Collagen (Coll), as the basic material of matrix scaffolds for cell growth, has been widely used in the field of tissue engineering and regenerative medicine. In this study, collagen protein was modified by L-lysine (Lys), and cross-linked by genipin (GN) to prepare the L-lysine-modified collagen (Lys-Coll-GN) scaffolds. Microstructure, pore size, porosity, stability and biocompatibility of Lys-Coll-GN scaffolds were observed. The results showed that the bond between L-lysine and collagen protein molecule was formed by generating amide linkage, and mouse embryo fibroblasts proliferation was not inhibited in the Lys-Coll-GN scaffolds. In the multiple comparisons of Coll-scaf- folds, Coll-GN scaffolds and Lys-Coll-GN scaffolds, Coll-scaffolds was the worst in mechanical characteristics while the highest in biodegradation rate. Compared to Coll-GN scaffolds, Lys-Coll-GN scaffolds had more fiber structure, higher interval porosity (P<0. 01). Although the tensile stress of Lys-Coll-GN scaffolds reduced significantly, its e- longation length extended when the scaffolds was fractured (P<0. 01). The percentage of Lys-Coll-GN scaffolds residual weight was lower than that of Coll-GN scaffolds after all the scaffolds were treated by collagenase for 5 days (P<0. 01). This study suggested that Lys-Coll-GN scaffold had good biocompatibility, and it improved the mechanical property and degradation velocity for collagen-based scaffold. This study gave a new predominant type of tissue engineering scaffold for the regenerative medicine.
Animals ; Biocompatible Materials ; chemistry ; Cell Proliferation ; Collagen ; chemistry ; Cross-Linking Reagents ; Fibroblasts ; cytology ; Iridoids ; chemistry ; Lysine ; chemistry ; Mice ; Porosity ; Tissue Engineering ; Tissue Scaffolds

Objective To discuss the clinical effects of transverse screw fixation of double columns in crescent pelvic fractures.Methods Twenty patients with crescent pelvic fractures hospitalized from December 2012 to December 2015 were reviewed retrospectively.There were thirteen male and seven female patients with the age ranging from 22 to 72 years (mean,39.5 years).Causes of injury were traffic accidents in fifteen patients,falling from high places in three and hitting by heavy objects in two.Time interval between injury and operation was 3-14 d.According to the Orthopedic Trauma Association (OTA) classification,all were classified as 61-B2 type.Reconstruct plate was used to stabilize the supperior pubic ramus fractures,and transverse double-column fixation with anterograde or retrograde screws was used for the posterior iliac crescent fractures.Duration of inserting screws,operation time,intraoperative blood loss and frequency of C-arm X-ray were recorded.Radiological and clinical outcomes were evaluated after operation.Results Duration of inserting screws ranged from 3 to 16 min (mean,5 min).Operation time ranged from 50 to 130 min (mean,80 min) and the intraoperative blood loss ranged from 200 to 550 ml (mean,280 ml).Frequency of C-arm X-ray in the surgery ranged from 1 to 5 times (mean,2 times).All screws were in the expected location,without any piercing out of the iliac bone dependent on the confirmation of X-ray and CT-scan after operation.According to the Matta and Tornetta radiological evaluation,the reduction was rated excellent in sixteen patients and good in four,with the excellent-good rate of 100％.There was no death,wound infection,sciatic nerve or superior gluteal nerve injury or deep venous thrombosis of lower extremities.At the followup,no nonunion,loss of reduction,and breakage of internal fixation occurred.Nineteen patients were followed up,and mean follow-up time was 19.6 months (range,3 to 36 months).Union was obtained in all patients in a period of 8-17 months (mean,11.2 months).According to the Majeed functional evaluation at the final follow-up,the outcome was rated excellent in seventeen patients and good in two,with the excellent-good rate of 100％.Conclusion Transverse screw fixation of double columns in crescent pelvic fracture of 61-B2 type can reduce the operation injury,decrease complications,and have good clinical results.

Objectives To identify the COL7A1 gene mutation in a recessive dystrophic epidermolysis bullosa (RDEB) family, and to perform prenatal diagnosis in the patient's offspring. Methods The genomic DNA, obtained from the patient and his wife, was used to screen all 118 exons of the type VII collagen gene (COL7A1) via polymerase chain reaction (PCR) followed by direct DNA sequencing of the PCR products. Fetal DNA was extracted from amniotic fluid of the patient's wife at the 15th week of gestation. PCR, direct DNA sequencing and restriction fragment length polymorphisms (RFLPs) were performed for prenatal diagnosis. Results The patient in this study was a compound heterozygote for a S48P missense mutation in exon 2 and an 11 base pair deletion (3625del11) leading to a premature termination codon (PTC) in exon 27, which are a novel combination of COL7A1 mutations in RDEB. The COL7A1 genotype of his wife was normal. In the fetus, the same deletion of 11 base pair (3625del11) was found in exon 27, but no mutation was found in exon 2. Thus, the fetus was predicted to be a clinically normal child with a carrier genotype. Seven months later, a clinically unaffected male infant was born and the prediction was confirmed. Conclusion We successfully performed the first DNA-based prenatal diagnosis in China in a family with Hallopeau-Siemens RDEB.

Objective To construct a colon cancer chemotherapy-resistant cell line COLO,and study its characteristics and its relationship with tumor stem cells.Methods We constructed two 5-fluorouraci (5-FU)-resistant colon cancer cell line COLO/5-FU-1 and COLO/5-FU-2, which were resistant to 0.1 0 μmol/ml and 0.20 μmol/ml 5-FU respectively through gradiently increased drug concentration.The cha-racteristics of 5-FU-resistant cell lines were compared with parental colon cancer cell line COLO related to proli-feration,colony forming ability,migration and invasion,sphere forming ability,expression of stemness genes and cross drug-resistance.Results In the cell viability assay,4 days after regular training,the absorbancy of colon cancer 5-FU-resistant cell lines COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were 0.61 ±0.1 3,0.54 ±0.07 and 0.41 ±0.09 respectively,with significant difference (F =63.43,P =0.033).With the increased concentration of 5-FU,5-FU-resistant cell lines presented increasing clonality. The cloning efficiency of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were (87.6 ±1 2.7)%,(65.3 ±9.7)% and (38.5 ±7.6)% respectively,with significant difference (F =33.64, P =0.01 7).In each high power field of vision,the cell numbers of migration through the basement membrane of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were 482 ±39,434 ±45 and 373 ±38 respectively;and the cell numbers of invasion through the basement membrane were 1 74 ±42,1 1 2 ± 31 and 87 ±29 respectively,with significant differences (F =1 09.61 ,P =0.009;F =67.31 ,P =0.032). Compared with parental colon cancer cell line COLO,5-FU-resistant cell lines had higher expression of stem-ness genes (F =47.31 ,P =0.042).5-FU-resistant cell lines were cross-resistant to other chemotherapeutic drugs such as mitoxantrone.For example,after incubation for 96 hours,inhibition rate of mitoxantrone to parent colon cancer cell line COLO was higher significantly than COLO/5-FU-1 and COLO/5-FU-2 (0.749 ± 0.042,0.423 ±0.024,0.342 ±0.01 8),with significant difference (F =1 2.61 ,P =0.028).The micro-sphere forming rates of COLO/5-FU-2,COLO/5-FU-1 and parental colon cancer cell line COLO were (8.90 ± 0.97)%,(6.20 ±0.75)% and (3.90 ±0.32)% respectively,with significant difference (F =1 64.32,P =0.006).Conclusion Colon cancer drug-resistant cell line COLO possess tumor stem cell-like characteristics, which are enriched in cancer stem cells.

OBJECTIVE: To apply combined non-invasive prenatal testing (NIPT), chromosomal karyotyping and chromosomal microarray for the screening and prenatal diagnosis of a fetus with supernumerary small marker chromosome (sSMC). METHODS: Standard NIFTY and full gene NIFTY kits were applied to detect free DNA (cfDNA) isolated from peripheral blood sample of a pregnancy woman. Amniocentesis was carried out for the woman for an abnormal NIPT result. G-banded karyotyping and single nucleotide polymorphism array (SNP array) were used to determine the karyotype and copy number variants in the fetus. The result was validated with a fluorescence in situ hybridization (FISH) assay. RESULTS: Both the standard NIFTY and full gene NIFTY indicated abnormal dup(chr12:707 334-33 308 759), for which the T score value of copy number anomaly in full gene NIFTY is 6.823, which is higher than the standard NIFTY's T-score value of 3.9535. The two NIFTY results were both above the normal threshold ± 3. Conventional G-banding analysis of amniocytes showed that the fetus has a karyotype of 47,XY,+mar. SNP-array delineated duplication of 12p (arr [hg19]12p13.33p11.1 (173 786_34 385 641)× 4, which was verified by FISH. Based on the above results, the fetus was diagnosed as a novel case of Pallister-Killian syndrome. CONCLUSION NIPT has a certain value for the prenatal detection of PKS. Combined use of multiple techniques can facilitate delineation of the source of sSMC.
Chromosome Disorders/genetics* ; Chromosomes, Human, Pair 12/genetics* ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy

OBJECTIVE: To explore the genetic basis for a child featuring developmental delay, intelligent disability and language deficit. METHODS: Peripheral blood samples of the child and her parents were collected for routine G-banding karyotyping analysis and single nucleotide polymorphism array (SNP array) detection. Amniotic fluid was also sampled from the mother for karyotyping analysis and SNP array detection. RESULTS: No karyotypic abnormality was found with the child and her parents. SNP array showed that the child has carried a 761.4 kb microdeletion at 16p11.2, while her mother has carried a 444.4 kb microduplication at 15q13.3. Her father's result was negative. Further analysis showed that the 15q13.3 microduplication was inherited from her maternal grandfather who was phenotypically normal. Prenatal diagnosis showed that the fetus has inherited the15q13.3 microduplication from its mother. CONCLUSION The child has carried a de novo 16p11.2 microdeletion, which overlaps with 16p11.2 microdeletion syndrome region, in addition with similar clinical phenotypes. The 16p11.2 microdeletion probably underlies her abnormal phenotype.
Child ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 16 ; Developmental Disabilities/genetics* ; Female ; Fetus ; Humans ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis

Objective: To explore the genetic basis of a child with developmental delay and intellectual disability. Methods: Peripheral blood samples of the child and his parents were collected for routine G-band karyotyping analysis and single nucleotide polymorphism array (SNP array) assay. Amniotic fluid sample was collected during the next pregnancy for prenatal diagnosis. Results: No karyotypic abnormality was found in the child and his parents. SNP array showed that the child has carried a 855.3 kb microduplication in 15q11.2. His mother carried the same duplication but had no phenotypic anomaly. No microdeletion/microduplication was found in his father. Upon prenatal diagnosis, no abnormalities was found with the chromosomal karyotype and SNP array result of the fetus. Conclusion 15q11.2 microduplication may result in developmental delay and intellectual disability, for which CYFIP1 may be a candidate gene. However, the duplication may increase the risk but with a low penetrance. This should attract attention during clinical consultation.

OBJECTIVE: To explore the pathogenetic mechanism of cystic meningioma and the key to its diagnosis and operation. METHODS: Clinical data of 8 patients with cystic meningioma were analyzed retrospectively. The occuring position and cure rate of cystic meningioma were compared with those of solid meningioma in the corresponding period. RESULTS: All patients were subjected to total resection of tumor parenchyma and cystic wall. No operative death and severe complications occurred. The incidence of uppertentorial region of cystic meningioma was higher than that of solid meningioma, and the difference was significant (χ(2)=2.618, P<0.05). The cure rate was not significantly different between cystic meningioma and solid meningioma (χ(2)=0.010, P>0.05). CONCLUSION Removing tumor totally is the key to preventing its recurrence.
Adult ; Aged ; Arachnoid Cysts ; complications ; diagnosis ; surgery ; Female ; Humans ; Male ; Meningeal Neoplasms ; diagnosis ; surgery ; Meningioma ; classification ; complications ; diagnosis ; surgery ; Middle Aged ; Retrospective Studies

Objective: To explore the association between processed food consumption and anxiety symptoms among college students in Yunnan Province, so as to provide a reference for the prevention and treatment of anxiety symptoms in this population. Methods: A cluster random sample of 2 515 first year students from two universities in Yunnan Province was selected to carry out a longitudinal investigation which included a baseline survey (November 2021, T1) and three follow up visits (June 2022, T2; November 2022, T3; June 2023, T4). The food frequency questionnaire was administered to assess processed food consumption, and the Depression Anxiety Stress Scale-21 (DASS-21, Chinese version) was used to evaluate anxiety symptoms. A generalized estimation equation model was used to analyze the relationship between processed food consumption and anxiety symptoms. Results: The detection rates of T1-T4 anxiety symptoms among college students in Yunnan Province were 29.70%, 36.70%, 37.69% and 38.73 %, respectively, and the corresponding anxiety symptom scores were 4(0,8), 4(0,10), 4(0,12), 2(0,14). After controlling for demographic variables and confounding factors in the generalized estimation equation model, a statistically significant association was found between consumption of carbonated beverages ( β=0.06, 95%CI =0.03-0.08), and other processed snacks ( β= 0.04 , 95%CI =0.01-0.07) ( P <0.05). The stratified analysis by gender showed that the consumption of carbonated beverages ( β=0.08, 95%CI =0.05-0.12) and fast food ( β=0.03, 95%CI =0.00-0.06) was significantly associated with anxiety symptoms in female college students ( P <0.05). There was no significant association between processed food consumption and anxiety symptoms in male college students ( P >0.05). Conclusions Processed food consumption by college students in Yunnan Province may increase the risk of anxiety symptoms, particularly among female students. There is a need to strengthen guidance in respect to processed food consumption, so as to prevent and treat anxiety symptoms.

Objective To develop a diagnostic model based on deep neural network for intelligent discrimination of thyroid function. Methods A total of 1616 patients ( 283 males, 1333 females, average age:52 years) who underwent thyroid imaging between May 2016 and June 2018 were selected. According to the clinical diagnosis, the 1616 cases included 299 normal thyroid cases, 876 hyperthyroidism cases and 441 hypothyroidism cases. Feature extraction and learning training were performed on 1000 training set sam-ples by two deep neural network models ( AlexNet;deep convolution generative adversarial networks ( DCGAN) ) using deep learning algorithm. Performance verifications were implemented on 616 test set samples. The con-sistency between the verification results of the two models and the clinical diagnosis was analyzed by Kappa test. Meanwhile, the time advantage of the intelligent diagnosis models was analyzed. Results The average diagnostic time of AlexNet model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 82.29％(79/96), 94.62％(369/390), 100％(130/130), respectively. The Kappa value between results of AlexNet model and clinical diagnosis was 0.886 ( P<0.05) . The average di-agnostic time of DCGAN model was 1 s/case, and the classification accuracy for normal thyroid, hyperthy-roidism, hypothyroidism were 85.42％(82/96), 95.64％(373/390), 99.23％(129/130), respectively. The Kappa value between results of DCGAN model and clinical diagnosis was 0.904 ( P<0.05) . Conclusion The deep neural network intelligent diagnosis model can quickly determine the functional status of thyroid gland in thyroid imaging, and it has a high recognition accuracy, thus providing a new method for thyroid image review.