Objective·To study the effect of neodymium-doped yttrium aluminum perovskite(Nd:YAP)laser biostimulation on the proliferation and osteogenic differentiation of human periodontal ligament cells(hPDLCs)and its possible mechanism.Methods·Five premolars removed for orthodontic reasons were collected from Changzhou Stomatological Hospital,and the periodontal ligament tissues from the middle 1/3 of the roots were taken to culture hPDLCs in vitro.The cells were irradiated with the biostimulation function[G(-)mode]of the Nd:YAP laser.According to the irradiation time,the cells were divided into a control group(without laser irradiation),and groups irradiated for 5 s,10 s,15 s,20 s and 30 s.The cell counting kit-8(CCK-8)method was used to detect the proliferation of hPDLCs in each group.After osteogenic differentiation was induced,the alkaline phosphatase(ALP)content and activity level of the cells were detected using an ALP staining kit and an ALP activity detection kit.The calcium salt level of the cells was evaluated by alizarin red S staining and calcium quantitative analysis.The expression of genes and proteins related to the WNT/β-catenin signaling pathway,including dickkopf-related protein 1(DKK-1),β-catenin,and runt-related transcription factor 2(RUNX2),was analyzed by quantitative real-time PCR(qPCR)and Western blotting.Results·The results of CCK-8 showed that the proliferation level of cells in the 10 s,15 s,20 s,and 30 s groups was enhanced from 3 d after irradiation(all P<0.05).After induction of osteogenic differentiation,ALP content,activity,and calcium salt level in the laser irradiation groups increased with the extension of irradiation time(all P<0.05).The results of qPCR and Western blotting analysis showed that the expression levels of the DKK-1 gene and protein in the laser irradiation groups decreased with the extension of irradiation time.However,the expression levels of β-catenin and RUNX2 genes and proteins increased significantly with the extension of irradiation time;there were statistically significant differences between the 15 s,20 s,and 30 s groups and the control group(all P<0.05).Conclusion·Nd:YAP laser biostimulation may promote proliferation and osteogenic differentiation of hPDLCs through WNT/β-catenin signaling pathway.

Objective To investigate the changes in eosinophil counts and the activation of microglial cells in the brain tissues of mice at different stages of Angiostrongylus cantonensis infection, and to examine the role of microglia in regulating the progression of angiostrongyliasis and unravel the possible molecular mechanisms. Methods Fifty BALB/c mice were randomly divided into the control group and the 7-d, 14-d, 21-day and 25-d infection groups, of 10 mice in each group. All mice in infection groups were infected with 30 stage III A. cantonensis larvae by gavage, and animals in the control group was given an equal amount of physiological saline. Five mice were collected from each of infection groups on days 7, 14, 21 d and 25 d post-infection, and 5 mice were collected from the control group on the day of oral gavage. The general and focal functional impairment was scored using the Clark scoring method to assess the degree of mouse neurological impairment. Five mice from each of infection groups were sacrificed on days 7, 14, 21 d and 25 d post-infection, and 5 mice from the control group were sacrificed on the day of oral gavage. Mouse brain tissues were sampled, and the pathological changes of brain tissues were dynamically observed using hematoxylin and eosin (HE) staining. Immunofluorescence staining with eosinophilic cationic protein (ECP) and ionized calcium binding adaptor molecule 1 (Iba1) was used to assess the degree of eosinophil infiltration and the counts of microglial cells in mouse brain tissues in each group, and the morphological parameters of microglial cells (skeleton analysis and fractal analysis) were quantified by using Image J software to determine the morphological changes of microglial cells. In addition, the expression of M1 microglia markers Fcγ receptor III (Fcgr3), Fcγ receptor IIb (Fcgr2b) and CD86 antigen (Cd86), M2 microglia markers Arginase 1 (Arg1), macrophage mannose receptor C-type 1 (Mrc1), chitinase-like 3 (Chil3), and phagocytosis genes myeloid cell triggering receptor expressed on myeloid cells 2 (Trem2), CD68 antigen (Cd68), and apolipoprotein E (Apoe) was quantified using real-time quantitative reverse transcription PCR (RT-qPCR) assay in the mouse cerebral cortex of mice post-infection. Results A large number of A. cantonensis larvae were seen on the mouse meninges surface post-infection, and many neuronal nuclei were crumpled and deeply stained, with a large number of bleeding points in the meninges. The median Clark scores of mouse general functional impairment were 0 (interquartile range, 0), 0 (interquartile range, 0.5), 6 (interquartile range, 1.0), 14 (interquartile range, 8.5) points and 20 (interquartile range, 9.0) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.45, P < 0.01), and the median Clark scores of mouse focal functional impairment were 0 (interquartile range, 0), 2 (interquartile range, 2.5), 7 (interquartile range, 3.0), 18 (interquartile range, 5.0) points and 25 (interquartile range, 6.5) points in the control group and the 7-d, 14-d, 21-d and 25-d groups, respectively (H = 22.72, P < 0.01). The mean scores of mice general and focal functional impairment were all higher in the infection groups than in the control group (all P values < 0.05). Immunofluorescence staining showed a significant difference in the eosinophil counts in mouse brain tissues among the five groups (F = 40.05, P < 0.000 1), and the eosinophil counts were significantly higher in mouse brain tissues in the 14-d (3.08 ± 0.78) and 21-d infection groups (5.97 ± 1.37) than in the control group (1.00 ± 0.28) (both P values < 0.05). Semi-quantitative analysis of microglia immunofluorescence showed a significant difference in the counts of microglial cells among the five groups (F = 17.66, P < 0.000 1), and higher Iba1 levels were detected in mouse brain tissues in 14-d (5.75 ± 1.28), 21-d (6.23 ± 1.89) and 25-d infection groups (3.70 ± 1.30) than in the control group (1.00 ± 0.30) (all P values < 0.05). Skeleton and fractal analyses showed that the branch length [(162.04 ± 34.10) μm vs. (395.37 ± 64.11) μm; t = 5.566, P < 0.05] and fractal dimension of microglial cells (1.30 ± 0.01 vs. 1.41 ± 0.03; t = 5.266, P < 0.05) were reduced in mouse brain tissues in the 21-d infection group relative to the control group. In addition, there were significant differences among the 5 groups in terms of M1 and M2 microglia markers Fcgr3 (F = 48.34, P < 0.05), Fcgr2b (F = 55.46, P < 0.05), Cd86 (F = 24.44, P < 0.05), Arg1 (F = 31.18, P < 0.05), Mrc1 (F = 15.42, P < 0.05) and Chil3 (F = 24.41, P < 0.05), as well as phagocytosis markers Trem2 (F = 21.19, P < 0.05), Cd68 (F = 43.95, P < 0.05) and Apoe (F = 7.12, P < 0.05) in mice brain tissues. Conclusions A. cantonensis infections may induce severe pathological injuries in mouse brain tissues that are characterized by massive eosinophil infiltration and persistent activation of microglia cells, thereby resulting in progressive deterioration of neurological functions.

Objective To investigate the correlation between cognitive impairment and depression in elderly patients with type 2 diabetes mellitus(T2DM).Methods Based on China Health and Retirement Longitudinal Study in 2011,totally 521 elderly T2DM patients were enrolled,and according to the results of minimum mental state examination(MMSE)and their education lev-els,they were divided into a cognitively impaired group(437 cases)and a cognitively normal group(84 cases).Center for ED Epidemiological Survey Depression Scale was used to assess the depression symptoms.The correlation between cognitive impairment and depression was analyzed.Results Compared with the cognitively normal group,the cognitively impaired group had signifi-cantly advanced age(71.98±5.29 year vs 69.42±3.98 year,P=0.000),larger proportion of de-pression(60.6％vs 35.7％,P=0.000),and higher C-reactive protein level(5.09±12.80 mg/L vs 2.25±2.43 mg/L,P=0.000),and obviously lower ratios of being married(72.1％vs 86.9％,P=0.001)and having cardiovascular disease(20.1％vs 32.1％,P=0.010),and decreased estimated glomerular filtration rate[77.15±15.88 ml/(min·1.73 m2)vs 81.91±13.55 ml/(min·1.73 m2),P=0.001].Multivariate logistic regression analysis showed that cognitive impairment was an independent risk factor for the development of depression in elderly T2DM patients(OR=3.44,95％CI:1.89-6.27,P＜0.01).ROC curve analysis indicated that the AUC value of MMSE score in predicting depression in elderly T2DM patients was 0.669(95％CI:0.626-0.709,P＜0.01).The direct effect of cognitive impairment and the mediating effect of loneliness on depression in elderly T2DM patients accounted for 72.22％and 27.78％of the total effect,respectively.Conclu-sion Cognitive impairment is associated with the presence of depression in elderly T2DM pa-tients,and loneliness plays a mediating role.

Objective To investigate the correlation between cognitive impairment and depression in elderly patients with type 2 diabetes mellitus(T2DM).Methods Based on China Health and Retirement Longitudinal Study in 2011,totally 521 elderly T2DM patients were enrolled,and according to the results of minimum mental state examination(MMSE)and their education lev-els,they were divided into a cognitively impaired group(437 cases)and a cognitively normal group(84 cases).Center for ED Epidemiological Survey Depression Scale was used to assess the depression symptoms.The correlation between cognitive impairment and depression was analyzed.Results Compared with the cognitively normal group,the cognitively impaired group had signifi-cantly advanced age(71.98±5.29 year vs 69.42±3.98 year,P=0.000),larger proportion of de-pression(60.6％vs 35.7％,P=0.000),and higher C-reactive protein level(5.09±12.80 mg/L vs 2.25±2.43 mg/L,P=0.000),and obviously lower ratios of being married(72.1％vs 86.9％,P=0.001)and having cardiovascular disease(20.1％vs 32.1％,P=0.010),and decreased estimated glomerular filtration rate[77.15±15.88 ml/(min·1.73 m2)vs 81.91±13.55 ml/(min·1.73 m2),P=0.001].Multivariate logistic regression analysis showed that cognitive impairment was an independent risk factor for the development of depression in elderly T2DM patients(OR=3.44,95％CI:1.89-6.27,P＜0.01).ROC curve analysis indicated that the AUC value of MMSE score in predicting depression in elderly T2DM patients was 0.669(95％CI:0.626-0.709,P＜0.01).The direct effect of cognitive impairment and the mediating effect of loneliness on depression in elderly T2DM patients accounted for 72.22％and 27.78％of the total effect,respectively.Conclu-sion Cognitive impairment is associated with the presence of depression in elderly T2DM pa-tients,and loneliness plays a mediating role.

Objective·To study the effect of neodymium-doped yttrium aluminum perovskite(Nd:YAP)laser biostimulation on the proliferation and osteogenic differentiation of human periodontal ligament cells(hPDLCs)and its possible mechanism.Methods·Five premolars removed for orthodontic reasons were collected from Changzhou Stomatological Hospital,and the periodontal ligament tissues from the middle 1/3 of the roots were taken to culture hPDLCs in vitro.The cells were irradiated with the biostimulation function[G(-)mode]of the Nd:YAP laser.According to the irradiation time,the cells were divided into a control group(without laser irradiation),and groups irradiated for 5 s,10 s,15 s,20 s and 30 s.The cell counting kit-8(CCK-8)method was used to detect the proliferation of hPDLCs in each group.After osteogenic differentiation was induced,the alkaline phosphatase(ALP)content and activity level of the cells were detected using an ALP staining kit and an ALP activity detection kit.The calcium salt level of the cells was evaluated by alizarin red S staining and calcium quantitative analysis.The expression of genes and proteins related to the WNT/β-catenin signaling pathway,including dickkopf-related protein 1(DKK-1),β-catenin,and runt-related transcription factor 2(RUNX2),was analyzed by quantitative real-time PCR(qPCR)and Western blotting.Results·The results of CCK-8 showed that the proliferation level of cells in the 10 s,15 s,20 s,and 30 s groups was enhanced from 3 d after irradiation(all P<0.05).After induction of osteogenic differentiation,ALP content,activity,and calcium salt level in the laser irradiation groups increased with the extension of irradiation time(all P<0.05).The results of qPCR and Western blotting analysis showed that the expression levels of the DKK-1 gene and protein in the laser irradiation groups decreased with the extension of irradiation time.However,the expression levels of β-catenin and RUNX2 genes and proteins increased significantly with the extension of irradiation time;there were statistically significant differences between the 15 s,20 s,and 30 s groups and the control group(all P<0.05).Conclusion·Nd:YAP laser biostimulation may promote proliferation and osteogenic differentiation of hPDLCs through WNT/β-catenin signaling pathway.

The intracellular parasite Babesia microti is among the most significant species causing human babesiosis and is an emerging threat to human health worldwide. Unravelling the pathogenic molecular mechanisms of babesiosis is crucial in developing new diagnostic and preventive methods. This study assessed how priming with B. microti surface antigen 1 (BHSA 1) and seroreactive antigen 5-1-1 (BHSA 5-1-1) mediate protection against B. microti infection. The results showed that 500 µg/ml rBMSA1 and rBMSA5-1-1 partially inhibited the invasion of B. microti in vitro by 42.0 ± 3.0%, and 48.0 ± 2.1%, respectively. Blood smears revealed that peak infection at 7 days post-infection (dpi) was 19.6%, 24.7%, and 46.7% in the rBMSA1, rBmSA5-1-1, compared to the control groups (healthy mice infected with B. microti only), respectively. Routine blood tests showed higher white blood cell, red blood cell counts, and haemoglobin levels in the 2 groups (BMSA1 and BMSA5 5-1-1) than in the infection control group at 0–28 dpi. Moreover, the 2 groups had higher serum interferon-γ, tumor necrosis factor-α and Interleukin-17A levels, and lower IL-10 levels than the infection control group throughout the study. These 2 potential vaccine candidate proteins partially inhibit in vitro and in vivo B. microti infection and enhance host immunological response against B. microti infection.

Objective: To investigate the effect of skin soft tissue expansion on repair of large area of scars on extremities. Methods: Twenty-five patients with large area of scars on extremities were admitted to our department from June 2007 to October 2014. There were 14 males and 11 females, aged 4 to 36 years. Operations were performed under local infiltration anesthesia or general anesthesia. In the first stage, 1 to 5 cylindrical expanders with capacities of 250 to 600 mL were placed at left or right sides or at upper or lower parts of the scars. In the second stage, scars of 21 patients were repaired with expanded transverse propulsive and lateral flaps, and scars of 4 patients were repaired with expanded perforator flaps whose pedicles were perforators of brachial artery, superior ulnar collateral artery, or posterior interosseous artery according to areas and shapes of the scars. The secondary wound areas ranged from 13 cm×7 cm to 34 cm×18 cm after dissolution or excision of scars. The areas of flaps ranged from 13 cm×7 cm to 20 cm×12 cm. The donor sites were sutured directly. The flaps after operation and follow-up of patients were observed and recorded. Results: All expanded flaps survived after operation. And the superficial distal part of flap whose pedicle was perforator of posterior interosseous artery in one patient was with necrosis, and other flaps survived well. During follow-up of 3 to 15 months after operation of the second stage, color and texture of flaps were similar to surrounding skin, while extremities of donor sites were thinner and auxiliary incisional scars formed after expansion. Conclusions Expanded flap is a good way to repair large area of scar on extremities. Bilateral skin of scar is the first choice of donor site of expanded flap. If there isn′t enough skin for expanding on bilateral sides, expanded perforator flap designed at upper or lower part of the scar is another choice to repair the scar.

Posterior tibial slope (PTS) is used to describe the sagittal alignment of the tibial plateau of the knee. As its values indicate the steepness or gentleness of the tibial platform, it is an important basis for knee surgery, such as total knee replacement (TKA), anterior cruciate ligament/posterior cruciate ligament reconstruction (RACL/RPCL) and tibia high-level osteotomy (HTO), and affects the indication and efficacy of knee surgery. Since there has been no consensus description of PTS at present in clinical practice in China, this paper intends to discuss PTS from perspectives of epidemiology, measurement, its influence on knee joint activity, relationship between subchondral bone and knee ligament, and its significance in various knee joint operations. This review hopes to contribute to the knee surgery after the surgeons have a comprehensive understanding of the clinical significance and applications of PTS.

Objective: To investigate the therapeuctic effect of one-stage reconstruction of distal urethra with free graft of tublar oral mucosa. Methods: Two strips of oral mucosa graft( 0.4—0.6 cm in width), were harvested and sutured around an oiled silk roll to form mucosa tube. The mucosa tube was used to reconstruct distal urethra. Postoperative pressure dressing and earlier urination were recommended. Results: From May 2007 to October 2015, 16 cases with distal urethra defect or stenosis were treated with this method. The urethra defect was 2—4 cm in length. Urethral fistula happened in 3 patients. All the other 13 cases healed primarily. 10 cases were followed up for 1—5 years by telephone with normal function. Conclusions One-stage reconstruction with free graft of bulbar oral mucosa is suitable and reliable for distal urethra defect less than 4 cm in length.

Objective To optimize the formulation of the sunscreen used in military training.Methods The different formulations of sunscreen were prepared according to the orthogonal design,and then applied on 3M adhesive tape posted on the color dish with a certain amount;the absorbance in 280 nm-320 nm and 320 nm-400 nm of them were measured and the arithmetic mean (A) was concluded.By comparing A,the protective effect and the main factors affecting preventive bask were determined.The preventive bask test of the optimum formula was carried out in human trials using the method of multi point increasing ultraviolet irradiation.Results OMC,TinosorbTM M,rutin,pearl powder and baicalin all had significant effects in preventing UVA.OMC had a significant effect in preventing UVB.The formula 1 (containing the five materials mentioned above) had the best effects in preventing both UVA (SPF 18) and UVB (PA+).Conclusions The formula 1 has the best effects in preventing both UVA and UVB.