progressive group than non-progressive group(t=3.011, P=0.002). According to Logistic regression, the low Risser sign and LIV proximal to SSV were independent risk factors for progression of distal adding-on.Moreover,the self-image domain of SRS-22 Scores was remark-ably lower in progressive group than non-progressive group(t=2.321,P=0.014).Conclusion Distal adding-on could be classi-fied into progressive group(40%)and non-progressive group(60%)according to its natural evolution.The risk factor for its progres-sion included skeletal immaturity and LIV proximal to SSV.Moreover,the progression of distal adding-on might compensate for the shoulder imbalance during follow-up.

Objective:To construct a column-line diagram diagnostic model based on serum and joint fluid inflammatory markers for the diagnosis of periprosthetic joint infections (PJI) after joint arthroplasty and to validate its predictive ability.Methods:The clinical data of 181 patients diagnosed with PJI or aseptic loosening in the Department of Orthopedics of the First Affiliated Hospital of Chongqing Medical University from January 2015 to June 2020 were retrospectively collected as a modeling group. The best indicators for diagnosing PJI were screened by lasso regression, single-factor and multifactor analysis. By comprehensively considering the weights and intrinsic connections of the indicators, a column-line diagram diagnostic model was constructed and used to develop a clinical decision support system (CDSS). Prospectively, the clinical data of patients diagnosed with PJI or aseptic loosening in the Department of Orthopedics of the First Hospital of Chongqing Medical University from July 2020 to December 2022 were collected as a validation group, and the diagnostic performance of the column-line diagram model was externally validated by methods such as receiver operating characteristic curve (ROC).Results:There were 85 cases of PJI in the 181 cases modeling group and 23 cases of PJI in the 49 cases validation group. Among the 27 potential factors analyzed by lasso regression analysis, body mass index (BMI), blood tests including platelet (PLT), absolute lymphocyte value, interferon γ (IFN-γ), ESR, IL-6, C-reactive protein, D-dimer, and joint fluid tests including C-reactive protein, IL-1, IL-4, IL-6, percentage of multinucleated neutrophils (PMN%), and CD64 may be potential indicators for the diagnosis of PJI. Univariate found significant differences between hematologic tests including sedimentation, C-reactive protein, IL-6, D-dimer and joint fluid tests including C-reactive protein, joint fluid CD64 index, C-reactive protein, IL-1, IL-4, IL-6, PMN%( P<0.05). Further multifactorial regression analysis screened serum IL-6, D-dimer, joint fluid CD64 index, C-reactive protein, IL-1, IL-4, IL-6, and percentage of multinucleated neutrophils, and based on that, the column-line graph model and CDSS system were constructed. The area under the ROC in the validation group was 0.978, and the AUC in the internal validation was 0.995; the C-index of the calibration curve was 99.50%, and the C-index of the internal validation was 99.53%, suggesting that the column-line diagram model has a good predictive ability. Conclusions:The column-line diagram for diagnosing PJI based on multiple diagnostic indicators showed good diagnostic performance. The CDSS system constructed by column-line diagrams could assist clinicians in diagnosing PJI and making reasonable strategies in time.

Objective:To explore the correlation between structural chromosomal abnormality and clinical characteristics of a child featuring gonadal dysplasia.Methods:A 13-year-old child who was admitted to Lianyungang Maternal and Child Health Care Hospital on February 7, 2023 for primary amenorrhoea and occasional abdominal pain was selected as the study subject. Clinical data of the child was collected, and peripheral blood samples of the child and her parents were collected. G-banding chromosomal karyotyping and copy number variation sequencing (CNV-seq) were carried out. "Pseudodual centromere isochromosome X" and "psu idic(X)" were used as keywords to search the CNKI, Wanfang and PubMed databases, and the search period was set as from January 1, 2002 to June 1, 2023. Relevant literature on the structural abnormality of X chromosome was searched and analyzed retrospectively.Results:The child has a height of 153 cm and weighed 45 kg. She has no obvious facial dysmorphism. Laboratory tests showed that she had higher FSH and luteinizing hormone, and lower E2. Ultrasonography showed that she had small ovaries and rudimentary uterus. She was found to have a karyotype of 46, X, psu idic(X)(q21.3)[40]/mos 45, X[3], whilst both of her parents had a normal karyotype. CNV-seq showed that she had a 63.27 Mb deletion in Xq21.32q28 and a 91.59 Mb duplication in Xp22.33q21.32 (mosaicism rate = 74%). A total of 11 relevant literature were retrieved. Clinical phenotypes of patients with similar structural chromosomal abnormalities were diverse, which was closely related to the mosaicism rate of the 45, X karyotype and the location of the breaking point.Conclusion:46, X, psu idic(X)(q21.3)/45, X probably underlay the dysplasia of uterus and ovary and sex hormone abnormalities in this child, while her height was spared. Deletion of Xq21.32q28 is a key factor leading to Turner syndrome-like phenotype such as rudimentary uterus and ovarian dysplasia.