BACKGROUND:The implant osseointegration rate of patients with diabetes is low,and the failure rate is high,which seriously affects the quality of life.It is urgent to improve the implant osseointegration of patients with diabetes by effective means to elevate the success rate.Exploring the effect of berberine on the osteogenic differentiation of bone marrow mesenchymal stem cells under a high-glucose environment and its specific mechanism will provide effective theoretical support for solving the above problems. OBJECTIVE:To explore the effect of natural extract berberine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells under the high-glucose microenvironment. METHODS:Bone marrow mesenchymal stem cells of SD rats were cultured by the whole bone marrow adherence method.CCK-8 assay was used to detect the effects of different concentrations of berberine on the proliferation of bone marrow mesenchymal stem cells under the high-glucose environment and to screen out the optimal berberine concentration.The expressions of Runx2 and Osx were detected by alkaline phosphatase activity,alicarin red staining and PCR to determine the effect of berberine on osteogenic differentiation of bone marrow mesymal stem cells under the high-glucose environment.To further explore the underlying mechanism,we introduced the AMPK-specific inhibitor Dorsomorphin and used a DCFH-DA reactive oxygen species fluorescent probe to examine reactive oxygen species levels.The p-AMPK expression was also determined by western blot assay. RESULTS AND CONCLUSION:(1)10 μmol/L was the optimal concentration of berberine to promote bone marrow mesenchymal stem cell proliferation.(2)Alberberine promoted alkaline phosphatase viability of bone marrow mesenchymal stem cells and mineralized nodule formation in a high-glucose microenvironment.(3)Alberberine promoted the expression of Runx2 and OSx in a high-glucose microenvironment.(4)Alberensine effectively inhibited the reactive oxygen species level of bone marrow mesenchymal stem cells in a high-glucose environment.(5)The effects of berberine on promoting bone marrow mesenchymal stem cell osteogenesis and inhibition of reactive oxygen species were reversed by the AMPK inhibitor.(6)Berberine activated AMPK and promoted p-AMPK expression.(7)The above results indicate that berberine(10 μmol/L)promotes the osteogenic differentiation of bone marrow mesenchymal stem cells in a high-glucose environment by activating AMPK and reducing intracellular reactive oxygen species levels.

Objective To evaluate the applicability of a modified U-shaped forearm flap for the repair of small-and medium-sized defects in the oral and maxillary areas to provide a reference for clinicians.Methods This study was re-viewed and approved by the Ethics Committee,and informed consent was obtained from the patients.Ten patients with small-and medium-sized defects in the oral and maxillary areas underwent surgical repair using modified U-shaped fore-arm flaps.There were 8 males and 2 females aged 43-72 years.The donor site was apposed primarily after harvesting the modified U-shaped forearm skin flap.The flaps ranged from 6 cm × 4 cm to 8 cm × 5 cm in size.Six months after the operation,hand movements(finger extension,fist clenching,wrist rotation upward and wrist rotation downward),the forearm donor site,hand sensations and the satisfaction score for the postoperative quality of the scar at the donor site were evaluated(0 to 10;0:very unattractive,10:very satisfactory).Results A total of 10 patients with modified U-shaped forearm flaps survived.One patient developed venous crisis 24 hours after surgery and survived after surgical ex-ploration.Delayed healing occurred at the donor site of the forearm in 1 patient,and the wounds at the donor site of the forearm in the other patients all healed in the first stage.One patient presented with dysesthesia in the hand 2 weeks af-ter surgery and recovered within 3 months.Six months after surgery,all patients had no limited hand movement and no paresthesia at the forearm donor site or hand.The patients were basically satisfied with the appearance of the donor site,and the average satisfaction score of the subjective questionnaire was 8.4 points.Conclusion Modified U-shaped fore-arm flaps can directly close forearm donor site wounds,which avoids surgical trauma to the secondary donor site and sig-nificantly reduces related complications.Modified U-shaped forearm flaps provide an alternative to conventional forearm flaps for the repair of small-and medium-sized defects in the oral and maxillary areas.

Objective To evaluate the protective effect and underlying mechanism of ulinastatin on hepatic ischemia-reperfusion injury.Methods Twenty-four male SD rats were divided into three groups:sham operation group(Sham group),hepatic ischemia-reperfusion injury group(HIRI group)and hepatic ischemia-reperfusion injury+ulinastatin pretreatment group(HIRI+UTI group),with 8 rats in each group.The HIRI rat models were established by occluding hepatic portal vein and hepatic artery for 1 h.In the HIRI+UTI group,the rats were intraperitoneally injected with ulinastatin at 30 min before model establishment,and an equivalent amount of normal saline was given in the Sham and HIRI groups.The rats were sacrificed at 6 h after model establishment.Serum samples were collected to detect alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.Ultrastructural changes of mitochondria in liver tissues were observed by transmission electron microscopy.The expression of glutathione peroxidase 4(GPX4)was determined by immunofluorescent staining.The contents of malondialdehyde(MDA),glutathione(GSH),Fe,reactive oxygen species(ROS)and GPX4 were detected.The expression levels of GPX4 and acyl-CoA synthetase long-chain family 4(ACSL4)messenger RNA(mRNA)and proteins in liver tissue were measured.Results Compared with the Sham group,serum ALT and AST levels were up-regulated,pathological changes such as congestion,hepatocyte necrosis and abnormal hepatic lobule structure were observed,pathological score was increased,the mitochondria shrank,the membrane density was increased,the mitochondrial crest was damaged or even absent,the contents of ROS,MDA and Fe were elevated,the GSH content was decreased,the fluorescent intensity of GPX4 was weakened,the relative expression levels of ACSL4 mRNA and protein were up-regulated,and the relative expression levels of GPX4 mRNA and protein were down-regulated in the HIRI group(all P＜0.05).Compared with the HIRI group,serum ALT and AST levels were down-regulated,liver tissue injury was alleviated,pathological score was decreased,mitochondrial shrinkage and crest breakage were mitigated,the contents of ROS,MDA and Fe were down-regulated,the GSH content was up-regulated,the fluorescent intensity of GPX4 was enhanced,the relative expression levels of ACSL4 mRNA and protein were down-regulated,and the relative expression levels of GPX4 mRNA and protein were up-regulated in the HIRI+UTI group(all P＜0.05).Conclusions Ulinastatin may alleviate hepatic ischemia-reperfusion injury in rats probably through inhibiting ferroptosis.

Objective To investigate the effect of sigma-1 receptor(sig-1R)inhibition on apoptosis in the dorsal root ganglion(DRG)of rats with sciatic nerve chronic constriction injury(CCI)-mediated neuropathic pain.Methods Rats undergoing intrathecal intubation were randomly divided into three groups with 12 in each:sham group,model group(CCI modeling one week after intrathecal intubation)and sig-1R inhibitor group(BD1047 group)that was injected intrathecally on the fourth,fifth,and sixth days after the CCI operation.The mechanical withdrawal threshold(MWT)of rats was detected on the day before surgery and then first,third,fifth and sev-enth days after surgery.The expression of sig-1R,Bcl-2 and Bax was detected by Western blot and immunofluo-rescence in DRG cell;TUNEL staining microscopy was used to observe the apoptosis of DRG cells.The changes of endoplasmic reticulum and mitochondria were observed by transmission electron microscopy of DRG cells.Results Compared with the sham group,the model group showed a decreased MWT at all time points after surgery,up-regulation of DRG cell apoptosis,up-regulation of sig-1R and Bax,down-regulation of Bcl-2,swelling of en-doplasmic reticulum,disruption of mitochondrial membrane and reduction of mitochondrial cristae in the DRG cell endoplasmic reticulum after surgery(P＜0.05).BD1047 group showed elevated MWT at the fifth and seventh postoperative days,down-regulation of DRG cell apoptosis,down-regulated expression of sig-1R and Box,up-regulated expression of Bcl-2 and slightly damaged endoplasmic reticulum as well as mitochondria of DRG cells compared with model group(P＜0.05).Conclusions Inhibition of sig-1R up-regulates mechanical withdrawal threshold and reduces DRG cell apoptosis in CCI rats.

Objective:To evaluate the relationship between immunoglobulin heavy chain-binding protein (BIP) and Na v1.8 in peripheral nerve in a rat model of neuropathic pain. Methods:Forty-four SPF healthy male Sprague-Dawley rats, weighing 210-260 g, were used in this study.Neuropathic pain was induced by chronic constriction injury (CCI) in anesthetized rats.The experiment was performed in two parts.Experiment Ⅰ Twenty rats were divided into 2 groups ( n=10 each) using a random number table method: sham operation group (group Sham) and group CCI.Experiment Ⅱ Twenty-four rats were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (group Sham), CCI plus normal saline group (group CCI+ NS) and CCI plus BIP inhibitor HA15 group (group CCI+ H). Starting from 4th day after surgery, 0.9% normal saline 1 ml was intraperitoneally injected in group CCI+ NS, and HA15 0.7 mg/kg was intraperitoneally injected in group H, once a day for 3 consecutive days.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before surgery and 3, 5 and 7 days after surgery (T 0-T 4), and the expression of BIP and Na v1.8 in dorsal root ganglion (DRG) and sciatic nerve was detected by Western blot on 7th day after completion of behavioral testing in two groups.The expression and colocalization of BIP and Na v1.8 in DRG and sciatic nerve were determined by immunofluorescence on 7th day after completion of behavioral testing in group Ⅰ, and the interaction between BIP and Na v1.8 was evaluated by co-immuno-precipitation. Results:Experiment Ⅰ Compared with group Sham, the MWT was significantly decreased, and TWL was shortened at T 1-T 4, the expression of Na v1.8 in DRG was down-regulated, the expression of BIP was up-regulated, and the expression of Na v1.8 and BIP in sciatic nerve was up-regulated in group CCI ( P<0.05), and BIP and Na v1.8 on the sciatic nerve were co-localized, BIP could co-precipitate Na v1.8 from DRG, and Na v1.8 could also coprecipitate BIP in group CCI.Experiment Ⅱ Compared with group Sham, the MWT was significantly decreased, and TWL was shortened at T 1-T 4, the expression of Na v1.8 in DRG was down-regulated, the expression of BIP was up-regulated, and the expression of Na v1.8 and BIP in sciatic nerve was up-regulated in group CCI+ NS ( P<0.05). Compared with group CCI+ NS, the MWT was significantly increased, and TWL was prolonged at T 3, 4, and the expression of Na v1.8 in DRG was down-regulated in group CCI+ H ( P<0.05). Conclusion:BIP can mediate the redistribution of Na v1.8 in peripheral nerve and is involved in the pathophysiological mechanism of neuropathic pain in rats.

Objective By observing the incidence of postoperative hyperalgesia in patients with total intravenous anesthesia,to study the effects of different anesthesia depth on postoperative hyperalgesia.Methods Forty gynecologic patients undergoing open lower abdominal operation were randomized into two groups (n=20): group B1 with BIS 40-49 and the group B2 with BIS 50-59.Anaesthesia was induced and maintained with intravenous anesthetics.The threshold of haphalgesia and the range of mechanical hyperalgesia were measured before operation.The consumption of the intravenous anesthetics,the emergence time and the extubation time were recorded.The vital signs were recorded at eight time points: pre-aneasthesia (T0),abdominal exploration (T1),the end of operation (T2)and 1 h (T3),4 h (T4),12 h (T5),24 h (T6),48 h (T7) after extubation.The VAS scores at T3-T7 were recorded.The PCA dosage of analgesic and superaddition in postoperative 24 h and 48 h were recored.The Von Frey Hairs were used to assessed the threshold of haphalgesia and the range of mechanical hyperalgesia at T6 and T7.Results The MAP at T6 and T7 and the VAS scores at T6 of the group B1 were lower than those of the group B2 (P<0.05).The range of mechanical hyperalgesia at T7 of the group B1 was narrower than that of the group B2 (P<0.05).Conclusion Maintaining the anesthesia depth within BIS 40-49 can reduce the body′s stress reaction,decrease the incidence of postoperative hyperalgesia and accelerate the recovery time of mechanical hyperalgesia in postoperative 48 h.

BACKGROUND: Apical sealing ability is the key to ensure the long-term curative effect of root canal therapy. The post space preparation exposes some inevitable influence on root canal sealing ability, so how to minimize this effect becomes a hot spot.OBJECTIVE: To explore the effects of immediate or delayed post space preparation on the apical sealing ability of different root canal sealers.METHODS: Forty-eight extracted human premolar teeth were obtained, and the tooth crown was cut off. The samples were randomly divided into three groups (n=16 teeth per group). Group A underwent the immediate post space preparation; group B underwent the delayed post space preparation; group C without the post space preparation. Then all groups were subdivided into two groups, and then were filled with the gutta-percha/AH-Plus (groups A1, B1 and C 1)or the gutta-percha/mineral trioxide aggregate (groups A2, B2 and C2). The depth of apical dye penetration was measured using pressure-driven system. RESULTS AND CONCLUSION: There were no significant differences in the apical microleakage between groups A1 and B1, A2 and B2, C1 and C2 (P > 0.05). The apical microleakage in the group A1 was significantly higher than that in the group A2, and the group B1 also showed higher apical microleakage than the group B2 (P < 0.05). Our findings suggest that either immediate or delayed post space preparation exposes little influence on the apical microleakage after root canal filling with gutta-percha/mineral trioxide aggregate, which exhibits better apical sealing ability than the AH-plus.

BACKGROUND: Three-dimensional printing to prepare a digital model can improve the accuracy of orbital fracture repair, and has a good clinical prospect.OBJECTIVE: To evaluate the effect of individualized titanium mesh based on the digital model in the reconstruction of orbital fracture.METHODS: Twelve cases of orbital fracture were admitted at the Hospital of Stomatology, Southwest Medical University from January 2014 to October 2015. CT scanning in axial, coronal, sagittal planes and three-dimensional reconstruction were performed routinely in all cases preoperatively. Digital model was designed by 3D printing technology according to the CT data. Individualized titanium mesh was shaped based on the digital model and used to repair orbital fracture. The accuracy of the reconstructed orbit was assessed based on the postoperative CT scan.RESULTS AND CONCLUSION: Postoperative CT scans showed that the implanted individualized titanium meshes were capable of accurately reconstructing the fractured orbit in all the 12 patients, and there was no infection, and titanium mesh loosening, prolapse, and rejection. With healthy eyes as controls, 11 cases of eyeball retraction were corrected completely, and only 1 case was still under correction. Diplopia symptoms disappeared in the 8 of 9 cases, and relieved in the 1 of 9 cases. Seven cases of eye movement limitation recovered postoperatively. To conclude, the individualized titanium mesh has great accuracy to repair orbital fractures in patients without serious complications,which has achieved good clinical outcomes in the orbital reconstruction.

BACKGROUND:Interproximal contact loss is a complication of posterior crown/bridge restorations with a higher incidence, and it is induced by a variety of factors, such as age, tooth position, occlusal contact, tooth loss time, whether there is a repair on the free end.OBJECTIVE:To study and analyze the loss of interproximal contact between posterior crown/bridge restorations and adjacent teeth.METHODS: This was a single-center, prospective, observational clinical trial which has been completed at the Stomatology Hospital Affiliated to Southwest Medical University, China. Totaly 82 patients who had underwent posterior crown/bridge restorations from June 2015 to June 2016 in the Stomatology Hospital Affiliated to Southwest Medical University were selected. The time of denture wearing was 1-40 months, and the interproximal contact loss was defined by a 30 μm adjacent contact check or a flossing that can pass through the detection area with no resistance. Single-factor analysis and multiple correlation analysis were performed to analyze the interproximal contact loss at 1, 3, 6 months after posterior crown/bridge restorations. The study protocol was approved by the Ethics Committee of the Stomatology Hospital Affiliated to Southwest Medical University of China with an approval number of 2016024. All protocols were in accordance with Declaration of Helsinki, formulated by the World Medical Association. All patients and their relatives were informed of study protocols and provided a written informed consent prior to the beginning of the trial.RESULTS AND CONCLUSION:Of the 82 patients enroled, there were 41 adjacent contact zones with the emergency of interproximal contact loss, including 20 maxillary and 21 mandibular. It indicated that the interproximal contact loss had no association with the upper and lower jaw. Single-factor analysis showed that the time of denture wearing, tooth loss time and repair of the free end were confirmed as independent influencing factors for the complications (P < 0.05). Multiple correlation analysis found that free end repair and short-term tooth loss were independent factors of the interproximal contact loss. To conclude, free end repair and tooth loss time should be considered in the posterior crown/bridge restorations in order to minimize the occurrence of interproximal contact loss.

Objective To evaluate the role of caspase-1 in the spinal cord in a rat model of incisional pain.Methods Eighteen adult male Sprague-Dawley rats,weighing 250-280 g,in which intrathecal catheters were successfully implanted,were divided into 3 groups (n =6 each) using a random number table:incision pain group (group Ⅰ),incision pain plus dimethyl sulfoxide group (group ID) and incision pain plus caspase-1 inhibitor (Ac-YVAD-CMK) group (group IA).At 10 min before establishment of the model,0.9％ normal saline 20 μl was intrathecally injected in group Ⅰ,dimethyl sulfoxide 20 μl was intrathecally injected and then the catheter was washed with 0.9％ normal saline 10 μl in group ID,and Ac-YVAD-CMK 1 nmol/μl (dissolved in 20 μl dimethyl sulfoxide) and then the catheter was washed with 0.9％ normal saline 10 μl in group IA.The mechanical paw withdrawal threshold (MWT) of the ipsilateral hind paw was measured at 2 h before intrathecal catheterization (T0),3 days after intrathecal catheterization (T1) and 2,6,24 and 48 h after establishment of model (T2-5).The rats were sacrificed after the last measurement of pain threshold at T5,and lumbar enlargement segments of the spinal cord were removed for detection of caspase-1 (p20) expression and interleukin-1beta (IL-1β) content by Western blot and enzyne-linked immunosorbent assay,respectively.Results Compared with the baseline at T0,the MWT was significantly decreased at T2-5 in Ⅰ and ID groups (P＜0.05),and no significant change was found in MWT at T1-5 in group IA (P＞0.05).Compared with Ⅰ and ID groups,the MWT at T2-5 was significantly increased at T2-5,and the caspase-1 (p20) protein expression and IL-1β content were decreased in group IA (P＜0.05).There was no significant difference in MWT,expression of caspase-1 (p20) protein or IL-1β content between group Ⅰ and group ID (P＞0.05).Conclusion The activation of caspase-1 in the spinal cord can promote the release of IL-1β and thus is involved in the incision pain in rats.