Objective To estimate the effects of camptothecin (CPT) on the expression of hypoxia-inducible factor-1α (HIF-1α) in HaCaT cells under hypoxic conditions (2％ O2),and to explore the potential therapeutic mechanism of topical CPT for psoriasis.Methods Some HaCaT cells were classified into 6 groups:5 test groups cultured in Dulbecco's modified Eagle's medium (DMEM) with the presence of CPT at 12.5,25,50,100 and 200 nmol/L respectively,and 1 control group cultured in DMEM with the presence of dimethyl sulfoxide (DMSO).All the 6 groups of cells were cultured under normoxic conditions for 12,24,48 or 72 hours or under hypoxic conditions for 12 hours.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferation of HaCaT cells after the normoxic culture,and Western blot to quantify the protein expression of HIF-1α after the hypoxic culture.Some HaCaT cells were classified into a normoxia group (21％ O2) and a hypoxia group (2％ O2),and each group was divided into a CPT (100 nmol/L)-treated subgroup and a non-intervention subgroup (treated with the vehicle).After 12-hour culture,real-time fluorescencebased quantitative PCR was performed to measure the mRNA expression of HIF-1α.Statistical analysis was carried out by using Levene'.s test,one-way analysis of variance,Dunnett-t test and factorial analysis with the SPSS16.0 software.Results After treatment with CPT at 12.5-200 nmol/L for 12-72 hours,the proliferation of HaCaT cells was inhibited in a concentration-and time-dependent manner.More concretely,the cell proliferation rates were inhibited by 17.66％ ± 6.46％,33.11％ ± 4.63％ and 56.31％ ± 1.69％ respectively in HaCaT cells after 12-hour treatment with 200 nmol/L CPT as well as 24-hour treatment with 100 and 200 nmol/L CPT compared with the control group at the corresponding time points (all P ＜ 0.05).The protein expression level of HIF-1 α was significantly decreased in HaCaT cells after 12-hour treatment with CPT at 12.5,25,50,100 and 200 nmol/L under hypoxic conditions compared with the control group (0.348 ± 0.065,0.261 ± 0.112,0.115 ± 0.043,0.045 ± 0.024 vs.1.445 ± 0.329,all P＜ 0.05).The mRNA expression level of HIF-1α (expressed as △Ct) in the CPT-treated subgroup and non-intervention subgroup was-5.575 ± 0.29 and -5.451 ± 0.21 respectively in the normoxia group,significantly higher than that in the hypoxia group (-6.543 ± 0.57 and -6.203 ± 0.31 respectively,F =29.856,P ＜ 0.05),while there was no significant difference between the CPT-treated and non-intervention subgroups (F =1.667,P ＞ 0.05).Conclusions CPT at 100 nmol/L could inhibit the expression of HIF-1α protein,but had no obvious effect on that of HIF-1α mRNA.

Objective To analyze the clinical characteristics of glenoid labral tear combined with rotator cuff tear so as to provide references for arthroscopic surgery.Methods A total of 50 patients (28 males and 22 females)at mean age of 45 years old(ranged from 30 to 58 years)were enrolled in the study.Clinical data including the mechanism and time of injury,symptoms,results of physical examination,X-ray and MRI were collected.Location and degree of rotator cuff and glenoid labral tears found during arthroscopic surgery were also recorded.There were 37 patients with anterior labral tears,eight with superior labrum tears and five with posterior labral tears.The rotator cuff tear was slight in 36 patients,moderate in 10 and severe in three.One patient had extra-large tear.Anterior part of rotator cuff was found torn in 21 patients,middle part in 16 and posterior part in 13.MRI of glenoid labral tears manifested disappearance of triangular appearance and enlarged joint capsule.MRI manifestations of rotator cuff tear were high signal in torn part and continuity loss.Results All patients were followed up for average 36 months(6-72 months),which showed satisfactory results.UCLA score was(15±3)points preoperatively and(32±3)points postoperatively(P＜0.01).Conclusions More attention should be paid to glenoid labral tear to avoid misdiagnosis and delayed treatment,for it is usually combined with rotator cuff tear.Location of rotator cuff tear has some relation to the range of glenoid labral tear,ie,the larger range of glenoid labral tear may induce more posterior part of rotator cuff torn.

ObjectiveTo investigate the feasibility of Legacy pedicle screw system plus minimally invasive technique in treatment of Tile type C pelvic fracture and evaluate its therapeutic effects.MethodsFrom January 2008 to March 2010, 12 patients with closed Tile type C pelvic fractures were treated with Legacy pedicle screw system plus minimally invasive technique.There were seven males and five females, at age range of 21-60 years (average 39.6 years).All patients had sacrum fractures of the posterior pelvic rings.The anterior pelvic ring fractures included unilateral ischiadic ramus fractures in four patients and bilateral ischiadic ramus fractures in eight.Three patients were combined with shock.The anterior pelvic rings were fixed with minimally invasive reconstruction plates and the posterior pelvic rings fixed with minimally invasive technique plus Legacy pedicle screw system.ResultsThe operation lasted for average 90 minutes (range, 80-110 minutes) , with average intraoperative blood loss of 105 ml (range, 80-150 ml).All patients were followed up for average 18 months (range, 6-32 months), which showed that all patients got satisfactory reductions and that the fractures were healed at one stage, with average period of 11.5 weeks (range, 9-13 weeks).There occurred no infections, failures of internal fixations or nerve injuries.According to improved Lindahl standard of pelvic injury function assessment, the result was excellent in 10 patients and good in two, with average score of 78.6 points.Conclusions Under master of the surgical indications, Legacy pedicle screw system plus minimally invasive technique takes advantages of minor trauma, less intraoperative fluoroscopy, short operation time, good curative resuhs and few complications in treatment of Tile type C pelvic fractures by fixation of the posterior pelvic rings and reconstruction of the anterior pelvic rings.

Objective To assess clinical outcomes of FasT-Fix technique in repair of meniscal injury under arthroscope. Methods A total of 36 cases (37 knees) of mensical tears were repaired with FasF-Fix technique. There were 26 males and 10 females, at mean age of 26 years (14-51 years). Eighteen meniscal tears were located in zone Ⅱ, 16 in zone Ⅲ and 3 in zone Ⅳ. The average length of the tear was 2.2 cm (1.0-3.0 cm). Results There were no any signs of early complications after sur gery. All cases were followed up For 6-26 months (mean 16 months). According to Lysholm scoring scale system ,the average score of operated knees was increased from preoperative 44.13+12.56 to postopera tive 80.24+12.67 (P<0.01). After operation, all the patients could move the knee joint at normal range, except for one case who had a limitation of 20 degree flexion, with no pain or interlocking symptom. All cases returned to original work and/or could continue sports game. Conclusion The FasT-Fix technique is a simple, safe and effective method for repair of properly selected meniscal tears.

Objective:To detect the change of composition of immune cells in the spleen of miR-126 knockdown(miR-126KD) mice and preliminarily explore its significance .Methods: The expression level of miR-126 in spleens of miR-126KD mice was deter-mined by Realtime PCR.And the total number of splenocytes was calculated.The pathologic morphology change of the spleen was observed by HE staining.And the changes on proportion of DCs ,macrophages cells ,γδ T cells and NK T cells,CD3+T cells and its subgroup ,as well as CD19+B cells in spleens of miR-126KD mice were analyzed by Flow cytometry and calculated respectively.The level of phosphorylated AKt and NF-κB was analyzed by Western blot assay.Results:Compared with those of WT mice ,the expression level of miR-126 decreased obviously ( P<0.05 ) and the total number of cells increased obviously in spleen of miR-126 KD mice ( P<0.05).Moreover,the pathologic morphology of miR-126KD mice was significantly changed.The proportion and number of NK T cells in the inherent cells were significantly increased ( P<0.05 ) , but the proportion of Mφcells were significantly decreased ( P<0.05 ) . Meanwhile,the proportion and number of CD3+T cells and CD4+T cells in the adaptive immune cells were significantly increased (P<0.05),while the total number of CD19+B cells were significantly decreased (P<0.05).Last,the level of phosphorylation Akt and NF-κB increased obviously in spleen of miRNA-126 knockdown mice.Conclusion: Change obviously on the composition of immune cells subsets in the spleen of miRNA-126 knockdown mice ,which it may be related to the altered level of phosphorylated AKt and NF-κB and provides a preliminary experimental basis for the further exploring the roles of miR -126 in immune response.

Objective To evaluate effects of camptothecin on the autophagy of HaCaT cells.Methods Some cultured HaCaT cells were divided into several groups to be treated with camptothecin at concentrations of 5,10,25,50,100 and 200 nmol/L,and 0.1％ dimethyl sulfoxide (DMSO) (control group),respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferative activity of HaCaT cells after 24-and 48-hour treatment,flow cytometry to evaluate cell apoptosis after 24-hour treatment,and Western blot analysis to measure the expression of autophagy-related proteins microtubuleassociated protein 1 light chain 3 (LC3) and p62.Some HaCaT cells were divided into 2 groups to be treated with 10 nmol/L camptothecin and 0.1％ DMSO for 24 hours,respectively.Then,indirect immunofluorescence assay (IFA) was performed to determine the LC3 expression.Results Camptothecin at low concentrations of 5 and 10 nmol/L had no significant effects on the proliferation and apoptosis of HaCaT cells.Compared with the control group,the cellular proliferative rates were significantly inhibited by (31.23 ± 1.00)％,(54.21 ± 8.10)％ and (66.75 ± 10.70)％ in the 50-,100-and 200-nmol/L camptothecin groups after 24-hour treatment respectively,and by (25.81 ± 5.99)％,(44.35 ± 5.32)％,(65.81 ± 8.28)％ and (73.23 ± 9.59)％ in the 25-,50-,100-and 200-nmol/L camptothecin groups after 48-hour treatment respectively (all P ＜ 0.001).After 24-hour treatment,the apoptosis rates were significantly higher in the 50-,100-and 200-nnol/L camptothecin groups (14.46％ ± 2.38％,19.15％ ± 1.59％,29.88％ ± 1.37％,respectively) than in the control group (3.80％ ± 0.13％,all P ＜ 0.001).After 24-hour treatment with 5 and 10 nmol/L camptothecin,the protein expression of LC3 Ⅱ was significantly up-regulated,while p62 protein expression was significantly down-regulated:IFA showed that the percentage of autophagosome-positive cells was significantly higher in the 10-nmol/L camptothecin group than in the control group after 24-hour treatment (36.67％ ± 4.55％ vs.6.23％ ± 0.92％,t =6.546,P =0.003).Conclusions Camptothecin at low concentrations of 5 and 10 nmol/L can induce autophagy of HaCaT cells,but has no obvious effects on cell proliferation and apoptosis.Camptothecin at concentrations of 50,100 and 200 nmol/L can inhibit cell proliferation,promote cell apoptosis,and decrease autophagy levels.

Objective To study the anti-tumor effects of alcohol extraction of Coix stalk objects on H22 tumor-bearing mice. Methods The animal model of tumor bearing mice with H22 ascitic tumor cells was established. Eighty-four model mice were randomly and equally divided into Coix stalk extract groups 1-5 (10, 8, 6, 4 and 2 g/kg), model control group and cyclophosphamide group. Mice were treated orally with Coix stalk alcohol extraction solution (10, 8, 6, 4 and 2 g/kg), cyclophosphamide 0.02 g/kg and normal saline once a day for 8 days for Coix stalk extract group, cyclophosphamide group and model control group. The mouse activity, the size and the appearance of time of abdominal swelling, and changes of hair, feeding and drinking water quantity were observed in groups of mice. The solid tumor mass was measured in H22 tumor-bearing mice. The tumor inhibitory rate, liver index, spleen index and thymus index were calculated. Results The axillary tumor muster was found first in model control group with the fastest growth, reduced independent activity, decreased appetite and dim in hair color, followed by the Coix stalk extract group 1 and group 2. The last was Coix stalk extract group 5 and cyclophosphamide group. The solid tumor mass were (0.47±0.18), (0.37± 0.13), (0.34±0.10), (0.30±0.11) and (0.28±0.09) mg for Coix stalk alcohol extract groups 1-5, which were significantly lower than those of model control group (0.60 mg±0.21 mg, F=5.700,P<0.05). The tumor inhibition rates were 21.67%, 38.33%, 43.33%, 50.00%, 53.33%and 60.00%in Coix stalk extract groups 1-5 and cyclophosphamide group. The liver index, spleen index and thymus index were lower in cyclophosphamide group and Coix stalk alcohol extract groups than those of model control group (except for the spleen index of Coix stalk extract group 1). The liver index was lower in Coix stalk ethanol extract groups than that of cyclophosphamide group. There were no significant differences in the spleen index, thymus index between Coix stalk ethanol extract groups and cyclophosphamide group. Conclusion Coix stalk alcohol extract has inhibitory effects on the tumor and liver damage in H22 mice.

[Objective] To investigate the effect of mitogen-activated protein kinases (MAPK) on cerebral ischemia induced by photothrombosis in Swedish amyloid precursor protein (APP/SWE) transgenic mice.[Methods] In APP/SWE transgenic mice and non-transgenic mice (n ＝ 12,respectively),photothrombotic stroke was induced,on 7 d after cerebral ischemia,the amount of the survival neuron in the penumbra was counted using Nissl staining (n ＝ 6),and the activities of p38MAPK and JNK were measured by Western blot (n ＝ 6).[Results] On 7 d after cerebral ischemia,ratio of amount of survival neuron over the penumbra in hippocampus in the ischemic side to that in the non-ischemic side in the non-transgenic mice group (78.3 ± 1.3)% was significantly higher than that in the APP/SWE transgenic mice group (70.5 ± 1.4)% (P ＜ 0.05);compared with the non-ischemic hemisphere,the activities of p38 MAPK and JNK increased significantly in the ischemic hemisphere in the APP/SWE transgenic mice group (P ＜ 0.05),whereas,there was no significant difference between ischemic and non-ischemic hemisphere in the non-transgenic mice group (P ＞ 0.05).[Conclusion] Photothrombosis causes more severe damage in the APP/SWE transgenic mice group than that in the non-transgenic mice group.The possible mechanism includes the increased activities of MAPK which enhance the process of neuronal cell apoptosis.

Objective To observe the clinical efficacy of moxibustion in treating perimenopausal syndrome (PMS).Method Totally 108 PMS patients of yang deficiency or yin deficiency constitution were randomized into a treatment group of 56 cases and a control group of 52 cases. The treatment group was intervened by moxibustion, while the control group was by medication. The modified Kupperman Index (KI), Hamilton Anxiety Scale (HAMA), Symptom Checklist-90 (SCL-90), and Self-rating Depression Scale (SDS) were observed before and after treatment for comparison.Result The KI score, HAMA score, SCL-90 total score, and SDS score were significantly changed in both groups after intervention (P<0.01,P<0.05). After treatment, the KI score, HAMA score, SCL-90 total score, and SDS score in the treatment group were significantly different from that in the control group (P<0.01,P<0.05).Conclusion Moxibustion is effective in treating PMS, and it can improve the anxiety and depression symptoms of the patients.

Objective To analyze the methods and effects for treating unstable posterior pelvic ring fracture combined with sacral nerve injury and further identify the relationships among the diagnostic methods,surgical approaches and clinical outcomes.Methods A total of 38 patients with posterior pelvic ring fracture combined with sacral plexus injury treated from January 2000 to January 2010 were enrolled in the study.There were 20 males and 18 females at an average age of 35 years (range,10 to 59 years).The causes of fractures included traffic injury in 20 patients,fall injury in 12,weighty object impingement injury in five,and stabbing injury in one.Classification of posterior pelvic ring fractures included fracture and dislocation of sacroiliac joints in eight patients,fracture of ilium wing in two and sacrum fracture in 28.According to the Denis typing of sacrum fractures,there was one patient with type Ⅰ fracture,14 with type Ⅱ fracture and 13 with type Ⅲ fracture.All 38 patients presented the decrease or loss of skin sensation around the lower extremities,perineal region and crissum.Simultaneously,30 patients suffered motor dysfunction of the lower extremities,while 20 patients had bladder and anus sphincter dysfunction or sexual disorder.Thirteen patients were suspected of sacral plexus avulsion and four of them were confirmed by myelography or MRI examination.All patients had at least one associated injury.The average ISS was 21.9 points ( range,9 to 47 points).Therapeutic methods were fracture reduction and fixation in the absence of nerve decompression for eight patients and nerve decompression for 30 patients including 26 patients being also managed by fracture reduction and fixation.Operation time ranged from 6 days to 6 months.The clinical outcomes were evaluated according to the British Medical Research Council (BMRC) evaluation criteria of sensation and movement function.Results Thirty-four patients were followed up for average 4.9 years ( range,1 to 10 years),during which their pelvis obtained stable recovery.The neurological outcome was excellent in two patients,good in four and unchanged in two in the nondecompression group and was excellent in 16 patients,good in nine and unchanged in one in the decompression group,with the decompression group superior to the non-decompression group ( P ＜ O.05 ).Conclusions For unstable posterior pelvic ring fracture combined with sacral nerve injury,nerve decompression and release combined with internal fixation can better improve the sacral nerve function and obtain good pelvic ring stability and is worth of clinical application.