Objective:To investigate the association of urinary cadmium levels with peripheral leukocyte classification counts among middle-aged and older adults aged 40 to 89 years in selected areas of China.Methods:The research was based on the survey of the impact of soil quality of agricultural land on human health in typical areas conducted in 2019-2020. A total of 5 600 middle-aged and older adults aged 40 to 89 years were included by using a multi-stage stratified random sampling method. Baseline characteristics of the subjects were collected and physical examinations were performed. Random midstream urine was collected to measure urinary cadmium and urinary creatinine and fasting venous blood was collected to measure the leukocyte count, neutrophil count, lymphocyte count, monocyte count and eosinophil count. The linear mixed effect model was used to analyse the association of urinary cadmium levels with leukocyte classification counts, and the dose-response relationship between them was analyzed by using the restricted cubic spline (RCS) function.Results:The age of the subjects was (63.17±12.02) years; 2 851 (50.91%) were males; and the M ( Q 1, Q 3) of urinary creatinine-corrected urinary cadmium levels was 2.69 (1.52, 4.69) μg/g·creatinine. After adjusting for confounding factors, the results of linear mixed effects model analysis showed that for each 1-unit increase in urinary creatinine-corrected urinary cadmium level, the percentage change [% (95% CI)] of leukocyte count and lymphocyte count was -1.70% (-2.61%, -0.79%) and -1.57% (-2.86%, -0.26%), respectively. RCS function showed a negative linear relationship between urinary creatinine-corrected urinary cadmium levels and leukocyte counts and lymphocyte counts, respectively (all Pnon-linear>0.05). Conclusion:Urinary cadmium levels are negatively associated with leukocyte count and lymphocyte count among middle-aged and older adults aged 40 to 89 years in selected areas of China.

Objective:To investigate the association of urinary cadmium levels with peripheral leukocyte classification counts among middle-aged and older adults aged 40 to 89 years in selected areas of China.Methods:The research was based on the survey of the impact of soil quality of agricultural land on human health in typical areas conducted in 2019-2020. A total of 5 600 middle-aged and older adults aged 40 to 89 years were included by using a multi-stage stratified random sampling method. Baseline characteristics of the subjects were collected and physical examinations were performed. Random midstream urine was collected to measure urinary cadmium and urinary creatinine and fasting venous blood was collected to measure the leukocyte count, neutrophil count, lymphocyte count, monocyte count and eosinophil count. The linear mixed effect model was used to analyse the association of urinary cadmium levels with leukocyte classification counts, and the dose-response relationship between them was analyzed by using the restricted cubic spline (RCS) function.Results:The age of the subjects was (63.17±12.02) years; 2 851 (50.91%) were males; and the M ( Q 1, Q 3) of urinary creatinine-corrected urinary cadmium levels was 2.69 (1.52, 4.69) μg/g·creatinine. After adjusting for confounding factors, the results of linear mixed effects model analysis showed that for each 1-unit increase in urinary creatinine-corrected urinary cadmium level, the percentage change [% (95% CI)] of leukocyte count and lymphocyte count was -1.70% (-2.61%, -0.79%) and -1.57% (-2.86%, -0.26%), respectively. RCS function showed a negative linear relationship between urinary creatinine-corrected urinary cadmium levels and leukocyte counts and lymphocyte counts, respectively (all Pnon-linear>0.05). Conclusion:Urinary cadmium levels are negatively associated with leukocyte count and lymphocyte count among middle-aged and older adults aged 40 to 89 years in selected areas of China.

Objective:To investigate the effect of gap junction protein Cx43 inhibitor carbenoxolone (CBX) on cognitive function and its possible mechanism in epileptic rats.Methods:One hundred and twenty Wistar rats were randomly divided into sham-operated group, epilepsy group, epilepsy+solvent group, and epilepsy+CBX group ( n=30). The models of temporal lobe epilepsy in the later three groups were prepared by injection of kainic acid in the hippocampus. Intraperitoneal injection of CBX (20 mg/kg) or equal amount of normal saline were given to the rats in the epilepsy+CBX group and epilepsy+solvent group 30 min before modeling. Western blotting was used to detect the protein expressions of phosphorylated (p)-Cx43 and microtubule associated protein light chain 3 (LC3) in the hippocampus 6, 12, and 24 h after modeling; the protein localization of p-Cx43 and LC3 in the hippocampus and optical density of their positive cells were detected by immunohistochemistry 24 h after modeling; the learning and memory abilities of rats were tested by Morris water maze experiment 30 d after modeling. Results:Western blotting results showed that as compared with those in the sham-operated group, p-CX43 and LC3 protein expressions in the hippocampal CA3 regions of epilepsy group and epilepsy+solvent group were significantly increased at 6, 12 and 24 h after modeling ( P<0.05); as compared with the epilepsy group and epilepsy+solvent group, the epilepsy+CBX group had statistically decreased p-CX43 and LC3 protein expressions in the hippocampal CA3 regions at each time point ( P<0.05). Immunohistochemical staining showed that p-CX43 was localized at the cell membrane and cytoplasm of hippocampal astrocytes; LC3 was located at the cytoplasm of hippocampal neurons. As compared with those in the sham-operated group, the optical density values of p-CX43 and LC3 positive cells in hippocampal CA3 regions of epilepsy group and epilepsy+solvent group were increased ( P<0.05). As compared with those in the epilepsy group and the epilepsy+solvent group, the optical density values of p-CX43 and LC3 positive cells in the hippocampal CA3 regions of the epilepsy+CBX group were significantly decreased ( P<0.05). Morris water maze test results showed that as compared with that in the sham-operated group, the escape latency in the epilepsy group and epilepsy+solvent group was significantly prolonged ( P<0.05); as compared with that in the epilepsy group and epilepsy+solvent group, the latency in the epilepsy+CBX group was significantly shortened ( P<0.05). Conclusion:CBX can weaken the neuronal autophagy and reduce the damage to cognitive function by inhibiting the p-Cx43 protein expression in the astrocytes of the hippocampal CA3 regions.

Objective To evaluate the validity and security of our self-designed bone marrow stem cell Screen-Enrich-Combine(-Biomaterials) Circulating System(SECCS) for bone non-union of limbs. Methods From May 2013 to December 2015, 24 patients with limb non-union were treated at our de-partment. They were aged from 20 to 65 years (mean, 42.8 years). Their non-union involved femur in 17 cases, tibia in 4, radius in one, humerus in one and fibula in one. In surgery, 80 mL bone marrow blood was aspirated from the anterior superior iliac spine for rapid preparation of bone substitute(β-TCP)composite with bone marrow stem cells by SECCS which was then implanted at the non-union locations. The bone marrow blood was collected before and after enrichment for stem cell counts. The bone union, clinical union time and related com-plications were evaluated by follow-up X-rays at 1, 3, 6 and 9 months after surgery. Results Each collection of bone marrow blood took 13.5 minutes on average. The enrichment rate of stem cells was 81.2%. 11,751 ± 1,011 stem cells were implanted per patient on average. All the patients were followed up for 9 to 48 months (mean, 19.2 months). Twenty-two patients acquired clinical union 9 months after operation and the other 2 suffered from malunion due to insufficient bone implant, yielding a union rate of 91.6% and an average union time of 6.5 months. No graft infection or internal fixation failure occurred and no severe complications hap-pened at the donor or implant sites.Conclusion The bioactive bone substitute manufactured by our self-designed SECCS can be used as a novel therapy for limb non-union, and this device is moreover charac-terized with convenience, limited invasion and satisfactory osteogenesis so that complications of autologous bone transplantation can be avoided.

Objective To introduce a new method for preparation of bioactive β-tricalcium phosphate (β-TCP) by rapid stem cell screen-and-enrich-and-combine circulating system (SECCS) and evaluate its efficacy in the treatment of fresh fractures and bone defects.Methods Twenty-two patients with fresh fracture and bone defects were treated with SECCS from July 2013 to April 2016.They were 16 males and 6 females with an average age of 52.2 years (from 27 to 81 years).There were 15 tibial plateau fractures and 7 calcaneal fractures.The average size of bone defects was 12.5 mL.Bioactive β-TCP was prepared by SECCS intraoperatively and implanted back immediately into the bone defects.Radiographic examination,Lysholm knee scoring and Maryland foot scoring were used for assessment of curative efficacy.Results The 22 patients were followed up for an average of 25.7 months (from 12 to 46 months).By SECCS,the enrichment efficiency of bone marrow stromal cells (BMSCs) reached up to 82.4％ and the cell viability was not affected.The tibial plateau fractures were re-transplanted with 13,381.3 BMSCs on average and healed after an average of 8.9 weeks (from 6 to 15 weeks).The Lysholm knee scores at one year postoperatively averaged 93.6 points (from 84 to 100 points),yielding 10 excellent cases,4 good cases and one fair case.The calcaneal fractures were implanted back with 16,677.7 BMSCs on average and healed after an average of 9.4 weeks (from 8 to 13 weeks).The average Maryland foot score at one year after operation was 93.6 points (from 85 to 98 points),yielding 6 excellent cases and one good case.Conclusion Bioactive materials prepared by SECCS are good bone grafts for fresh fractures and bone defects.

Objective To evaluate the efficiency of our self-designed enriched filtrator of stem cells in screening and collecting enriched human bone marrow stem cells (BMSCs) for rapid construction of bioactive materials.Methods Bone marrow blood from 32 patients was circulated in our self-designed enriched filtratorand filtered through beta-tricalcium phosphate(β-TCP).The numbers of BMSCs were counted pre-and post-filtration to assess the enrichment efficiency.The numbers of nucleated cells,red blood cells and platelets were compared pre-and post-filtration to assess the effects of the enriched filtrator of stem cells on blood cells.Bacteriological examinations were performed to test the safety of the device.Scanning electron microscopy was carried out to observe the adhesion of BMSCs on β-TCP.Results There was a significant difference between the pre-fihration colony number of BMSCs cultured (232.8 ± 128.6/mL) and the post-filtration one (37.8 ± 34.9/mL) (t =10.743,P ＜ 0.001).The efficiency of BMSCs enrichment of the device was 85.6％ ±8.4％ (from 63.8％ to 97.1％).There were no significant differences between pre-filtration and post-filtration in the numbers of nucleated cells [(16.8 ± 5.4) × 109/mL versus (16.3 ±5.0) ×l09/mL],red blood cells [(3.9±0.6) ×1012/mL versus (3.8±0.6) ×1012/mL] and platelets [(120.4 ± 54.6) × 109/mL versus (123.0 ± 51.8) × 109/mL] (P ＞ 0.05).All the bacteriological examinations of the bone marrow samples were negative.After in vitro osteogenic induction culture,formation of numerous BMSCs colonies was observed on the β-TCP by scanning electron microscopy.Conclusion Since our self-designed enriched filtrator can efficiently collect enriched BMSCs and combine them with β-TCP simultaneously,the device is capable of rapid and safe construction of bioactive materials.

Objective To build a multi‐level hierarchical structure model of the influencing factors for hierarchical medical system ,to identify the role relationship between all the factors and transmission pathways ,and to recommend on developing China′s hierarchical medical system . Methods Thirty influencing factors were identified in a screening based on literature review for the hierarchical medical system .On such basis ,16 influencing factors were identified by three health policy experts ,which affect operations of the current system .Interpretative structural modeling was called into play in the end to analyze the hierarchy relationship between various influencing factors and the conduction loops .Results There exist among the 16 factors a 3‐level hierarchical structureand two conduction loops .The factor directly limiting the hierarchical medical system is two‐way referral,and most internal core drivers arehuman resources development and governance mechanism.By means of self‐growth and external constraints ,they exert their influence on the operation of hierarchical medical system .Conclusions There are interactive hierarchical effects among the factors ,merging into three node elements of functional role,inter‐entity relationshipand patient participation.

Objective To investigate the expression of aquaporin-8(AQP8)and apoptosis associated bcl-2 protein in human cervical carcinoma and their relationship.Methods The expression of AOP8 and bcl-2 protein in 74 cases of cervical carcinoma (46 cases of squamous-cell carcinoma of the uterine cervix,28 cases of adenocarcinoma of the uterine cervix),34 cases of cervical intraepithelial neoplasia (CIN)and 15 cases of normal cervices were detected by immunohistochemical technique,and their clinical significance were analyzed.Results The expression of AQP8 and bcl-2 protein were detected in intracytoplasm of atypia cells in CIN.squamous-cell carcinoma and adenocarcinoma of the uterine cervix.The positive rates of AQP8 and bcl-2 in squamous-cell carcinoma.adenocarcinoma,CIN and normal cervical epithelium were 98%,74%;61%,71%;71%,53%;53%,20%respectively.There were significant differences between squamous-cell carcinoma of the uterine cervix and other groups in AQP8(P＜0.01),but no significant differences were found in any other groups.There were significant differences between squamous-cell carcinoma of the uterine cervix and CIN or normal cervical epithelium in bcl-2.so were between adenocarcinoma of the uterine cervix.The expression of AQP8 was positively correlated with bcl-2 in human cervical carcinoma(rs=0.463,P=0.000).Conclusions There is a close relationship between high expression of AQP8 and development of human cervical carcinoma.The expression of AQP8 protein is positively correlated with bcl-2 protein in human cervical carcinoma. AQP8 protein may have anti-apoptosis function.although the detailed mechanism in human cervical carcinoma remaias to be clarified.

Objective To found sensitive and reliable method to identify trophoblastic tumor cells in the peripheral blood of the patients suffered from gestational trophoblastic tumor.Methods Given numbers of JAR cell from ten to million were mixed into 10ml non pregnant peripheral blood as a model. Detection of ? hCG mRNA with fluorescence quantitative reverse transcription polymerase chain reaction (FQ RT PCR) and then estimation of the numbers of tumor cell in the blood. Nine cases of peripheral blood were collected from the pretreatment patients of gestational trophoblastic tumor to assay ? hCG mRNA with FQ RT PCR, then to estimate the numbers of tumor cell in the circulation blood. Results FQ RT PCR could detect ? hCG mRNA when ≥10 2 JAR cells were mixed into 10ml non pregnant peripheral blood. Four cases of bloods had been detected ? hCG mRNA expression in 9 cases of gestational trophoblastic tumor, and the numbers of tumor cell from 10 4 to 10 7 per 10ml blood. Conclusion FQ RT PCR of which primers and probe are designed for ? hCG had been proved to be very sensitive detection means, it could be used to detect gestational trophoblastic tumor cells from patient preipheral blood; With FQ RT PCR the tumor cells had been detected in some patients of gestational trophoblastic tumor.

Objectives To investigate HPV infection and its correlati on with the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibito r of metalloproteinase-1 (TIMP-1) in vulvar squamous cell carcinoma (VSCC). Meth ods HPV6/11 and 16/18 DNA were detected using in situ hybridization in 26 case s of VSCC, 21 cases of vulvar intraepithelial neoplasia (VIN) and 10 specimens o f normal skin. The expression of MMP-9 and TIMP-1 was measured by immunohistoche mical method in these three groups. Results The infection rates of HPV6/11 and 16/18 in VSCC and VIN were 69.23% (18/26) and 38.46% (10/26), 42.86% (9/21) and 28.57% (6/21) respectively, and no HPV6/11 or 16/18 DNA was discovered in norma l skin epidermis. The expression rates of MMP-9 and TIMP-1 in VSCC, VIN and norm al skin epidermis were 92.31%(24/26) and 76.92% (20/26), 90.45% (19/21) and 80.9 5% (17/21), and 80.00% (8/10) and 50.00% (5/10) respectively. MMP-9 expression in HPV positive lesions was higher than that in HPV negative lesions, but no sig nificant difference of TIMP-1 expression was observed in HPV positive and negati ve lesions. Conclusions HPV infection may play a role in the development of VS CC. The higher expression of MMP-9 comparing with that of TIMP-1 may be an early marker of tumor progression in VSCC, and HPV infection may facilitate the invas ion and metastasis of VSCC.