BACKGROUND:The micro/nanostructured gradient biomimetic surface of implant materials can simulate the structure of the extracellular environment in human bone tissue,thereby achieving perfect bone integration function.However,further research is needed on the mechanisms by which the surface microstructure of bone implant materials regulates cell function and promotes osteogenesis. OBJECTIVE:To analyze the effect of titanium sheet microstructure surface on osteogenic differentiation of MC3T3-E1 osteoblasts. METHODS:(1)At a constant voltage of 5 V or 20 V,nanotube arrays of different diameters were prepared on the surface of titanium sheets by acid etching and anodic oxidation techniques,and were recorded as group R5 and group R20,respectively.The surface morphology,roughness,and hydrophilicity of pure titanium sheet(without acid etching or anodizing treatment)were measured in group R5 and group R20.(2)MC3T3-E1 osteoblasts of logarithmic growth stage were inoculated on the surface of pure titanium sheets,R5 group and R20 group respectively.After 24 hours of osteogenic induction culture,the expression of mechanical sensitive channel protein 1 was analyzed by RT-PCR and immunofluorescence staining.Osteoblast inducible base with or without the mechanosensitive channel protein 1 activator Yada1 was added,and alkaline phosphatase staining was performed after 7 days of culture.Alizarin red staining was performed after 14 days of culture. RESULTS AND CONCLUSION:(1)The surface of pure titanium sheets was smooth under scanning electron microscope.Relatively uniform and orderly nanotube arrays with average diameters of about 30 nm and 100 nm were observed on the surface of titanium sheets of groups R5 and R20,respectively.The results of scanning electron microscope were further verified by atomic force microscopy.The surface roughness of titanium sheet of group R5 was higher than that of pure titanium(P<0.05),and the water contact angle was lower than that of pure titanium(P<0.05).The surface roughness of titanium sheet in group R20 was higher than that in group R5(P<0.05),and the water contact angle was lower than that in group R5(P<0.05).(2)RT-PCR and immunofluorescence staining showed that the expression of mechanosensitive channel protein 1 in group R5 was higher than that in pure titanium group(P<0.05),and the expression of mechanosensitive channel protein 1 in group R20 was higher than that in group R5(P<0.05).Under the osteogenic induction,compared with the condition without Yada1,there were no significant changes in the activity of alkaline phosphatase and the deposition of calcified nodules in pure titanium group after Yada1 addition,while the activity of alkaline phosphatase and the deposition of calcified nodules in groups R5 and R20 after Yada1 addition were significantly increased(P<0.05).With or without Yada1,the alkaline phosphatase activity and calcified nodule deposition in group R5 were higher than those in pure titanium group(P<0.05),and the alkaline phosphatase activity and calcified nodule deposition in group R20 were higher than those in group R5(P<0.05).(3)The results show that the surface microstructure of titanium sheet can promote the osteogenic differentiation of osteoblast MC3T3-E1 by activating mechanosensitive channel protein 1.

Objective:To analyze the potential clinical biological factors influencing the major pathological response (MPR) to neoadjuvant immunochemotherapy in patients with resectable head and neck squamous cell carcinoma (HNSCC).Methods:This retrospective study enrolled patients with resectable HNSCC who underwent neoadjuvant immunochemotherapy at Sun Yat-sen University Cancer Center from June 1, 2019 to December 31, 2021. Binary logistic regression was used to analyze the correlation between clinical characteristics, inflammatory markers and MPR, and a nomogram model was constructed. The calibration curve and decision curve analysis were used to verify the predictive ability and accuracy of the nomogram model.Results:A total of 173 patients were included in the study, with 141 males and 32 females, aged from 22 to 83 years. After pathological assessment, the patients were divided into two groups: MPR group (108 cases) and non MPR group (65 cases). Logistics regression analysis indicated that the patients with HPV+oropharyngeal cancer, partial response or complete response by imaging assessment, low pre-treatment platelet/lymphocyte ratio, low pre-treatment C reactive protein/albumin ratio and lower pre-and post-treatment C reactive protein/albumin ratio difference were more likely to have MPR (all P<0.05). Nomogram model was constructed based on the above factors, with a C-index of 0.826 (95% CI: 0.760-0.892), and the calibration curve and decision curve analysis confirmed the prediction accuracy of the model. Conclusion:This study shows that many factors are related to MPR of patients with resectable HNSCC receiving neoadjuvant immunochemotherapy and the constructed nomogram model helps to develop personalized treatment strategies for the patients.

Objective To investigate the effect of dexmedetomidine on renal function in patients with septic-associated acute kidney injury(SAKI).Methods A prospective cohort study was conducted in 180 patients with sepsis admitted to ICU in the Second Affiliated Hospital of Guangdong Medical University from October 2021 to April 2023.According to the principle of randomized controlled trials,60 non-acute kidney injury(AKI)patients were divided into S-D group(n = 30,dexmedetomidine + conventional treatment)and S group(n = 30,conventional treatment),and the occurrence and disease score of AKI after treatment were compared between the two groups.A total of 120 AKI patients were divided into SA-D-RT group(n = 30,dexmedetomidine + blood purification + conven-tional treatment),SA-D group(n = 30,dexmedetomidine + conventional treatment),SA-RT group(n = 30,blood purification + conventional treatment)and SA group(n = 30,conventional treatment).Renal function,inflammatory factor level and disease change of the four groups were compared after treatment.Results After treatment,the inci-dence of AKI in S-D group was lower than that in S group,and the APACHEII score and SOFA score in S-D group were lower than those in S group on the 7th day after treatment(P<0.05).After 7 days of treatment,the level of Scr,BUN and CysC in the 4 groups was significantly lower than that on the 1st and 3rd day,and those in the SA-D-RT group were lower than those in the SA-D group,SA-RT group and SA group(P<0.05).After 7 days of treatment,the level of CRP,PCT,TNF-α,IL-6 and IL-1β in four groups was significantly decreased compared with on the 1st and 3rd day,and the level of CRP,PCT,TNF-α,IL-6 and IL-1β in SA-D-RT group was lower than that in SA-D group,SA-RT group and SA group(P<0.05).After 7 days of treatment,the APACHEII score and SOFA score of the four groups were significantly lower than on the 1st and 3rd day,and the scores of the SA-D group were lower than those of the SA-D group,the SA-RT group and the SA group(P<0.05).Conclusion Dexmedetomidine can effectively reduce the incidence of AKI,affect the expression level of renal function markers and inflammatory factors in serum of SAKI patients,and improve the condition of patients.

Objective:To investigate the genetic safety of allogeneic bone marrow mesenchymal stem cells (BMSCs) transplantation in traumatic brain injury (TBI).Methods:(1) In vivo experiment: BMSCs from male SD rats were isolated and cultured. Moderate TBI models were prepared by implanting and fixing micro-drug injection cannula into the left ventricle of 12 female SD rats, and 3 d after that, striking the right cerebral cortex of the rats with pneumatic precision percussion device was performed. Four h, and 3, 6, 9, and 12 d after modeling, TBI rats were given a single/multiple BMSCs infusion (2.5×10 5/time, total volume 10 μL) by cannula; 48 and 72 h, and 10 and 14 d after modeling, brain tissues of TBI rats (3 at each time point) were prepared into paraffin specimens. Immunofluorescent staining was used to detect the microglia activation, and RNAscope ? technology was used to detect the co-localization of astrocytes, neurons, microglia and transplanted BMSCs to observe whether the allogeneic BMSCs were integrated with the host brain cells after transplantation into TBI host. (2) In vitro experiment: the frozen and revived microglial cell line BV2 was transfected with green fluorescent protein (GFP)-positive lentiviral particles, and then, BMSCs prelabeled with pHrodo RED probe and BV2 cells pretreated with lipopolysaccharide were co-cultured in a certain ratio (BV2:BMSCs=1:1, 1:2, 2:1); after 36 and 72 h of co-culture, the phagocytosis between the 2 kinds of cells was observed under confocal fluorescence inverted microscope to observe the specific action forms of microglia on BMSCs. Results:(1) In vivo experiment: 48 and 72 h, and 10 and 14 d after modeling, no colocalization of transplanted BMSCs with astrocytes or neurons was found in paraffin sections of brain tissue in TBI rats; however, 10 and 14 d after modeling, microglia in TBI rats were obviously activated and migrated to the left lateral ventricle and choroid plexus, and co-localization of microglia with transplanted BMSCs was observed. (2) In vitro experiment: phagocytosis occurred after co-culture of BV2 cells at different proportions with BMSCs for 36 and 72 h. Conclusion:After transplantation, allogeneic BMSCs do not integrate with astrocytes or neurons of the TBI host, but they could be phagocytosed by microglia, indicating that allogeneic BMSCs transplantation for TBI is genetically safe.

Objective: To explore the effects of long noncoding-RNA (lncRNA) taurine upregulated gene 1 (TUG1) on the proliferation and osteogenic/odontoblast differentiation of human dental pulp stem cells (hDPSCs). Methods : hDPSCs were isolated and cultured. The surface antigens CD44, CD45, CD73, CD90, CD133 and STRO-1 were detected by flow cytometry. Alkaline phosphatase (ALP) staining and alizarin red staining were used to identify the ability of cells to differentiate. RNA was collected on Days 0, 7 and 14 of the osteogenic induction of hDPSCs, and qRT-PCR was used to detect the relative expression of TUG1. The hDPSCs were stably transfected with a lentiviral vector containing the TUG1-silenced pSLenti-U6-shRNA(TUG1)-CMV-EGFP-F2A-Puro-WPRE to silence TUG1. The ability of hDPSCs to proliferate was assessed with the CCK-8 method. ALP and alizarin red staining and quantitative detection were used to detect the ALP activity and formation of mineralized nodules of hDPSCs. The expression levels of dentin sialophosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), Runt-associated transcription factor 2 (Runx2), osteocalcin (OCN) and osteopontin (OPN) genes and proteins were measured by qRT-PCR and Western blot. Results : The hDPSCs were successfully isolated and cultured, and TUG1 expression was significantly increased during osteogenic differentiation (P<0.05). The hDPSCs proliferation was suppressed after silencing TUG1(P<0.05). After osteogenic induction, ALP and alizarin red staining showed that ALP activity and mineralized nodules were suppressed by silencing TUG1. The expression levels of the odontogenic differentiation gene DSPP and DMP-1 and the osteogenic differentiation gene Runx2, OCN and OPN were also significantly decreased (P<0.05). Conclusion Knocking down TUG1 can inhibit the proliferation and osteogenic/odontogenic differentiation of hDPSCs.

Objective:To explore the anatomical and clinical effects of lateral-perineal approach in treating the fracture of inferior ramus of pubis-ischium ramus.Methods:The lateral approach of the perineum was simulated on 10 side of 5 intact wet adult cadavers to determine the surface symbols of incision design and to expose the operative field of the approach and to observe the anatomical characteristics of the perineal branch of the posterior femoral cutaneous nerve and the incision of the surgical approach. Five points were selected at the incision of the approach. The distance (L 1-L 5) between each point and the body surface projection of the perineal branch of the posterior femoral cutaneous nerve was measured. A total of 11 patients, including 5 males and 6 females with an average age of 41.55±14.32 years, ranging from 18 to 62 years, were treated by this approach in clinical practice. All patients had a reduction and fixation to the fracture of inferior ramus of pubis-ischium ramus. The operation duration, incision length, intraoperative blood loss and surgical complications were recorded. The quality of fracture reduction was evaluated according to Matta radiographic criteria. The strength of the adductor was measured. The pelvic function was evaluated according to Majeed Pelvic Score at the last follow-up. Results:Anatomical studies shown that the line between the two points. One point was 4 cm lateral to the level of the apex of the pubic arch. Another point was 4 cm from the ischial tubercle on the line from the ischial tuberosity to the point that 4 cm lateral to the level of the apex of the pubic arch was the axis of the approach lateral of the perineum. Anatomical studies showed that the lateral-perineal approach could expose the range from pubic symphysis to sciatic tuberculum. The distance between the points selected at the incision of the approach and the body surface projection of the perineal branch of the posterior femoral cutaneous nerve: L 1 was 19.40±1.17 mm, ranging from 18 to 21 mm; L 2 was 16.60±2.76 mm, ranging from 10 to 20 mm; L 3 was 18.30±1.89 mm, ranging from 16 to 21 mm; L 4 was 19.20±1.93 mm, ranging from 16 to 22 mm; L 5 was 14.70±1.83 mm, ranging from 13 to 18 mm. All patients were followed up for 17.91±4.09 months, ranging from 13 to 26 months. The incision length was 8.18±0.98 cm, ranging from 7 to 10 cm. The operation duration was 59.64±12.17 min, ranging from 43 to 85 min. The intraoperative blood loss was 100 ml, ranging from 50 to 130 ml. All incisions were healed in all patients. The fractures were healed in 13.36±2.06 weeks, ranging from 10 to 16 weeks. According to Matta radiographic criteria, the quality of fracture reduction was excellent in 6 cases, good in 4 cases and fair in 1 case. At the last follow-up, the adductor muscle strength reached grade 4 in 4 patients and grade 5 in 7 patients. Furthermore, according to the Majeed Pelvic Score, the score of every patient was 86.55±9.59, ranging from 66 to 100, and 8 cases were excellent, 3 cases were good at the last follow-up. The heterotopic ossification occurred in 2 patients, the pain during intercourse occurred in 2 patients. No patient had sensory disturbance or pain in the perineal area. Conclusion:A certain safe distance is between the lateral to the perineum and the perineal branch of the posterior femoral cutaneous nerve with limited risk of injuring posterior femoral cutaneous nerve via the lateral approach of the perineum. The advantages in treating the fracture of inferior ramus of pubis-ischium ramus by this approach have concealed incision, short operation duration and less bleeding with satisfied short-term clinical effects.

Objective: To investigate the inhibitory effect of honeysuckle on Streptococcus mutans UA159 in vitro. Methods: We used a double-dilution method to measure the minimum inhibitory concentration (MIC) of honeysuckle against Streptococcus mutans UA159. Lonicerae lonicerae powder was dissolved in the solvent DMSO, different concentrations of liquid medicine were prepared, and bacterial liquid was added. The solution control group and bacterial liquid control group were set at the same time. The growth and acid production of UA159 were determined using antibacterial experiments. A growth curve and acid production curves were drawn, and the adhesion rate and adhesion inhibition rate were calculated. The effect of honeysuckle on the formation of Streptococcus mutans UA159 was determined by crystal violet quantification, and a microscope and a scanning electron microscope were used to observe biofilm formation and structural changes. Results : The MIC of honeysuckle against Streptococcus mutans UA159 was 12.5 mg/mL. The bacteriostatic experiments showed a difference in the growth, acid production and adhesion of UA159 after honeysuckle treatment (P<0.05) compared with the controls, and the inhibitory effect increased as the drug liquid concentration increased. Crystal violet quantification showed a significant difference in biofilm formation between the pharmaceutical liquid group and the control group (P<0.05). Meanwhile, the forward microscope showed a significant decrease in biofilm formation. Under SEM, the number of bacteria decreased significantly at 0, 6 and 12 h after honeysuckle addition. Conclusion Honeysuckle inhibits the growth and acid production of UA159 and inhibits adhesion and the formation of biofilms.

Objective:To develop a model of stellate ganglion block (SGB) in mice and investigate the effect of SGB on cerebral cortical blood flow.Methods:Thirty clean-grade healthy male C57BL/6 mice, aged 8-9 weeks, weighing 23-27 g, were divided into 5 groups ( n=6 each) using a random number table method: control group (group C), left SGB group (group L), left normal saline group (group SL), right SGB group (group R) and right normal saline group (group SR). Group C received no intervention.SGB was performed with 0.25% ropivacaine 0.08 ml via percutaneous posterior approach in L and R groups, while the equal volume of normal saline 0.08 ml was given instead at the location of left and right stellate ganglion in SL and SR groups, respectively.The cerebral cortical blood flow was determined using laser speckle contrast imaging system before SGB (T 0) and at 10, 30, 60, 90 and 120 min after SGB (T 1-5). Results:Mice developed ptosis on the block side, indicating that the model of SGB was successfully developed in L and R groups.There was no significant difference in cerebral cortical blood flow at each time point among C, SL and SR groups ( P>0.05), and cerebral cortical blood flow on the block side decreased at T 1, began to increase at T 2, peaked at T 3, and decreased at T 5 which was still higher than that at T 0 in group L and group R ( P<0.01). Compared with C and SL groups, the left cerebral cortical blood flow was significantly decreased at T 1, 5 and increased at T 2-4 in group L ( P<0.01). Compared with C and SR groups, the right cerebral cortical blood flow was significantly decreased at T 1, 5 and increased at T 2-4 in group R ( P<0.01). There were no significant differences in cerebral cortical blood flow at each time point between group C and group SL and between group C and group RL ( P>0.05). Conclusions:The mouse model of SGB via percutaneous posterior approach is successfully developed.Unilateral SGB can affect cerebral cortical blood flow on the block side, which shows a transitory decrease followed by a sustained significant increase.

Background: Excessive delivery of free fatty acids (FFAs) to the liver promotes steatosis and insulin resistance (IR), with IR defined as reduced glucose uptake, glycogen synthesis and anti-lipolysis stimulated by normal insulin levels. Whether the associations between FFAs and diabetes development differ between patients with and without nonalcoholic fatty liver disease (NAFLD) remains unclear. Methods: Consecutive subjects (2,220 NAFLD subjects and 1,790 non-NAFLD subjects according to ultrasound imaging) were enrolled from the First Affiliated Hospital of Sun Yat-sen University between 2009 and 2019. The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. Results: There was an approximate J-shaped relationship between FFA levels and HOMA-IR in the NAFLD group. Higher FFA concentration quartiles were associated with higher risks of IR (odds ratio [OR], 9.24; 95% confidence interval [CI], 6.43 to 13.36), prediabetes (OR, 10.48; 95% CI, 5.66 to 19.39), and type 2 diabetes mellitus (T2DM; OR, 19.43; 95% CI, 12.75 to 29.81) in the NAFLD group but not in the non-NAFLD group. The cut-off points for the FFA levels increased in a stepwise manner in discriminating IR, prediabetes and T2DM (573, 697, and 715 μmol/L) in the NAFLD group but not in non-NAFLD individuals. Conclusion A distinct dose-dependent relationship of FFA levels was found with IR, prediabetes and T2DM in NAFLD patients. Screening serum FFA levels in NAFLD patients would be valuable in preventing diabetes development.

Objective:To investigate the clinical efficacy of anterior approach combined with blocking plates and screws in the management of acetabular fracture involving the quadrilateral area.Methods:A retrospective case series analysis was performed for 16 patients with acetabular fracture involving the quadrilateral area admitted to First and Second Affiliated Hospital of Guangxi Medical University from January 2017 to January 2019. There were 12 males and 4 females，with the age of 21-66 years［（45.3±10.6）years］. According to Letournel-Judet classification，there were 9 patients with bi-column fracture，6 with anterior and posterior traverse fracture and 1 with anterior column fracture. A total of 9 patients were operated via the ilioinguinal approach and 7 via the lateral-rectus approach. Reduction and fixation of the pelvis and acetabulum were performed，using 3.5 mm cortical bone screws or plates to block the internal displacement of fracture in the quadrilateral body. The incision length，operation time and intraoperative blood loss were recorded. The quality of fracture reduction was assessed according to the Matta reduction criteria at postoperative 2 days and hip function by the modified Merle D'Aubigne-Postel score at postoperative 3 months and 12 months. Postoperative complications were observed.Results:All patients were followed up for 13-24 months［（16.1±2.9）months］. The ilioinguinal approach and lateral-rectus approach showed surgical incision of 12-26 cm［（18.6±4.0）cm］and 8-15 cm［（10.7±2.3）cm］，respectively. The operation time was 107-215 minutes［（159.2±27.8）minutes］and the intraoperative blood loss was 200-2，300 ml［（853.1±489.7）ml］. According to Matta reduction criteria，the results were excellent in 9 patients and good in 7. Three months after operation，the modified Merle D'Aubigne-Postel score was 11-18 points［（15.2±2.2）points］，which showed the results were excellent in 4 patients，good in 7，fair in 4 and poor in 1，with the excellent and good rate of 69%. Twelve months after operation，the modified Merle D'Aubigne-Postel score was 13-18 points［（16.9±1.4）points］，which showed the results were excellent in 7 patients，good in 8 and fair in 1，with the excellent and good rate of 94%. The liquefaction of post-surgical incision was seen in a patient，bladder injury in a patient，lateral femoral cutaneous nerve injury in a patient，and heterotopic ossification in a patient. There was no loosening or breakage of the internal fixation.Conclusion:For acetabular fracture involving the quadrilateral area，anterior approach combined with blocking plates and screws can prevent the displacement of quadrilateral fracture and attain satisfactory reductiongood hip function recovery and few complications.