OBJECTIVE To explore the antitumor effect and mechanism of total alkaloids of Gelsemium elegans （TA） and sempervirine （SPV） in vitro. METHODS The effects of low， medium and high concentrations of TA （50， 100， 200 μg/mL） and SPV （10， 30， 50 μmol/L） on the morphology of human hepatoma cells （HepG2， Bel-7402）， human lung cancer cells （A549） and human colon cancer cells （HCT-8） were observed， and the toxicity of TA and SPV to four tumor cells was monitored. The effects of TA and SPV on the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of phosphorylated protein kinase B （p-Akt） （Thr308， Ser473）， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， survivin， C/EBP-homologous protein （CHOP）， immunoglobulin binding protein （Bip） and microtubule-associated protein 1 light chain 3Ⅱ （LC3Ⅱ） in HCT-8 cells were detected. RESULTS After the intervention of TA and SPV， the volume reduction and nuclear shrinkage were founded in four tumor cells； the cell activity decreased to varying degrees， among which TA and SPV had the best inhibitory effect on HCT-8 cells. After the intervention of TA and SPV， the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells， the protein expressions of Bax， CHOP， Bip and LC3Ⅱ all increased to different degrees， while the protein expressions of p-Akt （Thr308， Ser473）， Bcl-2 and survivin in HCT-8 cells all decreased to different degrees. CONCLUSIONS TA and SPV have inhibitory effects on the above four tumor cells， and the inhibitory effect on HCT-8 cells is the best. The mechanism of their action on HCT-8 cells may be related to promoting apoptosis， activating endoplasmic reticulum stress and autophagy.

Objective:To investigate the therapeutic effects of internal fixation via the posterolateral combined posteromedial approach in the treatment of posterior pilon fracture (Klammer type Ⅲ).Methods:A retrospective study was performed to analyze the 69 posterior pilon fractures (Klammer type Ⅲ) which had been treated by internal fixation with hollow screws or a buttress plate at Department of Orthopaedic Trauma, Yantai Shan Hospital from January 2015 to January 2020. There were 36 males and 33 females with an age of (45.3±10.0) years and duration from injury to surgery of (6.0±1.5) d. They were assigned into 2 groups according to different surgical approaches. The observation group (41 cases) was treated through the posterolateral combined posteromedial approach while the control group (28 cases) through the posterolateral approach alone. The therapeutic effects were evaluated by comparing the 2 groups in terms of incision length, intraoperative bleeding, operation time, fracture union time, fracture reduction (evaluated by the Burrwell-Charnley radiological score), the ankle-hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS) and complications.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). All the 69 cases were followed up for (16.9±4.0) months, revealing primary healing of all the incisions and no vascular injury or wound infection. The incision length [(11.2±1.8) cm] and operation time [(76.0±6.6) min] in the observation group were significantly shorter than those in the control group [(12.4±1.9) cm and (79.7±6.8) min], the excellent and good rate of reduction in the observation group (97.6%, 40/41) was significantly higher than that in the control group (89.3%, 25/28), and the ankle-hindfoot scores of AOFAS at 1, 3 and 12 months after operation in the observation group [(78.4±5.6), (79.5±2.8) and (86.9±2.1)] were significantly higher than those in the control group [(75.2±5.5), (78.0±3.2) and (85.8±2.3)] (all P<0.05). There was no significant difference in the intraoperative bleeding between the 2 groups ( P>0.05). In the control group, 2 patients developed numbness in the dorsum of foot, which gradually disappeared after 3 months of treatment, but no other complications like persistent pain or flexor contracture within 1 year after operation. Conclusion:In the treatment of Klammer type Ⅲ posterior pilon fracture, the posterolateral combined posteromedial approach can result in satisfactory therapeutic effects, because the surgical approach can fully expose the fracture and facilitate better reduction.

Objective:To evaluate the value of chemoradiotherapy and surgery in cervical esophageal cancer (CEC).Methods:Data of 459 patients with CEC from 2004 to 2017 were collected and retrospectively analyzed from the surveillance, epidemiology, and end results (SEER) database of National Cancer Institute (US). All patients were divided into the chemoradiotherapy group ( n=379) and surgery group ( n=80) according to the treatment methods. Survival analysis was performed by Kaplan-Meier method and survival curve was drawn. Multivariate survival analysis was conducted by Cox proportional hazards regression model. The death rate of different causes between two groups was calculated by cumulative incidence function (CIF). The differences of death rate between two groups were evaluated by Fine-Gray competing risk model. By analyzing the clinical characteristics and survival of CEC patients, the overall survival (OS) was compared between the surgery and chemoradiotherapy groups. Results:The 2- and 5-year survival rates in the chemoradiotherapy group were 43.1% and 22.4%, while those of the surgical group were 46.8% and 26.0%, respectively. No significant difference was observed in the OS between the chemoradiotherapy and surgery groups ( P=0.750). Cox multivariate analysis showed that treatment (surgery group vs. chemoradiotherapy group) was not an independent prognostic factor for OS. Based on the results of competing risk analysis, the risk of esophageal cancer-specific death in the chemoradiotherapy group was higher than that in the surgery group, and the difference was statistically significant between two groups ( P<0.001). The risk of other cause-specific death in the chemoradiotherapy group was lower than that in the surgery group ( P<0.001). The proportion of patients who died of oral, oropharyngeal, hypopharyngeal and laryngeal diseases in the surgery group was significantly higher than that in the chemoradiotherapy group(all P<0.001). Conclusions:No significant difference is observed in the OS of CEC patients treated with chemoradiotherapy or surgery. In the surgery group, the risk of esophageal cancer-specific death is lower, whereas the risk of other cause-specific death is higher compared with those in the chemoradiotherapy group.

Purpose: We aimed to investigate the mechanism of phenotypic transformation of corporal cavernosum smooth muscle cells (CCSMCs) under hypoxic conditions in vitro. Materials and Methods: In this study, a hypoxia model was established using cobalt chloride (CoCl2). CCSMCs were treated with different concentrations of CoCl2 for varying time periods, and cell viability was assessed. Hypoxia-inducible factor-1α (HIF-1α), myocardin (Myocd) and phenotypic markers were detected in the CCSMCs. We also transfected the CCSMCs with si-HIF-1α and Ad-Myocd and evaluated the effects on phenotypic modulation of CCSMCs and the relationship between HIF-1α and Myocd was evaluated. Results: CoCl2 inhibited the viability of CCSMCs in a dose- and time-dependent manner, and treatment with 300 µM CoCl2 for 48 hours were the optimal conditions for establishing the hypoxia model. The results showed increased expression levels of HIF-1α and osteopontin and decreased Myocd, alpha-smooth muscle actin, and calponin levels in CCSMCs under hypoxia. HIF-1α knockdown reversed hypoxia-induced phenotypic transformation with elevated Myocd expression. Overexpression of Myocd also reversed the effect of hypoxia on the phenotypic switch, but did not affect HIF-1α expression. Conclusions Our findings showed that HIF-1α was involved in the effect of hypoxia induced by CoCl2 on CCSMC phenotypic modulation, and Myocd overexpression could inhibit this process. Thus, Myocd might be a potential therapeutic target for erectile dysfunction under hypoxia or HIF-1α activation.

Objective:To evaluate the diagnostic values of serum fibrinogen and D-dimer for periprosthetic joint infection (PJI).Methods:The medical records were retrospectively analyzed of the 175 patients who had undergone hip or knee revisions at Department of Joint Surgery, Affiliated Hospital to Qingdao University from August 2013 to June 2019. Of them, 59 were diagnosed as PJI (31 knees and 28 hips), including 33 males and 26 females with an age of (67.4±11.7) years and a body mass index (BMI) of (26.1±3.6) kg/m 2 while 116 as aseptic loosening (AL) (19 knees and 97 hips), including 67 males and 49 females with an age of (70.3±8.9) years and a BMI of (25.0±3.6) kg/m 2. The plasma levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), fibrinogen and D-dimer in the 2 groups were recorded and analyzed. The receiver operating characteristic curve (ROC) was used to calculate the sensitivity and specificity of each indicator; the diagnostic value for each indicator was calculated according to the area under the curve (AUC). Results:There was no significant difference between the PJI and AL groups in gender, age or BMI ( P>0.05), but there was a significant difference in the joint type ( P<0.05). Compared with the AL group, the PJI group had significantly higher levels of CRP, ESR, fibrinogen and D-dimer ( P<0.05). The AUCs for CRP, ESR, fibrinogen and D-dimer were 0.830, 0.850, 0.848 and 0.664, respectively. By the Youden index, the optimal predictive cutoffs for CRP, ESR, fibrinogen and D-dimer were 8.06 mg/L, 17.60 mm/h, 3.73 g/L and 685.00 ng/mL, giving sensitivities of 79.2%, 85.4%, 81.3% and 64.6% and specificities of 85.7%, 76.2%, 79.8% and 61.9%. Conclusions:The diagnostic value of serum fibrinogen may be high for PJI, similar to that of CRP or ESR. However, D-dimer may be of limited value for diagnosis of PJI.

Objective:To evaluate the curative effect of 3D printing titanium trabecular metal （TTM） acetabular cups in revision of total hip arthroplasty （THA）.Methods:A retrospective case series study was conducted on the clinical data of 24 patients （25 hips） undergoing THA revision in Affiliated Hospital of Qingdao University from May 2016 to September 2019. There were 10 males and 14 females，aged 43-78 years ［（63.0 ± 11.4） years］. According to Paprosky classification，5 patients（5 hips） were classified as type Ⅰ，9 patients （10 hips） as type ⅡA，5 patients （5 hips） as type ⅡB，1 patient （1 hip） as type ⅡC and 4 patients （4 hips） as type ⅢC. All patients used 3D printing TTM cups. Seven patients （7 hips） were combined with 3D printing TTM pads，and 3 patients （3 hips） were combined with structural bone grafts. The visual analogue scale （VAS），Harris score and quality of life Health Survey Scale （SF-36） score were evaluated before operation，3 months after operation，12 months after operation and at the last follow-up. The X-ray film of hip joint was performed to evaluate the upward movement distance of hip center of rotation （HCOR），limb-length discrepancy （LLD） before operation，1 day after operation and at the last follow-up. The position of cups and bone ingrowth were evaluated at the last follow-up. The postoperative complications were observed.Results:All patients were followed up for 14-54 months ［（34.2 ± 9.3）months］. The VAS was （4.0 ± 0.7）points，（2.3 ± 0.8）points and （0.9 ± 0.2）points at postoperative 3 months，12 months and the last follow-up，significantly lower than that preoperatively ［（6.1 ± 1.0）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative VAS at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The Harris score was （64.6 ± 5.3）points，（80.5 ± 3.7）points，and （90.3 ± 3.6）points at postoperative 3 months，12 months and the last follow-up，significantly higher than that preoperatively ［（38.8 ± 6.2）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative Harris score at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The SF-36 score was （556.3 ± 21.9）points，（711.6 ± 15.9）points and （752.8 ± 23.0）points at postoperative 3 months，12 months and the last follow-up，significantly higher than that preoperatively ［（326.3 ± 30.7）points］ （ P < 0.05）. A statistically significant difference was found in the pairwise comparisons of postoperative SF-36 score at postoperative 3 months，12 months and the last follow-up（ P < 0.05）. The upward movement distance of HCOR was （13.5 ± 2.6）mm and （13.6 ± 2.6）mm on the first day after operation and at the last follow-up，significantly lower than that preoperatively ［（34.1 ± 3.5）mm］ （ P < 0.05）. The LLD was （6.2 ± 1.8）mm and （5.3 ± 1.5）mm on the first day after operation and at the last follow-up，significantly lower than that preoperatively ［（31.6 ± 5.2）mm］ （ P < 0.05）. All the cups were stable. Good bone ingrowth was found in 23 hips. There was no postoperative complication except for one patient with poor wound healing. Conclusion:For THA revision，the use of 3D printing TTM cups can effectively relieve pain，improve hip function，provide satisfactory biological fixation and and reduce complications.

Objective:To evaluate the diagnostic value of platelet count(PC),PC to mean platelet volume(MPV) ratio(PC/MPV) and plateletcrit(PCT) in chronic periprosthetic joint infection(PJI).Methods:The medical records of 159 patients who underwent hip or knee revisions at Department of Joint Surgery, Affiliated Hospital of Qingdao University from August 2013 to June 2019 were retrospectively reviewed. There were 51 patients(26 knees and 25 hips) in the PJI group,which included 28 males and 23 females,aged (68.0±11.8)years (range:32 to 84 years)with a body mass index(BMI)of (26.1±3.6) kg/m2.There were 116 patients(19 knees and 97 hips) in the aseptic loosening(AL) group,including 67 males and 49 females,aged (70.3±8.9)years(range:49 to 89 years)with a BMI of (25.0±3.6)kg/m2.The plasma C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),PC,MPV,PC/MPV and PCT levels of the two groups were recorded and analyzed. Receiver operating characteristic curve was used to calculate the sensitivity and specificity of each biomarker,expect for MPV,and the diagnostic value of each biomarker was compared according to the area under the curve(AUC).Independent-sample t test or Mann-Whitney U test were used for comparison between groups. Results:Compared with AL group,AJI group had significantly higher levels of CRP,ESR,PC,PC/MPV and PCT(all P<0.05),but lower level of MPV ( P<0.05).The AUCs for CRP,ESR,PC,PC/MPV and PCT were 0.820, 0.829, 0.689, 0.668 and 0.676,respectively. Based on the Youden index,the optimal predictive cutoff for CRP was 11.12 mg/L,with a sensitivity of 74.4% and a specificity of 87.1%.The optimal predictive cutoff for ESR was 17.60 mm/1 h,with a sensitivity of 81.4% and a specificity of 75.3%.The optimal predictive cutoff for PC was 243.00×10 9/L,with a sensitivity of 60.6% and a specificity of 71.8%.The optimal predictive cutoff for PC/MPV was 24.95,the sensitivity was 58.1% and the specificity was 74.1%.And the optimal predictive cutoff for PCT was 0.24%,with a sensitivity of 69.8% and a specificity of 63.5%. Conclusion:PC,PC to MPV ratio and PCT are of limited value to diagnose PJI.

Objective:To evaluate the diagnostic value of platelet count(PC),PC to mean platelet volume(MPV) ratio(PC/MPV) and plateletcrit(PCT) in chronic periprosthetic joint infection(PJI).Methods:The medical records of 159 patients who underwent hip or knee revisions at Department of Joint Surgery, Affiliated Hospital of Qingdao University from August 2013 to June 2019 were retrospectively reviewed. There were 51 patients(26 knees and 25 hips) in the PJI group,which included 28 males and 23 females,aged (68.0±11.8)years (range:32 to 84 years)with a body mass index(BMI)of (26.1±3.6) kg/m2.There were 116 patients(19 knees and 97 hips) in the aseptic loosening(AL) group,including 67 males and 49 females,aged (70.3±8.9)years(range:49 to 89 years)with a BMI of (25.0±3.6)kg/m2.The plasma C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),PC,MPV,PC/MPV and PCT levels of the two groups were recorded and analyzed. Receiver operating characteristic curve was used to calculate the sensitivity and specificity of each biomarker,expect for MPV,and the diagnostic value of each biomarker was compared according to the area under the curve(AUC).Independent-sample t test or Mann-Whitney U test were used for comparison between groups. Results:Compared with AL group,AJI group had significantly higher levels of CRP,ESR,PC,PC/MPV and PCT(all P<0.05),but lower level of MPV ( P<0.05).The AUCs for CRP,ESR,PC,PC/MPV and PCT were 0.820, 0.829, 0.689, 0.668 and 0.676,respectively. Based on the Youden index,the optimal predictive cutoff for CRP was 11.12 mg/L,with a sensitivity of 74.4% and a specificity of 87.1%.The optimal predictive cutoff for ESR was 17.60 mm/1 h,with a sensitivity of 81.4% and a specificity of 75.3%.The optimal predictive cutoff for PC was 243.00×10 9/L,with a sensitivity of 60.6% and a specificity of 71.8%.The optimal predictive cutoff for PC/MPV was 24.95,the sensitivity was 58.1% and the specificity was 74.1%.And the optimal predictive cutoff for PCT was 0.24%,with a sensitivity of 69.8% and a specificity of 63.5%. Conclusion:PC,PC to MPV ratio and PCT are of limited value to diagnose PJI.

Objective To investigate the prevalence of fluorosis and related control measures on drinking water type of endemic fluorosis in China.Methods According to the national program"Surveillance Scheme of Drinking-Water-Borne Endemic Fluorosis",136 counties were selected in 29 provinces,autonomous regions and municipalities.Three epidemic villages were randomly selected as fixed monitoring sites in each county.Dental fluorosis of all the children aged 8-12 living in the villages under the monitoring program,was identified under the ariteria from "Diagnosis of dental fluorosis"(WS/T 208-2011).Operating conditions and contents of fluoride in all the'water-improved projects' were investigated.Contents of fluoride in drinking water were tested in villages without the 'water-improved projects'."Standard Test Method for Drinking Water" (GB/T 5750.5-2006) was used to detect the water fluoride.Results The overall prevalence of dental fluorosis among children aged 8-12 in all the villages under monitor program,was 28.58％ (7 950/27 817),with the dental fluorosis index (DFI) as 0.58.Among them,the prevalence was 22.28％ (3 917/17 583) and DFI was 0.44 in the' water-improved projects' villages that under normal operation and with qualified fluoride contents.The prevalence appeared as 38.74％ (1 926/4 971) with DFI as 0.84 in those villages with 'water-improved projects' but mal-operated or with excessive fluoride.The prevalence was 40.03％ (2 107/5 263),and DFI was 0.81 in those villages without'water-improved projects'.The prevalence rates of dental fluorosis in children from the three types of endemic areas were significantly different.For 'water-improved projects',the normal opration rate was 93.77％ (286/305) and the qualification rate of fluoride content was 76.77％ (228/297).Conclusions Dental fluorosis in children living in the drinking-water-born endemic fluorosis areas was on the edge of epidemics in China.Effective improvement on the quality of drinking water can significantly reduce the severity of dental fluorosis in children.The rate of proper operation on 'water-improved projects' was near to 95％ in the endemic area.However,rate that met the criteria on qualified fluoride contents of these projects was still below 80％.

Objective To investigate the implementation effects of control measures and the dynamic prevalence of drinking-water-borne endemic fluorosis,in order to provide a scientific basis for developing control measures against the disease.Methods According to the Surveillance Scheme of Drinking-Water-Borne Endemic Fluorosis,137 counties were selected as monitoring counties in national 27 provinces (autonomous regions,municipalities) and Xin Jiang Production and Construction Corps.Three epidemic villages were selected as fixed monitoring villages in each county.The operating condition and the fluoride level of water improvement projects were investigated in the villages with water improvement projects.The fluoride level of drinking water was tested in the villages without water improvement projects.The fluoride content in drinking water was detected by the Standard Test Method for Drinking Water (GB/T 5750.5-2006).The preyalence of dental fluorosis of all children aged 8-12 were surveyed,and dental fluorosis was diagnosed by Clinical Diagnostic Criteria of Dental Fluorosis (WS/T 208-2011).Results ①427 villages were surveyed in 2012,among which the villages with water improvement projects accounted for 75.88％ (324/427).② 299 water improvement projects were investigated.Normal operation rates of those were 89.30％ (267/299).The qualified rate of fluoride content was 75.78％ (219/289).Totally 103 villages without water improvement projects were monitored.Villages with fluoride level ＞ 1.2-2.0 mg/L,＞ 2.0-4.0 mg/L and ＞ 4.0 mg/L accounted for 48.54％ (50/103),29.13％ (30/103) and 9.71％ (10/103),respectively.(③The national detection rate of dental fluorosis and community fluorosis index (CFI) in children aged 8-12 were 31.24％ (8 677/27 773) and 0.66.The detection rate and CFI of the villages with normal operation projects and the qualified fluoride content were 24.04％ (3 999/16 634) and 0.51.The detection rate and CFI of the villages with water improvement projects under abnormal operation or excessive fluoride were 40.77％ (1 654/4 057) and 0.91.The rate and CFI of the villages without water improvement projects were 42.70％ (3 024/7 082) and 0.86.Conclusions The rate of water improvement projects of the whole country is lower.The national normal operation rate of water defluoridation improvement projects is below 90％,and the qualified rate of fluoride content in drinking water is below 80％.The prevalence of dental fluorosis in children aged 8-12 is very mild.