Objective:To investigate the differences in gene expression levels in knee chondrocytes and chondrons in vitro.Methods:The chondrocytes and chondrons were isolated from full thickness of the 8-months ( n=5) rabbit knees cartilage. Chondrons from right knee were enzymatically isolated using 0.3% dispase and 0.2% collagenase-2 with shaking for 3 hours. Chondrocytes were isolated by 0.4% Pronase and 0.025% collagenase-2 from left knee. The mRNA levels in chondrocytes and chondrons were analyzed by quantitative real-time PCR, including matrix proteins [aggrecan(Agg), collagen(Col-2), Col-6A6, Col-10, Col-11], MMPs and inhibitors (MMP-1, MMP-3, MMP-9, MMP-13, TIMP-1, TIMP-2, TIMP-3), cytoskeletal proteins (Sox-9, vinculin, tubulin, actin), cytokines (IL-β, TNF-α). Analysis was performed using SPSS 16.0 statistical software, and the two-group comparisons were considered as significant by t-test at P<0.05. Results:Compared to the chondrocytes, the Agg [(5.78±0.90) vs (1.89±0.27), t=9.26, P<0.001], Col-2 [(6.29±0.76) vs (3.06±0.60), t=7.46, P<0.001], Col-6A6 [(0.89±0.18) vs (0.22±0.06), t=7.90, P<0.001], Col-10 [(3.83±0.76) vs (1.00±0.26), t=7.88, P<0.001] and TIMP-1 [(1.98±0.85) vs (1.03±0.34), t=2.32, P=0.049], TIMP-2[(3.46±1.50) vs (1.52±1.06), t=2.36, P=0.046], TIMP-3 [(3.96±0.50) vs (1.36±0.18), t=10.94, P<0.001], Sox-9 [(7.09±2.93) vs (3.24±0.77), t=2.84, P=0.022], vinculin [(3.42±1.69) vs (1.46±0.68), t=2.41, P=0.043], tubulin[(9.34±0.71) vs (2.35±0.80), t=14.61, P<0.001] showed higher expression in the chondrons. Compared to the chondrocytes, the MMP-1 [(1.02±0.30) vs (2.67±0.45), t=6.91, P<0.001], MMP-3 [(1.21±0.32) vs (2.52±0.79), t=3.44, P=0.009], MMP-13 [(1.23±0.34) vs (3.42±0.86), t=5.30, P=0.007], IL-1β [(1.02±0.14) vs (2.70±0.49), t=7.37, P<0.001], TNF-α [(0.99±0.08) vs (3.15±0.54), t=8.85, P<0.001] showed lower expression in the chondrons. There were no difference between chondrons and chondrocytes for Col-11, MMP-9, actin ( P>0.05). Conclusion:The gene expression of extracellular matrix components are higher and the gene expression levels of inflammatory factors and MMPs are decreased in chondrons compared with the chondrocytes, suggesting the chondrons have more multiplication potential as seeding cells for tissue-engineered cartilage.

Objective:To explore the short-term efficacy of totally laparoscopy pancreatoduodenectomy (TLPD) and open pancreatoduodenectomy (OPD) in the treatment of periampullary carcinoma.Methods:The clinical data of 50 patients with periampullary carcinoma in the First Hospital of Shanxi Medical University from June 2016 to March 2019 were retrospectively analyzed. According to the different surgical methods, the patients were divided into TLPD group (22 cases) and OPD group (28 cases). The perioperative and postoperative related indicators between the two groups were compared.Results:Both groups had successfully received the operation. The operating time in TLPD group was longer than that in OPD group, and the difference between the two groups was statistically significant [(665±213) min vs. (447±215) min, t = -0.356, P = 0.001]. The amount of intraoperative bleeding in TLPD group was less than that in OPD group, and the difference between the two groups was statistically significant [100 ml (50-325 ml) vs. 300 ml (100-500 ml), Z = -2.230, P = 0.026]. There were no significant differences in the proportion of intraoperative blood transfusion, lymph node dissection number, resected tumor diameter, postoperative diet restriction time, postoperative extubation time, postoperative hospital stay and the incidence of postoperative complication between TLPD group and OPD group (all P > 0.05). Conclusions:TLPD and OPD has a similar short-term efficacy in the treatment of periampullary carcinoma. The operating time of TLPD is longer than that of OPD, but TLPD can effectively control the intraoperative bleeding.

Objective:To investigate the short-term effects of articular injection of hyaluronic acid combined with glucocorticoid in patients with knee osteoarthritis.Methods:From October 2017 to June 2018, a total of 188 patients diagnosed with knee osteoarthritis received parallel articular injection. There were 60 cases with mild knee osteoarthritis, 72 with moderate and 56 with severe according to the WOMAC knee functional score. There patients were divided into group rank Ⅰ48 cases, Ⅱ 49 cases, Ⅲ 45 cases, Ⅳ 46 cases according to the knee joint X-ray Kellgren-Lawrence classification. The unified treatment regimen was 2.5 ml Sodium Hyaluronate (SHA) injection for the first time, SHA 2.5 ml and compound betamethasone injection (CBI) 1 ml for the second week, and 2.5 ml of SHA for the third week. WOMAC score and Lequesne index were used to evaluate joint function before the first injection and after SHA and SHA+CBI injection. The improvement rate of Lequesne index ≥30% or improvement rate of WOMAC score ≥25% was regarded as effective treatment.Results:Lequesne index and WOMAC score decreased gradually in the mild, moderate and severe groups after 3 weeks of injection. Among these patients, the improvement rates of Lequesne index after SHA injection and SHA+CBI injection were 36.44%±8.46% and 49.26%±13.75% in the mild group, 23.09%±12.61% and 30.66%±14.95% in the moderate group, and 10.50%±8.78% and 11.07%±6.52% in the severe group. The improvement rate of WOMAC score in the mild group after SHA injection and after SHA+CBI injection was greater than 25%. After SHA injection, the improvement rate of WOMAC score was 13.06%±10.21% in the moderate group, and 27.49%±13.61% after SHA+CBI injection. Those in severe group were all less than 25%. Kendall's staub correlation analysis results showed that there was a strong positive correlation between WOMAC function score and X-ray Kellgren-Lawrence classification ( r=0.744, P<0.001). The Lequesne index and WOMAC scores of the Kellgren-Lawrence X-ray classification decreased gradually after 3 weeks of injection. The improvement rate of Lequesne index period in group rank Ⅰ after SHA and SHA+CBI injection was 36.64%±10.05% and 52.00%±8.19%, respectively. That for group rank Ⅱ was 32.05%±8.09% and 41.95%±10.53%, group rank Ⅲ 16.93%±10.34% and 27.77%±10.25%, group rank Ⅳ 7.52%±5.53% and 7.60%±6.66%. The improvement rate of WOMAC score period in group rank Ⅰ after SHA and SHA+CBI injection was 29.48%±11.77% and 42.59%±13.55%, respectively. That for group rank Ⅱ was 26.72%±10.21% and 30.49%±16.90%, group rank Ⅲ 13.78%±5.96% and 23.05%±9.52%, group rank Ⅳ 4.77%±3.80% and 4.27%±4.23%. Conclusion:For mild or X-ray classification Ⅰ, Ⅱ knee osteoarthritis patients, articular injection SHA or SHA+CBI are effective. Further, SHA+CBI is better than single injection of SHA. SHA+CBI injection was effective for moderate knee osteoarthritis patients. For severe or X-ray classification Ⅲ, Ⅳ patients, SHA or SHA+CBI injection at interval are invalid.

Objective: To compare the short-term efficacy of robotic distal pancreatectomy (RDP) and laparoscopic distal pancreatectomy (LDP) in treatment of pancreatic body and tail cancer, and to explore the feasibility of RDP. Methods: The clinical data of 11 patients who received RDP and 26 patients who received LDP from January 2014 to May 2018 in the First Hospital of Shanxi Medical University were retrospectively analyzed. The operation indexes and the hospitalized cost of both groups were compared. Results: There were no significant differences in spleen-preserving rate, postoperative hospital stay, postoperative pain, intraoperative or postoperative blood transfusion, postoperative bleeding and pancreatic fistula between the two groups (all P > 0.05), but the amount of intraoperative bleeding in RDP group was less than that in LDP group, and the difference was statistically significant [(144±51) vs. (199±65) ml, t = -2.530, P = 0.016]. Compared with LDP group, the total hospitalization cost and operation cost of RDP group was increased [(75 000±14 000) yuan vs. (107 000±12 000) yuan; (21 000±9 000) yuan vs. (39 000±16 000) yuan; both P < 0.01]. Conclusion Both RDP and LDP are safe and feasible. LDP has the advantages of relative low cost and wide range of operations. RDP has obvious advantages in controlling intraoperative bleeding, but the high cost limits its further clinical promotion.

Objective To analyze the clinical feasibility and effectiveness of the "G"-shaped surgical approach in robotic pancreatoduodenectomy. Methods The clinical data of 17 patients who were undergoing robotic pancreatoduodenectomy at the First Hospital of Shanxi Medical University from June 2017 to March 2018 was analyzed. Results All the 17 robotic pancreatoduodenectomy operations via the "G"-shaped surgical approach were successful. The operationtime was (499 ±146) min (350-825 min), and the blood loss was (119±38) ml (20-500 ml). All surgical margins were negative. All patients recovered well after surgeries, and the postoperative hospital stay was (21 ±6) (14-36 days). However, one patient experienced secondary surgery due to bilioenteric anastomosis fistula, fortunately the surgical process went successfully. This patient had pancreatic leakage (class B) after surgery and was discharged with tubes after a conservative treatment, another patient had gastroplegia and recovered completely after conservative treatment. Conclusion It is a safe and feasible procedure to use the robotic pancreatoduodenectomy with the"G"-shaped surgical approach.

OBJECTIVE: To investigate the characteristics and regularity of ADR induced by antineoplastic drugs and provide reference for safe drug use in clinic. METHODS: ADR reports induced by antineoplastic drugs reported by 108 hospitals during Jan. 2009-Dec. 2016 were collected from PLA ADR Monitoring Center. ADR reports were analyzed respectively in respects of types of ADR reports, patients ' gender and age, administration route, occurrence time, types of antineoplastic drug, the situation of patients suffering tumor, systems/organs involved in ADR, clinical manifestations, outcome, etc. RESULTS: Among 15 183 ADR reports, there were 462 cases of new ADR and 2 873 cases of severe ADR; there were 8 039 male (52. 95％) and 7 144 female (47. 05％). The proportion of severe ADR in female (20. 00％) was significantly higher than male (17. 96％), with statistical significance (P=0. 001). ADR was mainly induced by intravenous administration (90. 53％), and mainly occurred 2-＜7 d after medication (23. 00％). Top 3 drug categories in the list of ADR were platinum antineoplastic drugs (25. 63％), plant-derived antineoplastic drugs and its derivative (24. 42％) and anti-metabolism drugs (18. 50％). Male patients mainly suffered from lung cancer, colorectal cancer and gastric cancer; female patients mainly suffered from breast cancer, lung cancer and colorectal cancer. Systems/organs involved in ADR were gastrointestinal system, hematological system and systemic damage. Main clinical manifestations were nausea, vomiting, myelosuppression, skin rash and fever. Totally 92. 57％ of ADR cases were cured and recovered after treatment, and 5 cases died. CONCLUSIONS: Antineoplastic drugs have high incidence of ADR with serious damage. Clinic should strengthen the monitoring of key population and key drugs so as to reduce the occurrence of ADR.

Objective To explore the histological properties of isolated chondrons and chondrocytes from rabbit knee cartilage,and to determine if these properties vary with age.Methods Three groups of rabbit knees were evaluated according to different age:(1) young (2 months,n=10);(2) adult (8 months,n=10);and (3) old (31 months,n=10).The cartilage structure,proteoglycan,collagen-2,and collagen-6 content were determined by light microscopic using hematoxylin-eosin (HE),Toluidine Blue,and col-2,6 staining.The chondrons were enzymatically isolated using 0.3 g/L dispase and 0.2 g/L collagenase-2 by shaking for 3 hours.The morphology and composition of isolated chondrons were observed by HE and collagen-6 immunostaining staining after overnight coverslip monolayer culture under a microscopy.Results The chondrocytes became sparser and the total content of proteoglycans and collagen-2 were decreased in the articular cartilage with age.Compared to the chondrocytes,the surrounding rim or capsule was more obvious in the isolated chondrons,and they exhibited obvious differences in shape.The cells within one cluster from different age groups were similar to the morphology observed in cartilage in situ.The adult and old chondrons generally possessed a thicker pericellular matrix with more enclosed cells,and the chondrons contained more cells can reach 47％.Conclusion These findings further suggest that the properties of the chondrons and pericellular matrix have an important influence on the biomechanical microenvironment of the knee joint cartilage degeneration that occurs with age.

Objective To explore the relationship among the expression level of MMP?13, AGG and Col?II in the chon?drocytes caused by nutritional deficiencies in rabbits. Methods 30 New Zealand white rabbits were randomly divided into autolo?gous chondrocyte transplantation group (control group, n=10), nutrition block group (surgery group, n=10), and peripheral nutrition block group (sham surgery group, n=10). 4 weeks after treatment, the rabbits were sacrificed for undergoing the observations on general and histological level;real?time PCR assay was employed for testing the expression level of MMP?13, AGG and Col?II;cel?lular apoptosis percentage was observed through TUNEL stain. The relationship among the apoptosis level, cartilage cells histologi?cal Mankin score as well as the expression level of MMP?13, AGG and Col?II were analyzed. Results Based on the Mankin score, there was a statistic difference between surgery group and control group. On the other side, there were no statistic differenc?es between sham surgery group and control group. 4 weeks after treatment, surgery group presented a higher apoptotic percentage compared with control group;this value between sham surgery group and control showed no significant differences. There was an increased mRNA expression level of MMP?13 and a decreased mRNA expression level of AGG and Col?II in surgery group com?pared with control group;no statistic differences of all these values was found between sham surgery group and control group. His?tological Mankin score and apoptotic percentage presented positive correlation (r=0.922, P<0.001), the regression equation:Y=-0.548+0.404X, R2=0.844 (F=157.735, P<0.001); the mRNA expression level of MMP?13 and apoptotic percentage presented positive correlation (r=0.942, P<0.001), the regression equation:Y=0.951+0.116X, R2=0.883 (F=219.054, P<0.001). There was a nega?tive correlation between the mRNA expression level of MMP?13 and the mRNA expression level of AGG as well as Col?II (r=-0.956,-0.945, P<0.001). Conclusion Damage of cartilage cells causes the up?regulation of the MMP?13 expression which could ex?acerbate the degeneration of cells. It could induce the down?regulation of AGG and Col?II mRNA expression, which will cause the extracellular matrix synthesis disorder.

BACKGROUNDOsteoporosis (OP) is a common bone disease, which adversely affects life quality. Effective treatments are necessary to combat both the loss and fracture of bone. Recent studies indicated that caffeic acid phenethyl ester (CAPE) is a natural chemical compound from honeybee propolis which is capable of attenuating osteoclastogenesis and bone resorption. Therefore, this study aimed to investigate the effect of CAPE on bone loss in OP mice using micro-computed tomography (CT) and histology.

METHODSEighteen mice were prepared and evenly divided into three groups. The six mice in the sham+PBS group did not undergo ovariectomy and were intraperitoneally injected with PBS during the curing period. Twelve mice were ovariectomized (OVX) to induce OP. Six of them in the OVX+CAPE group were intraperitoneally injected with 0.5 mg/kg CAPE twice per week for 4 weeks after ovariectomy. The other six OVX mice in OVX+PBS group were treated with PBS. All the mice were sacrificed 4 weeks after ovariectomy. The tibias were bilaterally excised for micro-CT scan and histological analysis. The Mann-Whitney U test was used to test the statistical differences among groups.

RESULTSBone loss occurred in OVX mice. Compared with the sham+PBS group, mice in the OVX+PBS group exhibited a significant decrease in bone mineral density (BMD, P < 0.05), bone volume fraction (BV/TV, P < 0.01), trabecular thickness (Tb.Th, P < 0.05), and trabecular number (Tb.N, P < 0.01), as well as a non-insignificant increase in the number of osteoclasts (N.Oc/B.Pm). With CAPE treatment, the microarchitecture of the tibial metaphyses was significantly improved with a reduction of osteoclast formation. Compared with the OVX+PBS group, BV/TV in the OVX+CAPE group was significantly increased by 33.9% (P < 0.05).

CONCLUSIONCAPE therapy results in the protection of bone loss induced by OVX.

Animals ; Bone Density ; drug effects ; Caffeic Acids ; pharmacology ; Female ; Metabolism ; drug effects ; Mice ; Mice, Inbred C57BL ; Ovariectomy ; Phenylethyl Alcohol ; analogs & derivatives ; pharmacology ; Propolis ; chemistry ; Tomography, X-Ray Computed

BACKGROUNDChondrocytes' phenotype and biosynthesis of matrix are dependent on having an intact cytoskeletal structure. Microfilaments, microtubules, and intermediate filaments are three important components of the cytoskeletal structure of chondrocytes. The aims of this study were to determine and compare the effects of the disruption of these three cytoskeletal elements on the apoptosis and matrix synthesis by rabbit knee chondrocytes in vitro.

METHODSChondrocytes were isolated from full-thickness knee cartilage of two-month-old rabbits using enzymatic methods (n = 24). The isolated cells were stabilized for three days and then exposed to low, medium, and high doses of chemical agents that disrupt the three principal cytoskeletal elements of interest: colchicine for microtubules, acrylamide for intermediate filaments, and cytochalasin D for actin microfilaments. A group of control cells were treated with carrier. Early apoptosis was assessed using the Annexin-FITC binding assay by flow cytometry on days 1 and 2 after exposure to the disrupting chemical agents. The components and distribution of the cytoskeleton within the cells were analyzed by laser scanning confocal microscopy (LSCM) with immunofluorescence staining on day 3. The mRNA levels of aggrecan (AGG) and type II collagen (Col-2) and their levels in culture medium were analyzed using real-time PCR and enzyme-linked immunosorbent serologic assay (ELISA) on days 3, 6, and 9.

RESULTSIn the initial drug-dose-response study, there was no significant difference in the vitality of cells treated with 0.1 µmol/L colchicine, 2.5 mmol/L acrylamide, and 10 µg/L cytochalasin D for two days when compared with the control group of cells. The concentrations of colchicine and acrylamide treatment selected above significantly decreased the number of viable cells over the nine-day culture and disrupted significantly more cell nuclei. Real-time PCR and ELISA results showed that the mRNA levels and medium concentrations of AGG and Col-2 were significantly decreased for cultures treated with colchicine and acrylamide when compared with untreated cells at three, six, and nine days, and this inhibition was correlated with higher matrix metalloprotease-13 expression in these cells. Cellular proliferation, monolayer morphology, and matrix metabolism were unaffected in cytochalasin D-treated cells when compared with control cells over the nine-day culture period.

CONCLUSIONSThe disruption of the microtubulin and intermediate filaments induced chondrocyte apoptosis, increased matrix metalloprotease expression, and decreased AGG and Col-2 expression in rabbit knee chondrocyte cultures. Our findings suggest that microtubulin and intermediate filaments play a critical role in the synthesis of cartilage matrix by rabbit knee chondrocytes.

Animals ; Cartilage, Articular ; cytology ; metabolism ; Chondrocytes ; cytology ; Collagen ; metabolism ; Cytoskeleton ; metabolism ; Knee Joint ; cytology ; metabolism ; Microscopy, Confocal ; Rabbits