ObjectiveThis study aims to examine the effect of Rhei Radix et Rhizoma-Coptidis Rhizoma on reducing insulin resistance in db/db mice by regulating the adenylate activated protein kinase （AMPK）/UNC-51-like kinase 1 （ULK1）/key molecule of autophagy， benzyl chloride 1 （Beclin1） pathway and elucidate the underlying mechanism. MethodSixty 6-week-old male db/db mice were studied. They were randomly divided into the model group， metformin group （0.26 g·kg^-1）， and low-， middle-， and high-dose groups （2.25， 4.5， 9 g·kg^-1） of Rhei Radix et Rhizoma-Coptidis Rhizoma. A blank group of db/m mice of the same age was set， with 12 mice in each group. After eight weeks of continuous intragastric administration， the blank group and model group received distilled water intragastrically once a day. The survival status of the mice was observed， and fasting blood glucose （FBG） was measured using a Roche blood glucose device. Fasting serum insulin （FINS） was measured using an enzyme-linked immunosorbent assay， and the insulin resistance index （HOMA-IR） was calculated. Hematoxylin-eosin （HE） staining was performed to observe the pathological changes in the liver of the mice. The protein expression levels of AMPK， Beclin1， autophagy associated protein 5 （Atg5）， and p62 in liver tissue were determined by using Western blot. The protein expression levels of autophagy associated protein 1 light chain 3B （LC3B） and ULK1 in liver tissue were determined using immunofluorescence. Real-time fluorescence quantitative PCR （Real-time PCR） was used to measure mRNA expression levels of AMPK， Beclin1， Atg5， ULK1， and p62. ResultCompared with the blank group， the model group exhibited a significant increase in body mass （P<0.01）. Additionally， the levels of FBG， FINS， and HOMA-IR significantly changed （P<0.01）. The structure of liver cells was disordered. The protein expression levels of AMPK， Beclin1， and Atg5 in liver tissue were significantly decreased （P<0.01）， while the expression level of p62 protein was significantly increased （P<0.01）. The expression levels of mRNA and proteins were consistent. Compared with the model group， the body mass of the metformin group and high and medium-dose groups of Rhei Radix et Rhizoma-Coptidis Rhizoma was significantly decreased （P<0.05）. FBG， FINS， and HOMA-IR were significantly decreased （P<0.05，P<0.01）. After treatment， the liver structure damage in each group was alleviated to varying degrees. The protein expressions of AMPK， Beclin1， Atg5， LC3B， and ULK1 were increased （P<0.05，P<0.01）， while the protein expression of p62 was decreased （P<0.01）. The expression levels of mRNA and proteins were generally consistent. ConclusionThe combination of Rhei Radix et Rhizoma-Coptidis Rhizoma can effectively improve liver insulin resistance， regulate the AMPK autophagy signaling pathway， alleviate insulin resistance in db/db mice， and effectively prevent the occurrence and development of type 2 diabetes.

Ameloblastoma is a benign tumor characterized by locally invasive phenotypes,leading to facial bone destruction and a high recurrence rate.However,the mechanisms governing tumor initiation and recurrence are poorly understood.Here,we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution.Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response(IR),bone remodeling(BR),tooth development(TD),epithelial development(ED),and cell cycle(CC)signatures.Of note,we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence,which was dominated by the EZH2-mediated program.Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids.These data described the tumor subpopulation and clarified the identity,function,and regulatory mechanism of CC ameloblastoma cells,providing a potential therapeutic target for ameloblastoma.

ObjectiveTo explore the mechanism of Dahuang Mudantang in alleviating the intestinal injury in the rat model of acute pancreatitis via the high-mobility group box 1 （HMGB1）/receptor for advanced glycation endproduct （RAGE）/nuclear factor-κB （NF-κB） signaling pathway. MethodOne hundred and twenty SPF-grade Wistar rats received retrograde injection of 5% sodium taurocholate into the biliopancreatic duct for the modeling of intestinal injury in acute pancreatitis. The rats were randomized into blank， model， low-， medium-， and high-dose （3.5， 7， 14 g·kg^-1， administrated by gavage） Dahuang Mudantang， and octreotide （1×10^-5 g·kg^-1， subcutaneous injection） groups （n=20）. The rats in blank and model groups received equal volume of distilled water by gavage. Drugs were administered 1 h before and every 12 h after modeling， and samples were collected 24 h after modeling. The general status of the rats was observed. The biochemical methods were employed to measure the levels of amylase （AMS） and C-reactive protein （CRP） in the serum. The enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6 in the colon tissue. The morphological changes of pancreatic and colon tissues were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to measure the expression levels of HMGB1， RAGE， inhibitor of NF-κB kinase （IKK）， and NF-κB suppressor protein α（IκBα）in the colon tissue. ResultThe rats in the model group showed poor general survival， writhing response， reduced frequency of defecation， and dry stool. The symptoms of rats in the model group were mitigated in each treatment group， and the high-dose Dahuang Mudantang showed the most significant effect. Compared with the normal group， the model group had elevated AMS and CRP levels （P<0.05）， which were lowered by Dahuang Mudantang （P<0.05）， especially that at the high dose （P<0.05）. Compared with the normal group， the modeling elevated that levels of TNF-α， IL-1β， and IL-6 （P<0.05）. Such elevations were lowered by Dahuang Mudantang （P<0.05）， and the high-dose group and the octreotide group showed better performance （P<0.05）. The modeling caused necrotic， congested， and destructed pancreatic and colonic tissues， which were ameliorated by the drugs， especially high-dose Dahuang Mudantang. Compared with the normal group， the modeling up-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）. Compared with the model group， Dahuang Mudantang and octreotide down-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）， and the high-dose Dahuang Mudantang demonstrated the best performance （P<0.05）. Western blot results showed a trend consistent with the results of Real-time PCR. ConclusionDahuang Mudantang can improved the general status， reduce inflammation， and alleviate histopathological changes in the pancreatic and colon tissues in the rat model of acute pancreatitis by inhibiting the HMGB1/RAGE/NF-κB signaling pathway.

OBJECTIVE To investigate the effects of Cistanches Herba polysaccharides （abbreviated as CDPS） on rats with constipation-predominant irritable bowel syndrome （IBS-C）. METHODS SD rats were divided into control group （20 rats） and modeling group. The modeling group was given 2 mL of normal saline at 0-4 ℃ intragastrically （once a day， for 14 consecutive days） to induce IBS-C model. After modeling， model rats were grouped into model group， positive control group （mosapride citrate， 1.35 mg/kg）， CDPS low-dose group （50 mg/kg） and CDPS high-dose group （100 mg/kg）， with 20 rats in each group. Administration groups were given corresponding drug solution intragastrically， and control group and model group were given a constant volume of water intragastrically， once a day， for 28 consecutive days. The fecal water content， serum content of 5-hydroxytryptamine （5-HT）， and charcoal powder propulsion rate of rats were determined in each group；the pathological morphology of colon tissue was observed， and mRNA and protein expressions of nerve growth factor （NGF） and tropomyosin receptor kinase A （TrkA） in colon tissue were detected. RESULTS Compared with control group， the fecal water content and carbon powder propulsion rate in the model group were decreased significantly （P＜0.05）； the rupture of mucosal muscle layer in colon tissue， significant infiltration of inflammatory cells and cellular edema were observed； the content of 5-HT in serum， and relative mRNA and protein expressions of NGF and TrkA were increased significantly （P＜0.05）. Compared with model group， the fecal water content and carbon powder propulsion rate of rats were increased significantly in administration groups （P＜0.05）， and pathological changes of colon tissue were relieved significantly， while the content of 5-HT in serum， the mRNA and protein expressions of NGF and TrkA in colon tissue were decreased significantly （P＜0.05）； among them， the above indicators inthe positive control group and CDPS high-dose group were generally close to those in the control group （P＞0.05）. CONCLUSIONS CDPS can alleviate the symptoms of IBS-C rats， which may be related to the inhibition of NGF/TrkA signaling pathway.

ObjectiveTo reveal the intervention effect of Dahuang Mudantang on pancreatic injury in rats with acute pancreatitis （AP） of dampness-heat in large intestine syndrome and explore its possible mechanism based on network pharmacology. MethodNinety-six SPF-grade Wistar rats were randomly divided into the following six groups： a blank group， a model group， low-， medium-， and high-dose Dahuang Mudantang groups （3.5， 7， and 14 g·kg^-1）， and a Qingyi Lidan granules group （3 g·kg^-1）， with 16 rats in each group. The AP model of dampness-heat in large intestine syndrome was induced in rats except for those in the blank group by "high-temperature and high-humidity environment + high-sugar and high-fat diet + retrograde injection of 5% sodium taurocholate into the pancreaticobiliary duct". The blank and model groups received equal volumes of distilled water by gavage， while the treatment groups were administered Dahuang Mudantang or Qingyi Lidan granules 1 hour before modeling， and 12 and 24 hours after modeling. Samples were collected 1 hour after the last administration. The general conditions of the rats were observed. The AP model of dampness-heat in large intestine syndrome was evaluated. Serum amylase （AMS） and C-reactive protein （CRP） levels were determined using biochemical methods. Pancreatic tissue morphology was observed using hematoxylin-eosin （HE） staining. Network pharmacology was employed to predict potential targets of Dahuang Mudantang in the intervention in AP， and molecular biology technique was used to verify relevant targets. ResultCompared with the blank group， the model group exhibited lethargy， unkempt fur， loose and foul-smelling stools， elevated anal temperature with arching and twisting reactions， significantly increased serum levels of AMS and CRP （P<0.05）， abnormal pancreatic ductules， disordered interlobular spaces， and inflammatory cell infiltration in histopathological examination， as well as pathological changes including pancreatic acinar cell swelling， congestion， and necrosis. Compared with the model group， the treatment groups showed varying degrees of improvement in general survival conditions， reduced twisting reactions， visibly improved stool characteristics， reduced pancreatic tissue edema and necrosis， decreased serum AMS and CRP levels （P<0.05）， with the high-dose Dahuang Mudantang group showing the most pronounced effects （P<0.05）. Network pharmacology prediction indicated that hederagenin， β-sitosterol， and quercetin were the most widely connected active compounds with disease targets. Protein-protein interaction （PPI） network analysis revealed that protein kinase B （Akt）， tumor protein P53 （TP53）， tumor necrosis factor （TNF）， interleukin-6 （IL-6）， transcription factor （JUN）， vascular endothelial growth factor α （VEGFα）， interleukin-1β （IL-1β）， and vascular cell adhesion molecule-1 （VCAM1） were key targets in the "drug-disease" interaction. KEGG enrichment analysis suggested that the response of the mitogen activated protein kinase （MAPK） signaling pathway might be a core mechanism for DHMDT in the intervention in AP. Molecular biology analysis showed that compared with the blank group， the model group had significantly increased levels of TNF-α， IL-6， and VCAM-1 in pancreatic tissue （P<0.05）， as well as significantly elevated expression levels of p38 mitogen-activated protein kinase （p38 MAPK）， mitogen-activated protein kinase-activated protein kinase 2 （MK2）， and human antigen R （HUR） genes and proteins （P<0.05）. Compared with the model group， the treatment groups exhibited decreased levels of TNF-α， IL-6， and VCAM-1 in pancreatic tissue （P<0.05）， reduced expression levels of p38 MAPK， MK2， and HUR genes and proteins， with the high-dose Dahuang Mudantang group showing the most pronounced effects （P<0.05）. ConclusionDahuang Mudantang activates and regulates the p38 MAPK/MK2/HUR signaling pathway to suppress the release of inflammatory factors， thereby improving pancreatic injury.

Objective:To compare the difference in clinical efficacy between a free wrist crease flap pedicled with superficial palmar branch of the radial artery flap (SPBRAF) and a traditional free toe flap (TFTF) in reconstruction of hand soft tissue defects, and to provide reference for the treatment of small-to medium-sized hand soft tissue defects.Methods:Data of 37 patients who received hand surgery in Department of Hand Surgery, No.971 Hospital of the PLA Navy from December 2016 to December 2019 for small-to medium-sized hand soft tissue defects were retrospectively studied. Among the 37 patients, there were 32 males and 5 females, aged between 18 and 65 years old, with 41.5 years old in average. According to the reconstructive surgical procedure, patients were divided into SPBRAF group (22 cases) and TFTF group (15 cases). Regular follow-ups were conducted after surgery. The difference in curative effect at the last follow-up between the 2 groups was evaluated by the comparison of data acquired in follow-up. SPSS 25.0 was used to analyse the data statistically. The evaluation indicators included flap survival, long-term recovery of flap, recovery effect at donor site, total active movement(TAM) of the affected digit, time of hospital stay and the time return to work. P<0.05 was considered a statistically significant. Results:All free flaps survived. All patients were entered 6-18 (mean, 10) months of postoperative follow-up to comprehensively evaluate the therapeutic effect. According to the Evaluation Trial Standards of Upper Limb Partial Function of Hand Surgery of Chinese Medical Association, in the SPBRAF group, 20 flaps were found in excellent, and 2 in good; in the TFTF group, 14 flaps were found in excellent, 1 in good. There was no statistical difference between the 2 groups( P>0.05). The colour, texture and thickness of flaps between the 2 groups were either in excellent or good. There was no statistical difference between the 2 groups( P>0.05). TPD in the TFTF group (5-6 mm) was better than that in SPBRAF group (6-7 mm) with statistical difference between the 2 groups ( P<0.05). Texture at donor sites between the 2 groups was either in excellent or good ( P>0.05). In terms of appearance, sensation and recovery time of donor site, it was found that the SPBRAF group(mean, 6 weeks) was significantly better than those in the TFTF group(mean, 8 weeks) and there was statistical difference between the 2 groups ( P<0.05). In terms of recovery of TAM in single-digit, excellent or good were shown in both groups and there was no statistical difference between the 2 groups ( P>0.05). In terms of hospitalisation and time for return to work, the SPBRAF group(mean, 8 days and 17 weeks) was significantly better than that of TFTF group(mean, 12 days and 24 weeks), and there was statistical difference between the 2 groups ( P<0.05). Conclusion:SPBRAF has an ideal effect on reconstruction of small-to medium-sized hand soft tissue defects in hand. Although the flap is still inferior in sensation and appearance compared with the TFTF, the advantages in terms of donor site recovery, patient satisfaction of the donor site and reduced time of hospitalisation and return to original work are more obvious. SPBRAF provides a good complement to surgical procedures reconstructing a digit defect.

Objective To investigate the anatomy characteristics of saphenous artery(SA) and its perforator vessels in mini pig,and to provide a new perforator flap animal model.Methods Between May,2018 and August,2018,6 7-months-old mini pigs weight 25 kilograms were sacrificed by blooding under general anesthesia.The bilateral external iliac arteries were injected with a mixture of latex-lead oxide mixture and underwent CT-scanning to provide 3-dimensional reconstruction.The origin,diameter,courses and distribution of the SA and perforators of both hind limbs were observed.Results One of the hind limbs was abandoned due to the leakage of perfusion fluid.The other 11 hind limbs were available.The femoral artery passed through the medial femoral muscle and sartorius muscle in the middle of thigh,and run between the medial femoral muscle and gracilis muscle above the knee.Below the knee,it run on the surface of the medial leg muscles.A number of small branches were distributed in the tarsal joint and calcaneus at the medial malleolus,and there were abundant communicating branches with the posterior tibial artery.The average length of SA was (14.86±0.76) mm.The outer diameters of SA at initiative,medial tibial condyle and medial malleolus were(1.73±0.15) mm,(1.50±0.12) mm and (1.30±0.13) mm,respectively.There were a total of 79 perforators which were identified,with a mean of (7.09±1.16) perforators per hind limbs.And the average outer diameter was 0.10-0.78 (0.40±0.13) mm.Conclusion The SA constantly exist with good outer diameter.The number of perforator is abundant.SA can be used as a reliable animal model for studying and training of perforator flap.

According to the course teaching requirements of psychiatry,a serial teaching cases of case-based learning (CBL) is built based on cases from psychology department to assist in the theory teaching of Psychiatry.During the teaching process,the scenes where patients came to the psychology department were simulated and the information about the cases was provided step by step.For each step,students would be asked to discuss a series of questions regarding the cases in different groups.Along with CBL,students could thoroughly learn the main points of diagnosis and principles of treatment for common psychiatric disorders.The teaching effect showed that students' involvement in CBL courses,the interests in learning,self-study ability,independent thinking ability,capacity for self-expression,ability of creative thinking and comprehensive analysis and problem solving ability.were greatly improved.Most students were interested in CBL and it produced fruitful teaching results.

OBJECTIVE To investigate the effect and related mechanism of all￣trans retinoic acid (atRA) exposure on osteogenic differentiation of mouse embryonic palate masenchymal cells MEPM. METHODS MEPM were cultured in osteogenic medium (OM) with atRA 0.1 and 1.0 μmol??L-1 for 1, 3,5, 7 and 9 d. MTT assay was performed to measure the cell viability. The alkaline phosphatase (ALP) activity was measured by chemical colorimetry. The cells were stained using the Von￣Kossa technique to detect the formation of mineralization nodules after 21 d of culture. RT￣PCR was performed to determine expression Runx2, osteopontin, bone morphogenetic protein receptor ( Bmpr) 1b, Bmpr2 and Smad5 mRNA. RESULTS The result of MTT on 9 d showed that, compared with normal control group, the cell viability of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups decreased significantly(P<0.01). Compared with normal control group, ALP activity of OM group increased significantly(P<0.05), while the ALP activity of OM+atRA 0.1 and 1.0 μmol??L-1 groups was lower than OM group(P<0.05). On 21 d, the Von￣Kossa stai￣ning results showed that the percentage of mineralization nodules formation of OM+atRA 1.0 μmol??L-1 group was (3.65±1.24)%, which was significantly lower than that of OM group(10.33±2.29)%(P<0. 05). On 9 d, the relative Run expression of OM group was the highest one in the four groups, while at￣RA 1.0 μmol??L-1 treatment negatively regulated 20% in comparsion with OM group(P<0.05). Compared with normal control group, the mRNA expression of osteopontin of OM, OM+atRA 0.1 and 1.0 μmol??L-1 groups increased significantly(P<0.05); BDNF mRNA expression of OM group was 2.6￣fold to normal control group, while that of OM+atRA 1.0 μmol??L-1 group was 33% to OM group(P<0.05) . The level of Smad5 mRNA of OM+atRA 1.0 μmol??L-1 group was significantly lower than that of OM group(P<0.05). CONCLUSION atRA Might inhibit osteogenic differentiation of MEPM by down￣regulated the expression of Bmpr1b.

Objective:To study the influence of all-trans retinoic acid (atRA)on craniomaxillofacial development of C57 mice. Methods:Pregnant C57BL mice were divided into 4 groups(n =5)at gestation day (GD)1 0.Mice in three atRA-induction groups were given atRA of 60,80 and 1 00 mg/kg,respectively.The mice in control group were given the equivalent volume of corn oil.All pregnant mice were sacrificed at GD1 9 and the embryos were collected.Stereo microscope was used to observe the craniomaxillofacial morphology.Standardized radiographs were taken and cephalometric analysis was performed.Results:The embryonic body length and body mass of control group surpassed those of 80 and 1 00 mg/kg atRA groups(P <0.05,P <0.01 ).atRA induced craniomaxillofacial malformations and maldevelopment.The mice induced by atRA exhibited a shorter mandibular body and more retrusive position of max-illary and mandibular(∠NAK and ∠NBD)when compared with their norm(P <0.01 ).Significant decrease in craniofacial length (Op-Rh)was observed in all atRA-induced groups(P <0.01 ).Decreases in cranial vault height(Fp-Os)and cranial vault length(Pa-Na)dimensions were observed in 80 and 1 00 mg/kg atRA groups(P <0.05,P <0.01 ).Conclusion:Exogenous atRA dose-depend-ently induces retardation of craniomaxillofacial morphology in embryo of C57BL mice by inhibition of the sagital and vertical dimension development of the bone.