Objective To establish the method to express leukemia inhibitory factor(LIF) in fetal fibroblasts in order to provide theoretical foundation for establishment of transgenic animal model.Methods Using the fetal of 13.5 d ICR mouse,the primary fetal fibroblasts were cultivated by trypsin enzyme digestion.The lineared eukaryotic expression vector pcDNA3.1-LIF was transferred to the fetal fibroblasts of mouse with liposome induction.The positive cells were selected by G418,genome DNA of the positive cells was extracted and sequenced. Results The primary fetal fibroblasts of mouse were successfully obtained by isolating and culturing,and fetal fibroblasts expressing LIF were established by transferring.The sequencing result demonstrated that the homology of clone plasmid of positive cells was about 99%.Conclusion Eukaryotic expression vector pcDNA3.1-LIF is successfully transferred to the fetal fibroblasts of mouse.

OBJECTIVETo establish a rapid and sensitive high-performance liquid chromatography (HPLC) method for detecting pazufloxacin concentrations in the saliva, gingival crevicular fluid and serum of healthy adults.

METHODSSamples of saliva, gingival crevicular fluid and serum were obtained from healthy adults receiving intravenous infusion of pazufloxacin. The concentrations of pazufloxacin in the samples were quantified by HPLC equipped with a reversed-phase column (Agilent Zorbax SB-C18 5 µm, 250 mm×4.6 mm). The mobile phase for pazufloxacin was a mixture of acetonitrile and 0.5% phosphoric acid containing 1% triethylamine (155:850), and 20 µl of the resulting solution was injected into the HPLC system at a flow rate of 1.0 ml/min. The detection wavelength was set at 245 nm. The samples were first deproteinized by precipitation with methanol followed by supernatant drying; the residue was reconstituted with the mobile phase and centrifuged, and the supernatants were directly injected into the HPLC system.

RESULTSPazufloxacin in the samples were totally separated without interference by any endogenous substances. The calibration curves showed a good linear regression (r>0.999). The detection limit was 10 ng/ml with within-day and between-day coefficients of variation performance all below 5% and recovery rates all above 91%.

CONCLUSIONHPLC is both sensitive and selective for quantification of pazufloxacin in saliva, gingival crevicular fluid and serum.

Adult ; Chromatography, High Pressure Liquid ; methods ; Female ; Fluoroquinolones ; analysis ; blood ; Gingival Crevicular Fluid ; chemistry ; Humans ; Male ; Oxazines ; analysis ; blood ; Saliva ; chemistry ; Sensitivity and Specificity ; Young Adult

OBJECTIVE:To prepare Bevacizumab(BEV)multivesicular liposomes(BEV-MVLs)with sustained-effect,and to study their in vitro release characteristics. METHODS:BEV-MVLs were prepared by double emulsion method. Box-Behnken design-response surface methodology was used to optimize the prescription with the concentration of glycerol trioleate(TO)in organic phase,ratio of 1,2-dioleoyl-sn-glycero-3-phosphocholine(DOPC)-cholesterol(CH)(mol/mol),the concentration of L-lysine in external water phase as factors,using encapsulation rate as index. The morphology of BEV-MVLs was observed by inverted fluorescence microscope and SEM;particle size was determined by laser particle size analyzer;the BEV content was determined by HPLC and calculate the encapsulation rate and in vitro accumulative release rate.RESULTS:The optimized prescription was as follows as TO of 2.72 mmol/L in organic phase,DOPC-CH ratio of 0.67(mol/mol)and L-lysine of 40 mmol/L in external water phase. The encapsulation rate of BEV-MVLs was(80.65±4.42)%(n=3),and relative error of it to predicted value was 2.54%. The liposomes were spherical in appearance shape and uniform in size,and they were typical non-concentric vesicle structure with average particle size of 16.80 μm. 30 d in vitro accumulative release rate was about 92%. CONCLUSIONS:Prepared BEV-MVLs show sustained-effect,and their encapsulation rate reaches the expected effect.

Morchella is a genus of fungi with the ability to concentrate Cd both in the fruit-body and mycelium. However, the molecular mechanisms conferring resistance to Cd stress in Morchella are unknown. Here, RNA-based transcriptomic sequencing was used to identify the genes and pathways involved in Cd tolerance in Morchella spongiola. 7444 differentially expressed genes (DEGs) were identified by cultivating M. spongiola in media containing 0.15, 0.90, or 1.50 mg/L Cd2+ . The DEGs were divided into six sub-clusters based on their global expression profiles. GO enrichment analysis indicated that numerous DEGs were associated with catalytic activity, cell cycle control, and the ribosome. KEGG enrichment analysis showed that the main pathways under Cd stress were MAPK signaling, oxidative phosphorylation, pyruvate metabolism, and propanoate metabolism. In addition, several DEGs encoding ion transporters, enzymaticon-enzymatic antioxidants, and transcription factors were identified. Based on these results, a preliminary gene regulatory network was firstly proposed to illustrate the molecular mechanisms of Cd detoxification in M. spongiola. These results provide valuable insights into the Cd tolerance mechanism of M. spongiola and constitute a robust foundation for further studies on detoxification mechanisms in macrofungi that could potentially lead to the development of new and improved fungal bioremediation strategies.

Morchella is a genus of fungi with the ability to concentrate Cd both in the fruit-body and mycelium. However, the molecular mechanisms conferring resistance to Cd stress in Morchella are unknown. Here, RNA-based transcriptomic sequencing was used to identify the genes and pathways involved in Cd tolerance in Morchella spongiola. 7444 differentially expressed genes (DEGs) were identified by cultivating M. spongiola in media containing 0.15, 0.90, or 1.50 mg/L Cd2+ . The DEGs were divided into six sub-clusters based on their global expression profiles. GO enrichment analysis indicated that numerous DEGs were associated with catalytic activity, cell cycle control, and the ribosome. KEGG enrichment analysis showed that the main pathways under Cd stress were MAPK signaling, oxidative phosphorylation, pyruvate metabolism, and propanoate metabolism. In addition, several DEGs encoding ion transporters, enzymaticon-enzymatic antioxidants, and transcription factors were identified. Based on these results, a preliminary gene regulatory network was firstly proposed to illustrate the molecular mechanisms of Cd detoxification in M. spongiola. These results provide valuable insights into the Cd tolerance mechanism of M. spongiola and constitute a robust foundation for further studies on detoxification mechanisms in macrofungi that could potentially lead to the development of new and improved fungal bioremediation strategies.

Objective:To explore the early clinical efficacy of ultrasound visualized platelet-rich plasma (PRP) in the treatment of lower back myofascial pain syndrome (MPS) after sports injury.Methods:A prospective cohort study was conducted to analyze the clinical data of 32 patients with lower back MPS after sports injury, who were admitted to West China Hospital of Sichuan University from January 2023 to March 2023. Ultrasound-guided PRP injection into the erector spinalis or quadratus psoas muscles was used for treatment. Before treatment, at 24 hours, 2 weeks, and 4 weeks after treatment, pain and function were evaluated using visual analogue scale (VAS), McGill pain questionnaire (McGill), Roland Morris dysfunction questionnaire (RMDQ), and Oswestry dysfunction index (ODI). Before treatment and 4 weeks after treatment, the quality of life was evaluated using the short-form 36 item health survey questionnaire (SF-36). The adverse reactions were observed during treatment and follow-up.Results:A total of 32 patients with lower back MPS after sports injury were enrolled, including 10 males and 22 females; aged 12-68 years [(47.3±16.3)years]. All the patients were followed up for 4 weeks. Before and at 24 hours, 2 weeks, and 4 weeks after treatment, the VAS was 5.0(4.0, 6.0)points, 3.5(3.0, 4.8)points, 2.0(2.0, 3.0)points, and 2.0(1.3, 3.0)points, respectively; the McGill score was 9.0(7.0, 11.0)points, 7.0(5.0, 9.0)points, 4.0(3.0, 5.0)points, and 3.0(3.0, 5.0)points, respectively; the RMDQ score was 8.0(5.3, 10.8)points, 5.5(3.0, 8.0)points, 4.0(3.0, 5.8)points, and 3.0(2.0, 4.8)points, respectively; the ODI was 22.0(14.5, 30.0), 20.0(14.5, 25.5), 9.0(6.0, 16.0), and 8.0(4.5, 14.0), respectively. Compared with the values before treatment, the VAS, McGill score, and RMDQ score were significantly decreased at 24 hours, 2 weeks, and 4 weeks after treatment (all P<0.05); the ODI had no significant difference at 24 hours after treatment ( P>0.05), but it was significantly decreased at 2 and 4 weeks after treatment (all P<0.05). Compared with the values at 24 hours after treatment, the VAS, McGill score, RMDQ score and ODI further decreased at 2 weeks after treatment (all P<0.05). Compared with the values at 2 weeks after treatment, there was no significant difference in the VAS, McGill score, RMDQ score, or ODI at 4 weeks after treatment (all P>0.05). In the SF-36, the scores of physiological function [77.5(60.0, 93.8)points], physiological role [50.0(0.0, 100.0)points], body pain [64.0(44.5, 74.0)points], vitality [75.0(65.0, 78.8)points], social function [87.5(75.0, 100.0)points], emotional role [66.7(33.3, 100.0)points] and mental health [72.0(68.0, 83.0)points] before treatment were increased to 90.0(80.0, 98.8)points, 100.0(56.3, 100.0)points, 84.0(74.0, 84.0)points, 75.0(70.0, 80.0)points, 100.0(87.5, 112.5)points, 100.0(66.7, 100.0)points, and 76.0(68.0, 84.0)points after 4 weeks of treatment, respectively ( P<0.05 or 0.01). However, there was no significant difference in the general health status or health changes before and after treatment (all P>0.05). During treatment and follow-up, no adverse reactions such as redness, swelling, pain, or subcutaneous bleeding were observed. Conclusion:Ultrasound-guided PRP treatment can improve the early pain, lumbar mobility and quality of life of patients with lower back MPS after sports injury, with no presence of adverse reactions.

Objective:To compare the efficacies of negative pressure wound therapy (NPWT) and functional dressings in primary repair of spinal cord injury complicated with lacunar soft tissue defects.Methods:A retrospective cohort study was conducted to analyze the clinical data of 30 patients with spinal cord injury complicated with lacunar soft tissue defects. The patients were admitted to West China Hospital, Sichuan University from January 2020 to December 2022, including 20 males and 10 females; aged 23-54 years [(42.1±7.8)years]. Wound site was located at the sacrococcygeal region in 16 patients, the buttock in 11, and the femoral trochanter in 3. Wound area was 28-36 cm 2 [(32.1±2.1)cm 2]. Time of wound formation was at range of 1-4 months [(2.0±0.8)months]. Among them, 15 patients received functional dressing treatment after mechanical/ultrasonic debridement (dressing treatment group), and 15 patients received NPWT treatment on the basis of mechanical/ultrasonic debridement (negative pressure treatment group). The following items were compared between the two groups: the time of primary wound repair, results of bacterial culture of wound secretions before and at the end of primary wound repair, and levels of serum interleukin-6 (IL-6) and C-reactive protein (CRP) as well as Bates-Jensen wound assessment tool (BWAT) score before, at 5 days after the primary repair and at the end of the primary repair. Results:All the patients were followed up for 3-6 months [(4.1±0.9)months]. The time of primary wound repair in the negative pressure treatment group was (13.4±2.3)days, which was markedly shorter than that in the dressing treatment group [(22.8±2.5)days] ( P<0.01). Before the primary repair, 11 patients in the negative pressure treatment group showed positive bacterial culture of wound secretions [73.3% (11/15)], and 9 patients in the dressing treatment group were positive [60.0% (9/15)] ( P>0.05). At the end of primary repair, there was 1 patient with positive bacterial culture of wound secretions in the negative pressure treatment group [6.7% (1/15)], which was less than 7 patients in the dressing treatment group [46.7% (7/15)] ( P<0.05). The numbers of positive patients at the end of the primary repair were lower than those before the primary repair in both groups, and the difference in the negative pressure treatment group was statistically significant ( P<0.01), with no significant difference found in the dressing treatment group ( P>0.05). Before the primary repair, the IL-6, CRP and BWAT score were 20.5(8.4, 32.3)pg/ml, 24.2(14.7, 33.0)mg/L, and (37.1±4.8)points in the negative pressure treatment group, comparable with 13.8(11.8, 35.4)pg/ml, 23.6(13.1, 52.3)mg/L, and (35.2±4.7)points in the dressing treatment group (all P>0.05). At 5 days after primary repair, the IL-6, CRP and BWAT score in the negative pressure treatment group were 20.2(7.9, 28.6)pg/ml, 20.0(11.6, 30.5)mg/L, and (34.9±4.3)points, comparable with 11.6(8.9, 20.6)pg/ml, 25.3(10.0, 50.3)mg/L, and (35.2±4.5)points in the dressing treatment group (all P>0.05). At the end of primary repair, the IL-6, CRP and BWAT score were 2.3(1.5, 4.5)pg/ml, 4.8(3.7, 6.9)mg/L, and (23.6±1.8)points in the negative pressure treatment group, statistically different from 4.4(3.3, 6.9)pg/ml, 8.4(5.5, 31.4)mg/L, and (31.4±3.3)points in the dressing treatment group (all P<0.01). The IL-6, CRP and BWAT score at the end of the primary repair were significantly different compared with those before and at 5 days after the primary repair in the two groups ( P<0.05 or 0.01). However, no significant difference was found between the two groups before and at 5 days after the primary repair (all P>0.05). Conclusion:Compared with functional dressings, NPWT can shorten the time required for primary repair of spinal cord injury complicated with lacunar soft tissue defects, control the inflammatory state of the wound, improve the trend of wound healing, and create a good condition for secondary repair treatment of the wound.

The problems caused by proximal contact loss (PCL) of dental implants have been a mainstream research topic in recent years, and scholars are unanimously committed to analyzing their causes and related factors, aiming to identify solutions to the problems related to PCL. The effects of the anterior component of force (ACF), the lifelong remolding of the adult craniofacial jaw and alveolar socket, and the osseointegration characteristics of dental implants are the main causes of PCL. On the one hand, the closing movement of the mandible causes the ACF of the tooth to move through the posterior molar cusp. Moreover, drifting between the upper and lower posterior teeth and mandibular anterior teeth can cause the anterior teeth of the upper and lower jaws to be displaced labially. On the other hand, reconstruction of the jaw, alveolar socket and tooth root, the forward horizontal force of the masticatory muscles, the dynamic component of the jaw and the forward force generated by the oblique plane of the tooth cusp can cause the natural tooth to experience near-middle drift. Additionally, natural teeth can shift horizontally and vertically and rotate to accommodate remodeling of the stomatognathic system and maintain oral function. Nevertheless, the lack of a natural periodontal membrane during implant osseointegration, the lack of a physiological basis for near-medium drift, the small average degree of vertical motion and the integrated silence of dental implants without the overall drift characteristics of natural teeth increases the probability of PCL. The high incidence of PCL is clearly associated with the duration of prosthesis delivery and the mesial position; but it is also affected by the magnitude of the bite force, occlusion, the adjacent teeth, restoration design, implant location, jaw, and patient age and sex. PCL has shown a significant correlation with food impaction, but not a one-to-one correspondence, and did not meet the necessary and sufficient conditions. PCL is also associated with peri-implant lesions as well as dental caries. PCL prevention included informed consent, regular examinations, selection of retention options, point of contact enhancement, occlusal splints, and the application of multipurpose digital crowns. Management of the PCL includes adjacent contact point additions, orthodontic traction, and occlusal adjustment. Existing methods can solve the problem of food impaction in the short term with comprehensive intervention to seek stable, long-term effects. Symmetric and balanced considerations will expand the treatment of issues caused by PCL.