Collagen, a vital matrix protein for various tissue and functions in animals, is widely applied in biomaterials. In type Ⅰ collagen, missense mutations of glycine (Gly) in the Gly-Xaa-Yaa triplet of the triple helix are a major cause of osteogenesis imperfecta (OI). Clinical manifestations exhibit marked heterogeneity, spanning a broad disease spectrum from mild skeletal fragility (Type Ⅰ) to severe limb deformities (Type Ⅲ) and perinatal lethal forms (Type Ⅱ). This study utilized recombinant collagen as a model to further elucidate whether Gly→Ala/Val mutations at the N-terminus of the integrin-binding sequence GFPGER affect collagen structure and function, and to explore the underlying mechanisms by which missense mutations impact the biological function of collagen. By introducing Ala and Val substitutions at seven Gly positions N-terminal to the GFPGER sequence, we systematically assessed the effects of these amino acid replacements on the triple-helical structure, thermal stability, integrin-binding ability, and cell adhesion of recombinant collagen. All constructs formed a stable triple-helix structure, with slightly compromised thermal stability. Gly→Val substitutions increased the susceptibility of recombinant collagen to trypsin, which suggested local conformational perturbations in the triple helix. In addition, Gly→Val substitutions significantly reduced the integrin-binding affinity and decreased HT1080 cell adhesion, with the effects stronger than Gly→Ala substitutions. Compared with Gly→Ala substitutions, substitution of Gly with the larger residue Val had enhanced negative effects on the structure and function of recombinant collagen. These findings provide new insights into the molecular mechanisms of osteogenesis imperfecta and offer theoretical references and experimental foundations for the design of collagen sequences and the development of collagen-based biomaterials.
Recombinant Proteins/biosynthesis* ; Glycine/genetics* ; Humans ; Osteogenesis Imperfecta/genetics* ; Integrins/metabolism* ; Collagen/metabolism* ; Collagen Type I/metabolism* ; Amino Acid Substitution ; Mutation ; Mutation, Missense

ObjectiveTo observe the effects of Agrimoniae Herba and Coptidis Rhizoma（XHC-HL） on the proliferation， migration， invasion， apoptosis， and autophagy of colorectal tumor cells and explore the mechanism of Agrimoniae Herba and Coptidis Rhizoma in inhibiting colorectal cancer by regulating chaperone-mediated autophagy（CMA）. MethodXHC-HL-containing serum was prepared， and human colorectal cancer cell lines HT29 and LOVO were cultivated and divided into blank group （20% blank serum）， low XHC-HL groups （5%， 10%， 20% drug-containing serum）， and a positive control group using 5-fluorouracil （5-FU）. Cell viability was measured using the cell counting kit-8 （CCK-8） method. Cell proliferation ability was assessed through 5-ethynyl-2′-deoxyuridine （EdU） staining. Scratch assays and Transwell migration tests were conducted to evaluate cell migration ability， and Transwell invasion assays were used to assess cell invasion ability. Apoptosis rates were determined by flow cytometry， and the impact of XHC-HL on yeast Atg6 homologous 1 （Beclin-1）， microtubule-associated protein1 light chain3 Ⅱ （LC3 Ⅱ）， and p62 was analyzed via Western blot. The influence of XHC-HL on CMA-related proteins and mRNA was examined using Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared to that in the blank group， the vitality of colorectal cancer cells HT-29 and LOVO was significantly inhibited in XHC-HL groups and the 5-FU group （P<0.01）. Compared to that in the blank group， the proliferation of colorectal cancer cells HT-29 and LOVO was significantly inhibited in XHC-HL groups and the 5-FU group （P<0.05， P<0.01）. Compared to the blank group， there was a significant reduction in cell migration rates （P<0.01）， the number of cells migrating to the lower chamber （P<0.01）， and the number of invasive cells （P<0.01）， as well as and an increase in cell apoptosis （P<0.01） in XHC-HL groups and the 5-FU group （P<0.01）. Western blot results indicated that compared to that in the blank group， the expression levels of Beclin-1 and LC3 Ⅱ were increased（P<0.05，P<0.01）in XHC-HL groups and the 5-FU group， while the p62 levels were decreased（P<0.05，P<0.01）. Furthermore， the protein expression levels of lysosomal associated protein 2A （LAMP2A）， heat shock cognate protein 70 （HSC70）， and heat-shock protein 90 （HSP90） in XHC-HL groups and the 5-FU group were decreased to varying extents， whereas the expression levels of glyceraldehyde-3-phosphate dehydrogenase （GAPDH） were elevated （P<0.01）. The Real-time PCR results showed that compared to those in the blank group， the mRNA levels of LAMP2A， HSC70， and HSP90 were downregulated in XHC-HL groups and the 5-FU group， and the mRNA expression level of GAPDH was upregulated （P<0.01）. ConclusionXHC-HL can inhibit the proliferation， migration， and invasion of colorectal cancer cells by suppressing CMA activity， thus inducing autophagy and promoting apoptosis.

【Objective】 To explore the correlation between the severity of jaundice in ABO hemolytic disease of fetus and newborn (HDFN) and multiple indicators, in order to achieve accurate diagnosis, timely treatment, and prevent complications in ABO HDFN. 【Methods】 From March 2020 to February 2023, a total of 283 ABO HDFN in our hospital were classified into mild, moderate and severe groups based on total bilirubin levels. The differences in basic data, relevant laboratory indicators and the agglutination intensity of the three hemolysis tests were analyzed and compared. 【Results】 In the severe group, 75% (12/16) were male infants, which was significantly higher compared to 37.5% (57/152) in the moderate group (P<0.05). The values (mg/dL) of transcutaneous jaundice in the mild, moderate and severe groups were:forehead 9.88±1.93 vs 12.34±2.01 vs 15.56±2.69, face 10.25±2.27 vs 13.28±2.32 vs 15.99±2.86, chest 9.67±2.16 vs 12.51±2.11 vs 15.33±2.36, respectively. The values(µmol/L) of direct bilirubin (DBIL) and indirect bilirubin (IBIL) were 9.87±2.06 vs 11.90±2.59 vs 16.12±4.73, and 159.36±37.55 vs 252.98±30.52 vs 353.76±55.68, respectively, and the differences between the mild, moderate and severe groups were statistically significant (all P<0.05) . The creatine kinase (CK) values(unit/L) of mild, moderate and severe groups were 664.15±498.26 vs 500.51 ±451.63 vs 402.63±224.27, and the difference between the mild group and the other two groups was statistically significant (P<0.05) . There was no statistically significant difference in the agglutination intensity of the three hemolysis tests among HDFN with different severity of jaundice. 【Conclusion】 Clinicians can predict the severity of jaundice in light of gender, transcutaneous jaundice, DBIL, IBIL and CK of ABO HDFN, for further and graded treatment, so as to avoid the sequelae or even life-threatening consequences caused by ABO HDFN.

BACKGROUND: The effect of arteriosclerotic intracranial arterial vessel wall enhancement (IAVWE) on downstream collateral flow found in vessel wall imaging (VWI) is not clear. Regardless of the mechanism underlying IAVWE on VWI, damage to the patient's nervous system caused by IAVWE is likely achieved by affecting downstream cerebral blood flow. The present study aimed to investigate the effect of arteriosclerotic IAVWE on downstream collateral flow. METHODS: The present study recruited 63 consecutive patients at the Second Hospital of Hebei Medical University from January 2021 to November 2021 with underlying atherosclerotic diseases and unilateral middle cerebral artery (MCA) M1-segment stenosis who underwent an magnetic resonance scan within 3 days of symptom onset. The patients were divided into 4 groups according to IAVWE and the stenosis ratio (Group 1, n = 17; Group 2, n = 19; Group 3, n = 13; Group 4, n = 14), and downstream collateral flow was analyzed using three-dimensional pseudocontinuous arterial spin labeling (3D-pCASL) and RAPID software. The National Institutes of Health Stroke Scale (NIHSS) scores of the patients were also recorded. Two-factor multivariate analysis of variance using Pillai's trace was used as the main statistical method. RESULTS: No statistically significant difference was found in baseline demographic characteristics among the groups. IAVWE, but not the stenosis ratio, had a statistically significant significance on the late-arriving retrograde flow proportion (LARFP), hypoperfusion intensity ratio (HIR), and NIHSS scores ( F = 20.941, P <0.001, Pillai's trace statistic = 0.567). The between-subject effects test showed that IAVWE had a significant effect on the three dependent variables: LARFP ( R2 = 0.088, F = 10.899, P = 0.002), HIR ( R2 = 0.234, F = 29.354, P <0.001), and NIHSS ( R2 = 114.339, F = 33.338, P <0.001). CONCLUSIONS: Arteriosclerotic IAVWE significantly reduced downstream collateral flow and affected relevant neurological deficits. It was an independent factor affecting downstream collateral flow and NIHSS scores, which should be a focus of future studies. TRIAL REGISTRATION ChiCTR.org.cn, ChiCTR2100053661.
Humans ; Constriction, Pathologic/pathology* ; Magnetic Resonance Imaging/methods* ; Middle Cerebral Artery/pathology* ; Tomography, X-Ray Computed

This study was designed to investigate the effect of salidroside on Sjogren's syndrome in mice and its mechanism.Mice in negative control group and model group were given normal saline intragastric administration,mice in 3 salidroside groups were given salidroside intragastric administration(doses of 20,40 and 80 mg/kg),and mice in positive control group were given hydroxychloroquine sulfate(100 mg/kg)intragastric administration once a day.After continuous intragastric administration for 8 weeks,water intake and saliva flow rate were detected,infiltrated submandibular gland lymphocytes were evaluated,the levels of IL-17,IL-10,NF-κB P65 and IκBα and the ratio of Th17/Treg cells were detected.In the model group,the acinus atrophied with unclear margin and decreasing in number,and the lymphocyte infiltration were observed and lymphocyte focus was formed.After the intervention with salidroside and hydroxychloroquine sulfate,the degree of acinus lesions was relieved to a certain extent,and the lymphocyte infiltration was reduced.Compared with negative control group,water intake,salivary flow rate,submandibular gland index,IL-10 and IκBα levels were decreased in other groups,while lymphocyte infiltration,the levels of IL-17,IL-17/IL-10,NF-κB P65 and NF-κB P65/IκBα were increased(P<0.05).Compared with model group,water intake,salivary flow rate,submandibular gland index,IL-10 and IκBα levels were increased in each salidroside dose group,while submandibular gland lymphocyte infiltration,the levels of IL-17,IL-17/IL-10,NF-κB P65 and NF-κB P65/IκBα decreased in a dose-dependent manner(P<0.05).Compared with the negative control group,the proportion of Th17 cells in the serum of model group was increased,and the proportion of Treg cells was decreased,while salidroside in all doses could reverse these changes(P<0.05).Taken together,salidroside alleviates submandibular gland inflammatory responses by mediating Th17/Treg immune balance and inhibiting NF-κB P65/IκBα,thus playing a therapeutic role in SS treatment.

Objective:To explore the surgical technique and therapeutic effect of flap wrapping repair of soft tissue defect after digit degloving injury with free arm medial perforator flap.Methods:From October 2009 to December 2020, 15 patients with soft tissue defect after digit degloving injury were repaired with free arm medial perforator flaps. The patients were 9 males and 6 females aged 22 to 50 years old, with an average of 32 years old. Injury mechanism: 13 cases of machine strangulation and 2 cases of machine crushing. Plane of sleeve avulsion injury: at the distal metacarpophalangeal joint of thumb in 2 cases, at the middle and distal segment of single finger of 2nd to 5th fingers in 4 cases, and at distal metacarpophalangeal joint of 2nd to 5th fingers in 9 cases. Seven cases were in left hand and 8 in right hand. Emergency surgery was performed in 2 cases and scheduled surgery in 13 cases. The flap was designed in a shape of a long strip, and the size of the flap was 5.0 cm×2.5 cm-14.0 cm×3.5 cm. During the operation, the artery of the flap pedicle was anastomosed with the palmar digital proper artery or common digital artery of the recipient digit, the subcutaneous vein or the companion vein of the artery was anastomosed with the dorsal or palmar digital subcutaneous vein, and the medial brachial cutaneous nerve carried in the flap was anastomosed with the stump of palmar digital proper nerve. All the donor sites were directly sutured. After the surgery, follow-up visits were conducted regularly at outpatient clinic, or via telephone and WeChat review or by home visit. The flap appearance, sensation and function recovery of digital joints were observed together with the patient satisfaction. Results:All flaps survived successfully after surgery. Six patients with degloving defect of the whole digit had encountered poor wound healing caused by distal phalangeal necrosis. The second stage stump trimming was performed to keep the digit to the distal end of the middle segment. Two cases of thumb and 2 cases of middle finger suffered further burning and worn tears at 7-9 months after surgery and self-healed. The follow-up period was 6-28 (average 16) months. The flap was soft without bloating. The sensation recovered to S 2-S 3. The shape of fingers was good, and no secondary flap surgery was necessary. The overall movement of interphalangeal joints was poor. According to the evaluation standard of Michigan hand function questionnaire, 15 patients achieved very satisfactory with the overall appearance and function of hands. The linear scars at donor site were hidden without complications such as tenderness and contraction. Conclusion:The free medial perforator flap of the arm is easy to design and thin, hence does not affect the shape of a digit. So, it is ideal for the repair of soft tissue defect after digit degloving injury.

Objective:to investigate the therapeutic effect of low molecular weight heparin combined with average volumeassuredpressuresupport (AVAPS) on patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) complicated with type Ⅱ respiratory failure.Methods:A total of 82 patients with AECOPD combined with type Ⅱ respiratory failure in the Second People′s Hospital of Xindu District of Chengdu from February 2018 to April 2020 were selected as the research objects, and they were randomly divided into two groups with 41 cases in each group. The control group was given AVAPS mode, and the observation group was given low molecular weight heparin combined with AVAPS mode. The arterial partial pressure of oxygen (PaO 2), arterial carbon dioxide (PaCO 2), and forced vital capacity (FVC), forced expiratory volume in the first second (FEV 1), maximum peak expiratory flow rate (PEF), interleukin (IL)-6, IL-8, tumor necrosis factor-(TNF-α), 16 kDa Clara cell protein (CC16), surfactant protein D(SP-D), adiponectin (APN), superoxide dismutase (SOD), D-dimer, fibrinogen before and after treatment were compared between the two groups and the incidence of adverse reactions were observed. Results:After treatment, the levels of FEV 1, FVC, PEF, PaO 2 in the observation group were higher than those in the control group: (1.78 ± 0.29) L vs. (1.47 ± 0.25) L, (2.47 ± 0.29) L vs.(2.20 ± 0.25) L, (5.14 ± 0.37) ml/s vs. (4.69 ± 0.35) ml/s, (88.37 ± 10.52) mmHg (1 mmHg = 0.133 kPa) vs. (80.16 ± 9.87) mmHg; and the level of PaCO 2 was lower than that in the control group: (65.07 ± 6.71) mmHg vs. (70.84 ± 6.50) mmHg; and the differences were statistically significant ( P<0.05). After treatment, the levels of IL-6, IL-8 and TNF-α in the observation group were lower than those in the control group: (0.47 ± 0.09) ng/L vs. (0.58 ± 0.10) ng/L, (64.37 ± 7.25) ng/L vs. (88.24 ± 8.34) ng/L, (45.37 ± 4.63) ng/L vs. (66.31 ± 4.92) ng/L; and the levels of SOD and APN were higher than those in the control group: (92.37 ± 10.85) U/mg vs. (76.13 ± 9.84) U/mg, (13.94 ± 0.76) mg/L vs. (11.58 ± 1.21) mg/L; and the differences were statistically significant ( P<0.05). After treatment, the level of CC16 in the observation group was higher than that in the control group: (114.78 ± 12.15) μg/L vs. (107.41 ± 11.06) μg/L; while the levels of SP-D, D-dimer and FIB were lower than those in the control group: (93.24 ± 9.85) μg/L vs. (103.25 ± 10.78) μg/L, (0.58 ± 0.07) mg/L vs. (0.79 ± 0.11) mg/L, (1.98 ± 0.29) g/L vs. (2.56 ± 0.34) g/L; and the differences were statistically significant ( P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:Low-molecular-weight heparin combined with AVAPS mode in the treatment of AECOPD complicated with type Ⅱ respiratory failure can significantly improve the lung inflammation and coagulation function in patients, adjust blood gas analysis and CC16, SP-D levels, and promote the recovery of patients′ lung function.

【Objective】 To compare the platelet apheresis donation rate via telephone and WeChat appointment in Shijiazhuang area during the COVID-19 epidemic, so as to give suggestions for optimizing blood donation appointment in major public events. 【Methods】 The number and appointment type of apheresis donors from January to March 2019(the control) vs January to March 2020(study group 1) vs January to March 2021(study group2) in Shijiazhuang area were collected. A total of 1 204 and 1 305 questionnaires were distributed to two study groups to analyze their acceptance of telephone and WeChat appointment. 【Results】 More turnout donors preferred WeChat appointment, relative to telephone calls, during January to March 2020 and 2021. But some inactive donors preferred appointment given by telephone calls. 【Conclusion】 Different appointment methods should be adopted to recruit blood donors according to the targets of blood centers, in the event of national or local COVID-19 epidemic.

OBJECTIVE: To assess the clinical value of non-invasive prenatal testing (NIPT) for the screening of trisomy and copy number variations (CNVs) of chromosomes 21, 18 and 13. METHODS: From January 2015 to December 2019, 40 628 pregnant women underwent NIPT testing using high-throughput sequencing and bioinformatics analysis to test the cell-free fetal DNA in maternal plasma. High-risk pregnant women underwent invasive prenatal diagnosis, while low-risk ones were followed up by telephone. RESULTS: The three most common indications included intermediate risk of serological screening, high risk of serological screening and advanced maternal age. Among all pregnant women, 257 cases were detected as trisomy 21, 18 and 13 (170, 49 and 38 cases, respectively). 227 cases chose invasive prenatal diagnosis, with respectively 122, 28 and 10 cases confirmed. The positive predictive value (PPV) was 81.33% (122/150), 65.12% (28/43), 29.41% (10/34), respectively. Two false negative cases of trisomy 18 were found during follow-up. Meanwhile, NIPT has detected 46 cases (15, 16 and 15 cases, respectively) CNVs on chromosomes 21, 18 and 13, among which 37 cases underwent invasive prenatal diagnosis. There were 5, 3 and 5 positive cases, which yielded a PPV of 41.67% (5/12), 25%(3/12) and 33.33%(5/15), respectively. Two other chromosome CNVs were accidentally discovered among the false positive samples. CONCLUSION The incidence of chromosomal abnormalities in the serological screening high-risk group was 52.02%, which was significantly higher than other groups. NIPT has a high sensitivity and specificity for the screening of trisomies 21, 18 and 13, while its accuracy for detecting CNVs of chromosomes 21, 18 and 13 needs to be improved. As a screening method, NIPT has a great clinical value, though there are still limitations of false positive and false negative results.Comprehensive pre- and post-test genetic counseling should be provided to the patients.
Aneuploidy ; Chromosome Disorders/genetics* ; Chromosomes ; DNA Copy Number Variations ; Down Syndrome/genetics* ; Female ; Humans ; Pregnancy ; Prenatal Diagnosis ; Trisomy/genetics* ; Trisomy 18 Syndrome/genetics*

Objective:To examine the effect of childhood abdominal obesity on abdominal obesity in adulthood.Methods:Based on data from the "China Health and Nutrition Survey (CHNS)" 1993-2011, the participants who were followed up at least once in childhood (7-17 years old) and adulthood (≥18 years old) were the potential eligible ones for the inclusion. The first follow- up data in childhood and the last follow-up data in adulthood were finally included in this study, and there were 1 366 participants (males: 61.4 %). Partial correlation analysis was used to examine the correlation of waist circumference in childhood with adult waist circumference. Cox regression model was used to examine the effect of abdominal obesity in childhood on abdominal obesity in adulthood. Results:The median follow-up of this cohort was 10.4 years. 36.4 % (28/77) of children with abdominal obesity remained abdominally obese in adulthood. The coefficient of partial correlation was moderate for association of childhood waist circumference with adult waist circumference ( r=0.32, P<0.001). The risk of abdominal obesity in adulthood was significantly increased in abdominal obese children ( HR=7.54, 95 %CI: 4.91-11.58) compared with children with normal waist circumference. Conclusion:Children with abdominal obesity significantly increases the risk of abdominal obesity in adulthood.