Problem-based Learning(PBL)is a teaching approach focusing on students under the constructivism theory.In such a teaching approach,problems are essential.The thesis discusses the theoretical basis of the problem designment in PBL,namely the forms of problems,their functions,and principles in the problem designment under the constructivism theory.

Objective:To analyze and compare the clinical outcomes of total knee arthroplasty with patella arthroplasty and patella replacement. Methods: From May 1999 to January 2004, 251 knees in 203 patients (45 males and 158 females; age range: 25 to 91 years, average 62.8 years) with osteoarthritis (206 knees) and rheumatoid arthritis (45 knees) were evaluated using a questionnaire during physical examination. Totally 137 knees underwent patella arthroplasty and 114 underwent patella replacement, with a mean follow-up of 28.1 months. The patients were scored using KSS Score, Feller Score and British Orthopedics Association Patient-satisfaction Score. Results: Patella arthroplasty and replacement showed no obvious difference in post-operative improvement of the knee score, knee function score, patella function score and range of flexion and extent of knee. Compared with patella replacement group, patella arthroplasty group had a higher patient satisfaction rate (P

Objective To explore a new surgical procedure for phallic reconstruction. Methods Since Dec 2003 to Nov 2006,4 patients with penile loss were reconstructed by transferring a deep in-ferior epigastric perforator(DIEP)flap with implanting the 12th costal cartilage. Results All of the flaDs survived successfully.With the follow-up from 5 months to 3 years,the appearance oi the neope-nis and donor-site scar was satisfactory. No one developed fistula or urethral stenosls, All patients were reported the return of erogenous sensibility of the neopenis leading to orgasm and achmving Pene-tration during sexual intercourse. Conclusions This new procedure may provide an esthetlcaIlY ac-ceDtable and functional neophallus.It might be a new alternative in penile reconstruction.

0.05). Conclusion The defecation function was similar between CPP group and CJP group. Therefore, the coloplasty pouch seems to be superior because of feasibility, simplicity, and effectiveness.

Objective To investigate the effect of trichostatin A(TSA),a histone deacetylase inhibitor, on s-100-induced autoimmune hepatitis in mice.Methods A total of 26 six-week-old male C57BL/6 mice were randomly divided into control group,model group and TSA group(six in each group),and the rest 8 mice were used to extract the s-100 protein from liver tissue.Mice of model group and TSA group were injected intraperitoneally with s-100 with complete Freund's adjuvant to induce autoimmune hepatitis model.At day 21, TSA group mice were injected intraperitoneally with TSA 2 mg/(kg·d)for 7 days,and 0.9% sodium chloride solution containing 1% dimethyl sulfoxide was injected into the control and model group mice.Alanine transaminase(ALT)and aspartate aminotransferase(AST)in serum were measured and liver histopathology was observed.The protein levels of nuclear factor(NF)-κB and acetylated histone H3 in liver tissue were detected by Western Blot.The hepatic mRNA levels of NF-κB,HDAC3,toll-like receptor 4(TLR4)and TNF-α were measured by real-time PCR.ELISA was used to determine the TNF-α in serum.The results were analyzed with t test.Results The serum levels of ALT in control group,model group and TSA group were(122.00 ± 45.29),(459.33 ± 167.58)and(217.33 ± 49.25)U/L,respectively.The differences between model group and control group or TSA group were significant(t=4.76 and 3.41,respectively,both P<0.05).The serum levels of AST in control group,model group and TSA group were(127.83 ± 18.55),(389.67 ± 87.14)and (249.50 ± 71.72)U/L,respectively.The differences between model group and control group or TSA group were also significant(t= 7.20 and 3.04,respectively,both P< 0.05).The inflammation of the liver histopathology induced by s100 was alleviated by TSA.The relative expressions of NF-κB protein,NF-κB mRNA,TNF-α mRNA,HDAC3 mRNA and TLR4 mRNA in the liver tissue of model group mice were 2.43 ± 0.42,9.51 ± 0.36,10.53 ± 0.74,2.90 ± 0.22,and 4.50 ± 0.73,respectively,which were significantly higher than those of the control group(1.28 ± 0.49,1.28 ± 0.49,1.06 ± 0.14,1.72 ± 0.73,and 1.01 ± 0.31, respectively)(t=4.68,37.14,30.69,4.33 and 10.85,respectively,all P <0.05).In TSA group,the relative expressions of NF-κB protein,NF-κB mRNA,TNF-α mRNA,HDAC3 mRNA and TLR4 mRNA were decreased(1.30 ± 0.36,1.30 ± 0.36,2.38 ± 0.36,2.13 ± 0.32 and 2.40 ± 0.51,respectively),which were statistically lower than those in model group(t=4.58,30.62,24.12,2.81 and 5.81,respectively,all P<0.05).The serum TNF-α levels in control group,model group and TSA group were(122.37 ± 68.12), (1361.44 207.13)and(691.64 ± 162.12)ng/L,respectively.Compared with model group,the differences were statistically significant(t=13.92 and 6.24,respectively,both P<0.05).The relative expression of ac-H3 protein in the model group was 1.10 ± 0.08,which was higher than that in the control group 0.96 ± 0.17(t=2.27,P<0.05).That in TSA group was 1.30 ± 0.04,which was higher than the model group(t=-0.30, P <0.05).Conclusion Histone deacetylase inhibitor TSA alleviates autoimmune hepatitis by enhancing histone acetylation and inhibiting NF-κB and inflammatory factors.

Objective To evaluate therapeutic effects of bone marrow mesenchymal stem cells(MSC)derived exosomes on alcohol-induced liver injury.Methods Eighteen male C57BL/6 mice aged 6 to 8 week were randomly divided into control group,model group and exosomes group,with 6 mice in each group.The mice in the model group and the exosomes group were fed with Lieber-DeCarli ad libitum diet(Dyets Inc.)for 4 weeks,followed by gavage a bolus of ethanol at day 26,27 and 28.The mice in the control group matched the alcohol-derived calories with dextran-maltose.Meanwhile,the mice in exosomes group were injected with MSC-exosomes via the tail vein at day 14 and 26.After the experiment,serum levels of alanine aminotransferase(ALT)and aspartate aminotransaminase(AST)were detected,and the pathological changes of liver tissues were observed.The expressions of nuclear factor erythroid 2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),CD63,CD81,TSG101 and Cytochrome C were analyzed by Western blot,and mRNA levels of Nrf-2,HO-1,interleukin(IL)-10 and IL-17 were analyzed by real-time polymerase chain reaction(RT-PCR).The commercial kits were used to detect serum IL-10,IL-17 levels and liver tissue malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD)oxidative stress indicators.The numbers of regulatory T cell(Treg)and help T(Th)17 cells in the liver were analyzed by flow cytometry.One-way analysis of variance was used for comparison between groups.Results MSC-exosomes expressed positive markers CD63,CD81 and TSG101,but did not express the negative markers Cytochrome C.The serum ALT and AST levels in model group were(87.3±25.1)U/L and(223.2±43.5)U/L,respectively,while those in exosomes group were(47.7±12.0)U/L and(128.2±33.6)U/L,respectively.The differences between the two groups were both statistically significant(F=12.818 and 12.226,respectively,both P<0.05).Compared with control group,the SOD activity and GSH level in the model group significantly decreased with statistically significant differences(F=4.245 and 24.074,respectively,both P <0.05).Lieber-DeCarli ethanol feeding significantly increased intrahepatic MDA level in the model mice,which was reversed by MSC-exosomes supplementation,and the difference was statistically significant(F=36.675,P <0.05).Compared with control group,the intrahepatic protein expressions of Nrf-2 and HO-1 in model group were significantly decreased,while the expressions in exosomes group were obviously increased.The differences were statistically significant(F=33.623 and 14.960,respectively,both P <0.05).The expression trends of Nrf-2 and HO-1 mRNA were the same as those of protein expressions(F=20.784 and 276.336,respectively,both P <0.05).The proportions of liver Treg/Th17 in the control group,model group and exosomes group were 4.3±0.9,0.4±0.2,and 3.4±0.5,respectively.The differences among groups were statistically significant(F=64.227,P <0.05).Compared with control group,the serum protein and intrahepatic gene expression of IL-17 in the model group were significantly increased,which were reversed by MSC-exosomes treatment.The differences were statistically significant(F=15.581 and 40.095,respectively,both P<0.05).Serum IL-10 protein levels and intrahepatic IL-10 gene expression were significantly decreased after Lieber-DeCarli ethanol feeding,which were lower than the exosomes group.The differences were statistically significant(F=98.268 and 153.743,respectively,both P <0.05).Conclusions MSC-exosomes transplantation may relieve alcohol-induced liver injury.The mechanism could involve reduction of oxidative stress in the liver via regulating Nrf-2/HO-1 and normalizing the balance of Treg and Th17 cells.

Objective To evaluate the effect of bundles of care on the prevention of postoperative delirium among elderly patients with hip fractures.Methods A prospective randomized case control study was conducted on 80 patients (≥65 years old) with hip fractures from March 2017 to June 2017.The patients were divided into experimental group (n =43) and control group (n =37) according to the random number table method.The experimental group received bundles of care,while the control group received routine nursing.The patients in both groups were all surgically treated,and the confusion assessment method (CAM) was applied to diagnose delirium after surgery.Gender,age,fracture type,duration from injury to operation,internal fixation type,operation time,total amount of bleeding,visual analogue scale (VAS),incidence of delirium,complications,and adverse events were compared between the two groups.Results Among the 80 patients,there were 11 males and 69 females,aged averagely 79.3 years (range,65-95 years).No significant differences were found between experiment group and control group in terms of gender (male:6 cases vs.5 cases,females:37 cases vs.32 cases),age [(79.8 ± 7.8) years vs.(78.7 ± 8.9) years],cause of injury (traffic injuries:7 cases vs.4 cases;falling injuries:36 cases vs.33 cases),duration from injury to operation [(66.1 ±14.3)hours vs.(63.4 ±14.9) hours],fracture type (femoral neck:13 cases vs.10 cases;intertrochanteric:26 cases vs.24 cases;subtrochanteric:4 cases vs.3 cases),internal fixation type (artificial total hip:5 cases vs.5 cases;artificial femoral head:8 cases vs.5 cases;PFNA:29 cases vs.27 cases),operation time [(55.5 ± 16.8) minutes vs.(51.6 ± 17.0) minutes],total blood loss [(114.4 ± 73.9) ml vs.(108.1 ±72.0) ml] (P ＞ 0.05).After bundles of care intervention,the postoperative VAS [(2.2 ± 0.8) points vs.(4.3 ± 1.2) points],postoperative delirium incidence (9％ vs.32％),incidence of complications and adverse events (2％ vs.19％) in experimental group were significantly lower than those in control group (P ＜ 0.05).Conclusion Bundles of care can relieve the pain and effectively reduce the incidences of postoperative delirium,complications,and adverse events in elderly patients with hip fracture.

Objective To study the constituents of urinary stones in patients in Zhejiang,and analyze the composition difference between patients from northern Zhejiang province and southern Zhejiang province.Methods From October 2012 to October 2018,clinical data of 4 423 urinary stone patients treated in Sir Run Run Shaw Hospital,the Second Affiliated Hospital of Wenzhou Medical University,and Huzhou First People's Hospital was retrospectively analyzed.Infrared spectrum was used to analyze urinary calculi constituents.Among 4 423 patients,there were 3 041 males and 1 382 females,male to female ratio was 2.2∶ 1,and the mean age was (51.2 ±16.5) years.There were 2 974 northern Zhejiang patients and 1 449 southern Zhejiang patients.High incidence age group was 41-60 years [48.2％ (2 136/4 423)].The distribution characteristics of urinary calculi constituents in different groups of sex,age,and region were analyzed.Results Among the 4 423 cases,the mixed urinary stones were dominant in the urinary calculus [73.1％ (3 235/4 423)],in which,the most component was the calcium oxalate monohydrate + calcium oxalate dehydrate + carbonated apatite [36.2％ (1 604/4 423)];among the pure stones,the most component was the calcium oxalate monohydrate [16.3 ％ (719/4 423)].Carbonated apatite stones [70.1％ (970/1 382) vs.61.0％ (1 856/3 041),P ＜0.05] and magnesium ammonium phosphate stones [12.7％ (176/1 382) vs.4.9％ (150/3 041),P ＜ 0.05] were both more prevalent in females than males,but uric acid stones[10.6％ (325/3 041) vs.5.8％ (81/1 382),P ＜0.05] were more common in males than females.The proportions of calcium oxalate stones[90.6％ (961/1 060) vs.76.2％ (935/1 227),P ＜0.05],carbonated apatite stones [77.6％ (823/1 060) vs.50.7％ (623/1 227),P ＜ 0.05],and magnesium ammonium phosphate stones[9.1％ (97/1 060) vs.6.5％ (80/1 227),P ＜0.05] of 0-40 years group were all higher than ＞ 60 years group,however,uric acid stones were more frequent in ＞ 60 years group [3.5％ (38/1 060) vs.17.0％ (209/1 227),P ＜ 0.05].The proportion of calcium oxalate stones in southern Zhejiang was lower than northern Zhejiang [79.0％ (1 145/1 449) vs.89.4％ (2 661/ 2 974),P ＜ 0.05].However,carbonated apatite stones [71.5％ (1 037/1 449) vs.60.1％ (1 789/2 974),P ＜ 0.05],magnesium ammonium phosphate stones [15.1％ (220/1 449) vs.3.5％ (106/ 2 974),P ＜ 0.05],and uric acid stones [10.7％ (156/1 449) vs.8.4％ (250/2 974),P ＜ 0.05] were more prevalent in southern Zhejiang than northern Zhejiang.Conclusions The distribution of constituents of urinary stones in Zhejiang was different in genders,age,and regions.Carbonated apatite stones and magnesium ammonium phosphate stones were more prevalent in females and young people,and uric acid stones were more common in males and old people.Calcium oxalate stones were more common in youths.Moreover,calcium oxalate stones were more frequent in northern Zhejiang,and carbonated apatite stones,magnesium ammonium phosphate stones and uric acid stones were common in southern Zhejiang.

Objective To investigate the effect of high-fat diet(HFD)on liver damage caused by autoimmune hepatitis(AIH)in mice.Methods Fifty C57BL/6 male mice were divided randomly into four groups:standard chow(SC)group,HFD group,AIH + SC group and AIH+ HFD group.AIH model was built after feeding for one week and all mice were sacrificed after four weeks.Liver and spleen tissues and serum were collected. Liver histopathology was detected by HE staining. Serum alanine aminotransperase(ALT)and aspartate aminotransferase(AST)levels were measured.Western blot analysis and real-time polymerase chain reaction(PCR)analysis were used to test the expressions of NLR pyrin domain containing 3(NLRP3)and cysteinyl aspartate specific proteinase 1(Caspase-1).The concentrations of interleukin(IL)-6,IL-1β and tumor necrosis factor(TNF)-α were analyzed using enzyme linked immunosorbent assay technology.The amount of Th17 cells in spleen was analyzed by FACS.Means among groups were analyzed with one-way ANOVA.SNK-q analysis was used for groups with homogeneity of variance, while nonparametric test was used for groups with variance nonhomogeneity.Results Histologically,the H&E staining of liver tissue from HFD group showed adipose degeneration,and there was inflammation around vessel in AIH+SC group.Moreover,in AIH+HFD group,the inflammation was more serious with mildly adipose degeneration.Compared with SC group,serum levels of ALT and AST increased in HFD group and AIH +SC group,and greatest increase was observed in AIH+ HFD group.The differences were statistically significant(F=57.12 and 37.58, both P<0.05).The proportions of Th17 cells in SC group,HFD group,AIH+ SC group and AIH+HFD group were(2.98 ± 0.90)%,(6.89 ± 0.99)%,(6.47 ± 1.08)% and(9.96 ± 0.83)%, respectively.The differences among all groups were statistically significant(F=54.05,P<0.05).The concentrations of IL-1β,IL-6 and TNF-α in each group were as follows:SC:IL-1β[(7.62 ± 2.81)ng/L],IL-6 [(106.54 ± 53.08)ng/L],T NF-α[(107.26 ± 36.20)ng/L];HFD:IL-1β[(25.06 ± 7.09)ng/L],IL-6 [(220.11 ± 47.41)ng/L],TNF-α[(273.77 ± 33.62)ng/L];AIH+SC:IL-1β[(17.49 ± 5.68)ng/L],IL-6 [(260.73 ± 50.29)ng/L],TNF-α[(250.49 ± 81.63)ng/L];AIH+ HFD:IL-1β[(52.04 ± 10.22)ng/L], IL-6[(785.93 ± 70.91)ng/L],TNF-α[(913.97 ± 64.57)ng/L].The differences were statistically significant(F=44.66,242.15 and 233.49,respectively,all P<0.05).The expressions of NLRP3 and Caspase-1 were significantly increased in AIH+ HFD group than the other three groups(all P<0.05). Conclusions High-fat diet potentiates liver damage induced by autoimmune hepatitis,which might relate to the secretion of pro-inflammatory cytokines,the activation of Th17 cells and the NLRP3 inflammasome as well as pyroptosis.

Objective To study the immunoregulatory effect of sodium butyrate(NaB)on T helper cell 17(Th17)and the effect on toll-like receptor 4(TLR4)signal pathway in autoimmune hepatitis (AIH).Methods Fifty male C57BL/6 mice(6 weeks of age)according to the random number table method divided into control group(n=10),AIH group(n=10),NaB group(n=10)and high roughage diet(HRD)group(n=10),and the other ten mice were used to extract hepatic sytosolic S-100.After the establishment of AIH model,mice in NaB group were given sodium butyrate 500 mg/(kg·d)by gavage and those in HRD group were fed with high-fiber stuff.After 3 weeks of treatment,all the mice were sacrificed.The pathological change was observed.The serum levels of alanine aminotransferase(ALT), aspartate transaminase(AST),IL-17A and TNF-α,the proportion of Th17 in spleen,the expression levels of TLR4 and myeloid differentiation factor 88(MyD88)in liver were observed in each group.The tests of normality and homogeneity of variance were used to compare the means of each group.One-way analysis of variance and multiple comparative analyses were used in the statistical analysis.Results HE staining showed that inflammatory cell infiltration and hepatocyte necrosis were significantly reduced in mice treated with NaB and HRD compared to AIH group.Serum ALT levels in control group,AIH group,NaB group and HRD group were(24.833 ± 2.229),(88.333 ± 9.543),(27.167 ± 3.189)and (29.833 ± 6.113)U/L,respectively,while AST levels in each group were(97.00 ± 14.953),(285.000 ± 35.434),(139.500 ± 38.976)and(127.167 ± 28.687)U/L,respectively.The differences among groups were all statistically significant(F=156.49 and 44.118,respectively,both P<0.01).The proportion of Th17 in spleen and the expressions of the transcription factors retinoid-related orphan receptor gamma t in the spleen of the NaB group and HRD group were significantly lower than those of AIH group.The differences were statistically significantly(F=21.780 and 68.283,respectively,both P<0.05).The expressions of TLR4 and MyD88 in liver of AIH group were significantly higher than control group,but those were inhibited in NaB group and HRD group.The differences were statistically significantly(F= 26.235 and 28.293,respectively,both P<0.01).The expressions of IL-17 and TNF-α in liver and serum decreased in NaB group and HRD group.Conclusion NaB exerts an immunoregulatory effect in AIH and improves inflammatory reaction in liver.