Objective To isolate human dental pulp stem cells and cultivate them in 2D and 3D.Methods Dental pulps were separated from the second premolars and digested by collagenase type Ⅰ and dispase.Dental pulp stem cells were obtained and identified.The proliferation of dental pulp stem cells cultivated in 2D and 3D was compared.Results There were clonogenic cells in human dental pulp.Some dental pulp stem cells could be differentiated into myocytes and odontoblasts.The cell density was increased to 4.12 times of the primary density in 2D culture and 11.06 times in 3D culture.After 3D culture,the biological properties of dental pulp stem cells remained.Conclusion Human dental pulp stem cells were successfully isolated and cultivated from the second premolars.3D culture is a suitable way to expand dental pulp stem cells.

Objective To examine the effect of epigallocatechin gallate (EGCG) at different concentration (0,10,25, 50 and 100μmol/L ) on proliferation rate and apoptosis of nasopharyngeal carcinoma cell line C666-1 in vitro, and elucidate the role of silent information regulator 1 (SIRT1). Methods The proliferation rate in vitro of C666-1 cells stimulated by EGCG at increasing concentrations (0,10,25,50,and 100 μmol/L)for 24 h or at concentration of 50μmol/L for 0,6,12 h and 24 h were detected by cell counting kit-8 (CCK-8) assays. Cell were treated with EGCG at concentration of 0,25, 50 and 100 μmol/L for 24 h, cell apoptosis was anylysed by TUNEL assay and expression levels of SIRT1 protein was detected by western blotting. Results EGCG suppressed cell proliferation of C666-1 cell line in a concentration-dependent manner at concentration of 0 ,10,25,50,and 100μmol/L, and in a time-dependently when treated with 50 μmol/L for 12 to 24 h(P<0.05). After treated for 24 h with different concentration of EGCG at 0、25、50、100 μmol/L, cell apoptosis increased at concentration of 50 to 100μmol/L and expression of SIRT1 decreased in a concentration-dependently (P<0.05). Conclusion EGCG induced cell apoptosis of C666-1 cells by down-regulating SIRT1 expression.

Objective To observe ghrelin expression in gastric tissue and serum leptin level of the early over-fed rats given ω-3 polyunsaturated fatty acids (PUFAs) diets after weaning,and explore the effects of early over-feeding and diets intervention on the metabolic syndrome in adult rats.Methods Male Sprague-Dawley rats were divided into normal feeding group (NF group,10 per litter) or over-feeding group (OF group,3 per litter) in postnatal day 3,and then different diets were given after weaning (postnatal day 21).The OF group was separately given conventional diet (OF + Con group),high-fat diet (OF + HF group),or ω-3 PUFAs (OF + ω-3 group) ; while the NF group was separated into NF + Con group and NF + HF group.Body weight and food intake were recorded every week.In week 6 and week 16,glucose tolerance test was perforfmed.Serum leptin,ghrelin,and triglyceride were assayed by enzyme-linked immuno-absorbent assay.Ghrelin mRNA and protein levels in gastric tissue were quantified by real-time PCR and immunohistochemistry.Results At week 16,energy intake,body weight and glucose intolerance in OF + HF group were significantly higher than in NF + HF group (t =-3.453,P =0.014 ; t =-6.406,P =0.000 ; F =16.249,P =0.000),and serum triglycerides,ghrelin mRNA of gastric tissue were significantly higher than those of OF + Con group (t =4.72,P =0.005 ; t =8.486,P =0.000).At week 16,the serum leptin level in OF + Con group was higher than that in NF + Con group (t =-3.274,P =0.031),also higher in OF + HF group than that in OF + Con group (t =3.028,P =0.014).There were no significant differences in serum ghrelin and the area of ghrelin immuno-positive cells in the gastric tissue among groups.The above indicators in OF + ω-3 group were not different from those of NF + Con group.Conclusions Over-feeding during the lactation period may lead to high susceptibility to obesity and disordered glucose and lipid metabolism.It can also increase serum leptin and ghrelin mRNA expression in gastric tissue,aggravate leptin resistance and feeding control disorders.Dietary ω-3 PUFAs offered protection against excessive accumulation of adipose tissue,glucose intolerance,leptin resistance,and maintained normal levels of leptin and ghrelin.

Objective To evaluate the effects of dexmedetomidine on perioperative inflammatory response and cellular immune function in patients undergoing posterior lumbar interbody fusion. Methods Eighty American Society of Anesthesiologists physical statusⅠorⅡpatients of either sex, aged 40-60 yr, scheduled for elective posterior lumbar interbody fusion, were divided into dexmedetomidine group(group Dex)and control group(group C)using a random number table with 40 patients in each group. In group D, dexmedetomidine at a loading dose of 0.5 μg∕kg was intravenously infused starting from 10 min before anesthesia induction, followed by continuous infusion of 0.5 μg·kg-1·h-1until 15 min before the end of operation. The equal volume of normal saline was given at the same time points in group C. Before induc?tion, at 30 min after beginning of operation and at 1 h and 1, 3 and 5 days after the end of operation (T1?6), arterial blood samples were collected for determination of the plasma CD42a+∕CD14+ratio, HLA?DR+∕CD14+ratio, concentration of C?reactive protein(CRP)and white blood cell(WBC)count. Re?sults Compared with the baseline at T1, the plasma CD42a+∕CD14+ratio was significantly increased at T2?6, the HLA?DR+∕CD14+ratio was decreased at T3?6, the plasma CRP concentrations were increased at T4?6, and the WBC count was increased at T3?5in group C, and the plasma CD42a+∕CD14+ratio was signifi?cantly increased at T6, the HLA?DR+∕CD14+ratio was decreased at T3?5, and the plasma CRP concentra?tions were increased at T2?5in group D(P<0.05). Compared with group C, the plasma CD42a+∕CD14+ra?tio was significantly decreased at T2?4, the HLA?DR+∕CD14+ratio was increased at T4?5, and the plasma CRP concentrations and WBC count were decreased at T2?5in group D(P<0.05). Conclusion Dexme?detomidine can decrease perioperative inflammatory response and improve cellular immune function in the patients undergoing posterior lumbar interbody fusion.

Objective To explore the plasma levels of soluble CD40 (sCD40) and soluble CD40 ligand (sCD40L) in the patients with Alzheimer’s disease (AD) and those with amnestic mild cognitive impairment (aMCI). Methods The levels of plasma sCD40 and sCD40L were measured in 20 patients with AD, 35 patients with aMCI, and 32 cognitively normal controls (NC) using commercially available ELISAs. The cognitive function of AD and aMCI patients was mea?sured by mini-mental state examination (MMSE). Results There were significant differences in plasma sCD40 among AD, aMCI and NC groups (P<0.05) as the medians (the upper and lower quartiles) of plasma levels were 123.3 (97.4, 149.5) pg/mL, 102.9 (63.6, 124.0) pg/mL and 70.66 (51.0, 90.8) pg/mL, respectively. There were significant differences in plasma sCD40L among AD, aMCI and NC groups (P<0.05) as plasma levels were 537.0 (316.0, 1134.0) pg/mL, 316.0 (190.0,546.0) pg/mL and 167.0 (107.5,478.0) pg/mL. A negative correlation between the plasma concentrations of sCD40L and the MMSE scores was found in aMCI patients (r=-0.736, P<0.001). Conclusions There are relevant chang?es of plasma sCD40 and sCD40L levels in patients with AD and aMCI. The present results suggest that plasma levels of sCD40 and sCD40L may be appropriate biomarkers for AD patients and indicate that CD40-CD40L signaling may be in?volved in AD pathophysiology.

OBJECTIVE: To detect genomic copy number variations (CNVs) among 145 children with unexplained mental retardation/developmental delay (MR/DD) by using low-depth whole-genome copy number variation sequencing (CNV-seq). METHODS: Peripheral blood samples were collected from the patients and subjected to DNA extraction and CNV-seq. The results were analyzed by a combination of bioinformatic tools. RESULTS: Forty-nine patients were found to carry a total of 67 CNVs with an average size of 5.27 Mb. Among these, 22 patients were assessed to carry MR/DD-related CNVs involving 21 syndromes. This gave a diagnostic rate of 15.17%(22/145) for CNVs associated with unexplained MR/DD. The corresponding regions of the 22 MR/DD-related CNVs in the human genome covered 174 MR/DD-related pathogenic genes, which have mapped to 18 sections on 10 chromosomes. CONCLUSION Genomic CNVs-related microdeletions/duplications account for a significant proportion of unexplained MR/DD, for which CNV-seq can provide an accurate diagnosis.

Objective: To compare the effect of intranasal dexmedetomidine and oral chloral hydrate in deep sedation of children. Methods: The Pubmed, EMBase, CENTRAL (Issue 4, 2018), Web of science, CBM, Wanfang Data, CNKI and VIP databases from the inception to January 2019 were searched. Randomized controlled trials (RCTs) with dexmedetomidine and chloral hydrate as interventions were included and the data were analyzed by RevMam 5.3 and Stata 12.0 software. The success rate of deep sedation, the indicator of sedation onset time, the recovery time, the incidence of vomiting and bradycardia were compared. Results: A total of 7 RCTs involving 1 007 patients were included for analysis. The results showed that the success rate of deep sedation (OR=2.55, 95%CI:1.46-4.44, P<0.01) and the incidence of bradycardia (OR=4.42, 95%CI:1.82-10.74, P<0.01) in the dexmedetomidine nasal group were significantly higher than those in the chloral hydrate oral group. The recovery time was significantly shorter (MD=-16.41, 95%CI:-21.54-11.28, P<0.01) and the incidence rate of vomiting (OR=0.04, 95%CI:0.01-0.17, P<0.01) in dexmedetomidine nasal group was significantly lower than those in the chloral hydrate oral group. There was no significant difference in the indicator of sedation onset time (MD=-0.47, 95%CI:-2.71-1.22, P=0.46). Conclusion Compared with the traditional oral chloral hydrate, intranasal dexmedetomidine has a higher sedation success rate and shorter recovery time after sedation with a lower incidence of nausea and vomiting.

Objective:To establish a computer-aided design and 3D printing system for precise implantation of micro implant anchorage, and accurately calibrate the position and direction of micro implant implantation.Methods:A retrospective selection was conducted on 15 patients (30 in total) who underwent micro implant implantation surgery from the Department of Stomatology, the Affiliated Hospital of Jiangnan University from November 2019 to November 2021, including 6 males and 9 females, aged (17.1±6.3)years old. The preoperative patient was photographed with cone beam computed tomography (CBCT) and the collected DICOM data format was output. A 3D scan was performed on the patient′s preoperative analysis model to obtain the STL file of the model scan. The CBCT data and model data were fitted and matched using 3Shape Implant Studio software, and the thickness of the guide plate, the amount of undercut compensation, and the size of the key component collar were designed. The 3D printer was used for printing after resizing. Using the assist method to implant micro implants, CBCT was taken postoperatively to compare the preoperative design with the postoperative results.Results:After fitting the postoperative CBCT with the designed CBCT of the micro implant, it was found that the micro implant was consistent with the preoperative design, maintained a safe distance and parallel to the adjacent tooth root, and did not damage the maxillary sinus and other areas. No detachment of the micro implant anchorage was observed 1 or 3 months after surgery. The application of assisted micro implant anchorage 3D guide plate was reliable, with accurate implantation position and direction, and can be implanted in most parts of the oral cavity.Conclusions:The use of computer-aided design and 3D printing system to create an assistive micro implant anchorage 3D guide plate can accurately locate the position and direction of the micro implant, which is worthy of clinical promotion and application.

Objective:To juxtapose laparoscopic cholecystectomy combined with common bile duct exploration and stone extraction (LC+ LCBDE) against endoscopic retrograde cholangiopancreatography/sphincterotomy with laparoscopic cholecystectomy (LC+ ERCP/EST) in the therapeutic context of acute biliary pancreatitis.Methods:The clinical data of patients with acute biliary pancreatitis in Department of Hepatobiliary Surgery, Datong Third People's Hospital from January 2017 to January 2021 were retrospectively analyzed. A total of 44 patients were inrolled, including 23 males and 21 females, with the age of (60.6±11.7) years. Based on different treatment approaches, the patients were divided into the LC+ LCBDE group ( n=33) and the LC+ ERCP group ( n=11, LC+ ERCP/EST). Total bilirubin, direct bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood amylase, operation time, postoperative hospitalization stays, total hospitalization cost, postoperative anal exhaust time, and postoperative complications (bile leakage, fever, bleeding) were compared between the two groups. Results:There were no significant differences in preoperative total bilirubin, direct bilirubin, ALT, AST, and blood amylase between LC+ ERCP group and LC+ LCBDE groups (all P>0.05). In LC+ LCBDE group, operation time was 110.0 (96.3, 147.5) min, postoperative hospitalization time was 9.0 (7.5, 11.0) d, postoperative exhaust time was 2.0 (1.0, 2.0) d, and in LC+ LCBDE group, operation time was 60.0 (32.0, 65.0) min, postoperative hospitalization time was 7.0 (4.0, 8.0) d, postoperative exhaust time was 1.0 (1.0, 1.0) d. Comparisons with LC+ LCBDE group, LC+ ERCP group had shorter postoperative hospitalization stay and earlier postoperative exhaust time, the total hospitalization cost of LC+ LCBDE group was 23 829.3 (21 779.6, 27 221.9) yuan, which was higher than 36 894.8 (31 963.5, 41 172.2) yuan in LC+ ERCP group, and the differences were statistically significant (all P<0.05). Comparison of postoperative total bilirubin, direct bilirubin, ALT and AST between LC+ ERCP group and LC+ LCBDE group, with no significant difference(all P>0.05). No postoperative complications such as bile leakage, residual stones, fever and bleeding occurred in both groups. Conclusion:Compared with LC+ ERCP/EST, LC+ LCBDE in the treatment of acute biliary pancreatitis, although the operation time and hospital stay are longer, but the total hospitalization cost is less, there is no need for multiple operations, and it can be used as the first choice for acute biliary pancreatitis.

Objective:To evaluate the effect of esmolol on the expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) during cerebral ischemia-reperfusion (I/R) in rats.Methods:Forty-eight clean-grade healthy adult male Sprague-Dawley rats, were allocated into 3 groups ( n=16 each) using a random number table method: sham operation group (Sham group), cerebral I/R group (I/R group) and esmolol group (E group). Cerebral I/R was induced by 3 cycles of 20-min occlusion of bilateral common carotid arteries followed by 10-min reperfusion in anesthetized rats.Esmolol 200 g·kg -1·min -1 was intravenously infused for 1 h starting from 30 min before ischemia, and the model was established after 30-min infusion in E group.The equal volume of normal saline was given at 30 min before ischemia in I/R group.Bilateral common carotid arteries were only isolated but not clamped, and the equal volume of normal saline was given after isolating bilateral common carotid arteries in Sham group.Learning and memory function was tested by Morris water maze test before ischemia and at 1, 3 and 7 days of reperfusion.Rats were sacrificed after Morris water maze test, and the hippocampus was excised for determination of wet to dry weight ratio (W/D ratio), permeability of blood-brain barrier (using Evans blue method), expression of ERK1/2 mRNA (by real-time polymerase chain reaction ), and expression of p-ERK1/2 (by Western blot). Results:Compared with Sham group, the escape latency and swimming distance were significantly prolonged at 1, 3 and 7 days of reperfusion, the W/D ratio and EB content in brain tissues were increased, and the expression of ERK1/2 mRNA and p-ERK1/2 was up-regulated in I/R and E groups ( P<0.05). Compared with I/R group, the escape latency and swimming distance were significantly shortened at 1, 3 and 7 days of reperfusion, the W/D ratio and EB content in brain tissues were decreased, and the expression of ERK1/2 mRNA and p-ERK1/2 was down-regulated in E group ( P<0.05). Conclusion:The mechanism by which esmolol alleviates cerebral I/R injury and improves cognitive function is related to inhibiting the up-regulated expression of ERK1/2 in rats.