The role of Sonic Hedgehog (SHH)signaling pathway in embryonic lung development has been recognized.What's more,it also plays a key role in postnatal development and maintenance of tissue or organ integrity and function.In recent years, studies have found that SHH signaling pathways are involved in the lung repairing process, suggesting that SHH signaling pathways may play a role in pulmonary fibrosis.This paper reviews the effect of the SHH signaling pathway in pulmonary fibrosis.

Different maternal positions and movement are utilized gradually in clinic while today it appears that the majority of women in the world birth in supine position or lithotomy position. To know the current situation and progress of the studies concerning different maternal positions and movement during labour and to explore the current relevant problems in clinic. The search strategy was designed to identify the relevant literature. The search was limited to the last 20 years as current literature was sought. Thirty eight papers were identified as relevant and included in this literature review. The majority studies is concerning about the outcome of mother and fetus. There is no unitized clinical pathway currently, and the objects implemented are limited. There is no study related to the requirement of implementing different maternal positions and movement during labor in clinic and the effect of pelvic floor function after parturition. The positive effect of implementing different maternal positions and movement during labor can be sure for the process of parturition and the outcome of mother and baby. Childbirth in different maternal positions and movement is the trend in the future. Therefore, how to utilize different maternal positions and movement during labor well, it is still a huge challenge for medical staff. And more convincing studies and information are needed as evidence in clinic.

BACKGROUND: There are different methods to isolate and culture human nucleus pulposus cells, and the differences in digestive enzymes components and digestion time quite are significant. So how to rapidly and efficiently harvest human nucleus pulposus cells has become a research hotspot. OBJECTIVE: To optimize the digestive enzymes components and digestion methods for the preparation of human nucleus pulposus cells. METHODS: Nucleus pulposus tissue specimens were selected from three adult discs in the Department of Orthopedics, China-Japan Union Hospital of Jilin University. The acute traumatic disc tissues that outstanding to the spinal canal were taken under aseptic conditions, and then the peripheral white annulus and jel y-like nucleus pulposus in the center could be seen. According to different mixed enzyme concentration ratio, the samples were divided into two groups. The enzyme Ⅰ group was treated with 0.2% Ⅱ col agenase; and the mixed enzymeⅡ group was digested with 0.25% trypsin for 30 minutes, and then treated with 0.2% Ⅱ col agenase. According to digestion time, each group was divided into three subgroups: 2 hours group, 4 hours group, and overnight group. Final y, suspended cel volume was decided as 2 mL to count cells. Dulbecco’s modified Eagle’s medium containing fetal bovine serum was used for cel culture in vitro. Trypan blue staining was performed to count total cel number and ratio of living cells. Methylthiazolyldiphenyl-tetrazolium bromide assay was used to detect the growth curve of nucleus pulposus cells. RESULTS AND CONCLUSION: Based on the two digestion enzyme concentration, the number of digested cells in the enzyme Ⅰ group was larger than that in the enzyme Ⅱ group after digested for 2 and 4 hours, but the difference was not significant (P > 0.05). Overnight, cellsurvival rate was decreased in the enzyme Ⅰ group after digested for 2 and 4 hours when compared with the enzyme Ⅱ group, and the difference was significant (P < 0.05). After digested for 4 hours, tissue blocks disappeared, and the number of cells reached maximum. The results indicate that enzyme Ⅰgroup composite with Ⅱ col agenase is benefit for the separation of nucleus pulposus cells, and the digestion time is appropriate to 4 hours. This condition has the advantages of simple operation, high efficiency and low cost, and it considered that digestion of nucleus pulposus tissues with 0.2% Ⅱ col agenase for 4 hours is the best condition to obtain nucleus pulposus cells.

BACKGROUND:Advancement in bioengineering based upon tissue engineering techniques may offer the possibility of repairing degenerative intervertebral disc.
OBJECTIVE:To summarize the research progress in the scaffolds of tissue engineered intervertebral disc.
METHODS:A computer-based retrieval was performed to search manuscripts describing tissue engineered intervertebral disc scaffolds published between January 1st, 1900 and December 31st, 2012 in PubMed database with the key words of“tissue engineering, intervertebral disc, scaffold”in English.
RESULTS AND CONCLUSION:Scaffold is an important part of tissue-engineered research. There are three kinds of materials for intervertebral disc scaffolds:natural biomaterials, synthetic materials, and composite materials. A variety of scaffold materials have their own advantages and disadvantages. Up to now, none of these scaffold materials is accepted as the most suitable one. The selection of scaffold materials is stil to be further studied. The study and development of nanoscale biomaterials is an inevitable trend. Otherwise, with the help of bionics, improving scaffolds is also an inexorable trend in progress of simulating human intervertebral disc. Furthermore, injectable scaffold is also an research hot spot, and the selection range of injectable scaffold materials mainly focuses on chitosan, typeⅡcolagen,hyaluronic acid,fibrin,elastin,and alginate.C urrently, studies on chitosan as a scaffold material are relatively more.

Objective To investigate the effect of epidermal growth factor (EGF) on the expression of Matrix metalloproteinase-9 (MMP-9) and the signalling pathways involved in the trophoblast cell line JEG-3. Methods The JEG-3 trophoblast cell line was used in this study. (1) JEG-3 cells were cultured with various concentrations of EGF (0, 1, 10,20 ng/ml) for 24 hours and the expression of MMP-9 was tested by western blotting and reverse transcription PCR (RT-PCR). (2) Western blotting and RT-PCR were also used to investigate the expression of MMP-9 expression after incubation for 0,4,12 and 24 hours with EGF treatment (10 ng/ml) in JEG-3 cells. (3) According to the different added ingredients, JEG-3 cells were divided into some groups: control group (without EGF), EGF group (exposure to l0ng/ml EGF),EGF + inhibitors group (exposure to 10 ng/ml EGF +20 ng/ml SB203580 or exposure to 10 ng/ml EGF + 10ng/ml U0126) inhibitors group (exposure to 20 ng/ml SB203580 or exposure to 10 ng/ml U0126). Western blotting were used to investigate the expression levels of MMP-9, nuclear factor kappa B (NF-kB) ,p38MAPK, phospho-p38MAPK (p-p38MAPK) , extracellular -signal regulated kinase (ERK) and phospho-ERK (p-ERK) protein in JEG-3 cells after incubation for 24 hours. Results (1) The profiles of MMP-9mRNA were increased by various concentrations of EGF (0, 1 , 10, 20 ng/ml) in JEG-3 cells after 24hculture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0. 567 ±0. 056) , 10ng/ml of EGF (1. 392 ±0. 133) , 20 ng/ml of EGF (1. 971 ±0. 067) were significantly higher respectively (P <0. 05) , compared with 0 ng/ml of EGF treatment (0. 166 ±0. 015). Similarly, MMP-9 mRNAs were also increased with the increasing incubation time. Compared to EGF (10 ng/ml) stimulation for 0 h (0.253 ±0.044), the MMP-9 mRNA profiles were 0. 470 ±0. 026, 1.061 ±0. 115, 1. 453 ±0. 180 for 4,12 and 24 hours, respectively (P < 0. 05). (2) In accordance to the mRNA profiles, the expression of MMP-9 protein was also increased by different concentrations of EGF (0,1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The abundance of MMP-9 protein in the three groups was 0. 043 ±0. 012, 0. 085 ±0. 008, 0. 142 ±0. 015, with a significantly higher expression, compared with 0 ng/ml of EGF treatment (0. 004 ±0.001, P < 0.05) respectively. Similarly, MMP-9 proteins were also increased with the increasing incubation time. Compared to EGF(10 ng/ml) stimulation for 0 h (0. 030 ±0. 009) , the profiles of MMP-9 protein were 0. 137 ± 0. 010, 0. 240 ± 0. 010, 1.240 ±0.061 for 4, 12 and 24 hours, respectively (P < 0.05). (3) Both p38MAPK and ERK signalling pathways were activated by EGF in JEG-3 cells. The expression of p-p38MAPK was significantly higher (without or with 10 ng/ml EGF, 234. 1 ± 4. 1 vs.260. 9 ± 2. 5 , P < 0. 05) , however, the p38 MAPK inhibitor SB203580 markedly suppressed the increase in p-p38MAPK content induced by EGF(227. 9 ±2. 4 vs. 260. 9 ±2. 5, P<0. 05). Similarly, the expression of p-ERK was significantly higher with EGF treatment (812. 2 ±3. 5) vs. without EGF group (453.4±5.8) (P <0. 05) , while the ERK inhibitor U0126 significantly inhibited the increased p-ERK content in response to EGF treatment (71. 0 ± 1. 2 vs. 812. 2 ± 3. 5, P < 0. 05) . (4) The p38MAPK inhibitor SB203580 significantly reduced the expression of EGF-induced MMP-9 (0. 645 ± 0. 270 vs. 1. 476 ± 0. 452, P < 0. 05)and NF-kB (0.530 ± 0.026 vs. 0.959 ± 0. 017, P < 0. 05) . (5) The ERK inhibitor U0126 also significantly reduced the expression of EGF-induced MMP-9 (0. 623 ±0. 030 vs. 2. 112 ±0. 056, P <0. 05)and NF-kB (0. 325 ± 0. 082 vs. 0. 939 ± 0. 153, P < 0. 05). Conclusion EGF induced the expression of MMP-9 in a time and dose-dependant manner in JEG-3 cells. EGF enhanced MMP-9 expression through the activation of p38MAPK and ERK signalling pathways in JEG-3 cells.

Objective To investigate the effect of transforming growth factor β1 (TGF-β1) on the expression of matrix metalloproteinase 9 (MMP-9),tissue inhibitor of metalloproteinase 1 (TIMP-1),nuclear factor kappa B(NF-κB) and the possible signalling pathways in human amniotic cells WISH.Methods The WISH cell line was cultured.WISH cells were added with TGF-β1 of different concentrations (0,2,10 and 20 ng/ml,respectively) for 24 hours.Then,reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot.Results (1) The profile of TIMP-1 mRNA (0.413 ±0.036,0.623 ±0.058,1.392 ±0.124,1.387 ±0.102) in WISH cells elevated when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).In accordance with TIMP-1 mRNA,the expression of TIMP-1 also elevated with the increase of TGF-β1 (0.357 ± 0.031,0.596 ± 0.048,1.243 ± 0.097 and 1.359 ± 0.121,respectively).And when 2,10 or 20 ng/ml of TGF-β1 was added,the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-β1 was added(P ＜ 0.05).(2)In contrast with TIMP-1,MMP-9 mRNA (1.325 ±0.056,0.987 ±0.081,0.610 ±0.034,0.347 ±0.023) in WISH cells decreased when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).The MMP-9 protein (1.119 ±0.064,1.008 ±0.052,0.578 ±0.041,0.401 ±0.015) also decreased with the increase of TGF-β1.And when 2,10 or 20 ng/ml of TGF-β1 was added,the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-β1 was added (P ＜ 0.05).(3) The NF-κB protein (1.423 ±0.065,1.116 ± 0.045,0.796 ± 0.041,0.359 ± 0.021) was significandy reduced with the increase of TGF-β1 (0,2,10,20 ng/ml; P ＜ 0.05).Conclusions The mRNA and protein expression of TIMP-1 decreased when TGF-β1 was low in WISH cells,whereas those of MMP-9 elevated when TGF-β1 was low.The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane.And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.

Objective To investigate the cognitive status of pregnant women on labor in liberal position and demand investigation, and analyze the reason and influencing factors of pregnant women choosing labor in liberal position. Methods Totally 177 pregnant women from the Third Affiliated Hospital of Guangzhou Medical University were chosen by convenience sampling method. They were investigate by self-designed questionnaire in order to get their information. Results The cognition of pregnant women on labor in liberal position was low, however, there was positive correlation between the degree of cognition and confidence in willingness to choose liberal position delivery mode ( r=0. 379, P<0. 05 ) . Relieving labor pain was the most attractive reason for pregnant women to try this delivery method. Conclusions Along with the two-child policy, enhancing the spreading and promotion of the concept of labor in liberal position, and well management the implementation of this mode of delivery method are the concerns of obstetrics health care in the future.

Objective:To investigate the role of TcpC, a virulence factor of uropathogenic Escherichia coli (UPEC), in immune evasion, and analyze its related pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 colony-forming unit of wild-type (CFT073 wt) or tcpc gene-knockout (CFT073 Δ tcpc) UPEC CFT073 strains from urethra into bladder to construct a mouse model of pyelonephritis. These mice were sacrificed 5 d after infection and their kidneys were taken to observe the gross pathological changes. Hematoxylin-eosin staining was used to observe histopathological changes in kidney tissues and immunohistochemistry was performed to locate TcpC in kidney tissues. The bacterial loads in urine samples of UPEC infected-mice were counted by ten-fold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from CFT073-infected mouse kidney or urine samples was measured by PCR. The expression of TcpC at mRNA level was detected by qRT-PCR after infecting dendritic cells with CFT073 wt strains. The influences of UPEC infection on the activation of NF-κB signaling pathway and the secretion of proinflammatory factors by dendritic cells were analyzed by Western blot and ELISA, respectively. The viability of UPEC strains in dendritic cells were observed by laser confocal microscope. Results:Compared with the CFT073 Δ tcpc group, the mice in the CFT073 wt group had obvious abscess in the kidneys as well as massive neutrophil infiltration and abundant TcpC in kidney tissues. The bacterial loads in the urine of CFT073 wt-infected mice were significantly higher than those in the urine of CFT073 Δ tcpc mice. PCR results showed that tcpc gene was successfully amplified from mouse kidney and urine samples. Increased expression of TcpC at both mRNA and protein levels was detected in CFT073 wt-infected dendritic cells. CFT073 wt infection inhibited the phosphorylation of NF-κB p50 and the production of proinflammatory factors in dendritic cells. TcpC promoted the survival of CFT073 wt in dendritic cells. Conclusions:TcpC expression increases significantly during CFT073 wt infection or in mice with CFT073 wt-induced pyelonephritis. It promotes the survival of CFT073 wt in dendritic cells by inhibiting the activation of NF-κB signaling pathway and reducing the secretion of pro-inflammatory cytokines. TcpC is involved in the pathogenesis of UPEC and immune evasion.

Background::Few evidence is available in the early prediction models of behavioral and psychological symptoms of dementia (BPSD) in Alzheimer’s disease (AD). This study aimed to develop and validate a novel genetic-clinical-radiological nomogram for evaluating BPSD in patients with AD and explore its underlying nutritional mechanism.Methods::This retrospective study included 165 patients with AD from the Chinese Imaging, Biomarkers, and Lifestyle (CIBL) cohort between June 1, 2021, and March 31, 2022. Data on demographics, neuropsychological assessments, single-nucleotide polymorphisms of AD risk genes, and regional brain volumes were collected. A multivariate logistic regression model identified BPSD-associated factors, for subsequently constructing a diagnostic nomogram. This nomogram was internally validated through 1000-bootstrap resampling and externally validated using a time-series split based on the CIBL cohort data between June 1, 2022, and February 1, 2023. Area under receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA) were used to assess the discrimination, calibration, and clinical applicability of the nomogram.Results::Factors independently associated with BPSD were: CETP rs1800775 (odds ratio [OR] = 4.137, 95% confidence interval [CI]: 1.276-13.415, P = 0.018), decreased Mini Nutritional Assessment score (OR = 0.187, 95% CI: 0.086-0.405, P <0.001), increased caregiver burden inventory score (OR = 8.993, 95% CI: 3.830-21.119, P <0.001), and decreased brain stem volume (OR = 0.006, 95% CI: 0.001-0.191, P = 0.004). These variables were incorporated into the nomogram. The area under the ROC curve was 0.925 (95% CI: 0.884-0.967, P <0.001) in the internal validation and 0.791 (95% CI: 0.686-0.895, P <0.001) in the external validation. The calibration plots showed favorable consistency between the prediction of nomogram and actual observations, and the DCA showed that the model was clinically useful in both validations. Conclusion::A novel nomogram was established and validated based on lipid metabolism-related genes, nutritional status, and brain stem volumes, which may allow patients with AD to benefit from early triage and more intensive monitoring of BPSD.Registration::Chictr.org.cn, ChiCTR2100049131.

Objective:To assess the impact of vaccination at school and influenza vaccination rates among school-age children on school absenteeism in Shenzhen.Methods:The study subjects were primary school students in Shenzhen. School absenteeism panel database from December 2017 to June 2020 of 286 primary schools in Shenzhen was merged with vaccination rates and organizational patterns ( i.e., vaccination at school vs. non-school) data of 9 districts in Shenzhen after influenza vaccination for children. The outcome was the number of school absenteeism. The treatment and control groups were distinguished by organizational patterns and district vaccination rates. Difference-in-Difference (DiD) Poisson regressions were used to analyze the effectiveness of vaccination at school and higher vaccination rates. Besides, a robustness test was performed on the regression results. Results:Poisson regression analysis and robustness test of regression results showed that vaccination at school and higher vaccination rates effectively reduced the risk of school absenteeism, with effectiveness against absenteeism of 32.6% (95% CI: 17.0%-45.3%, P<0.01) and 53.0% (95% CI: 42.1%-61.8%, P<0.01), respectively. Conclusion:A free influenza vaccination program for school-age children in Shenzhen and prioritizing school-based vaccination may be an effective measure to reduce the risk of school absenteeism.