[MiR-132-3p negatively regulates CAMTA1 to promote Schwann cell proliferation and migration and alleviates I-125 seeds-induced exacerbation of facial nerve injury in rats].

OBJECTIVE: To investigate the regulatory effect of miR-132-3p on calmodulin-binding transcription activator 1 (CAMTA1) and Schwann cell activity in rats with facial nerve injury (FNI) treated with I-125 seeds. METHODS: Rat Schwann cells were irradiated with I-125 seeds and transfected with miR-132-3p mimic, miR-132-3p inhibitor or sh-CAMTA1. The expressions of S100B and ß-tubulin Ⅲ in the cells were detected with immunofluorescence assay, and the expressions of miR-132-3p and CAMTA1 protein were determined using RT-qPCR and Western blotting, respectively. EdU staining and Transwell assay were used to evaluate the changes in cell proliferation and migration ability. In a rat model of FNI, I-125 seeds were implanted into the facial tissues near the facial nerve 2 weeks before modeling, and miR-132-3p mimic was injected subcutaneously in the face after modeling. The pathologies of the facial nerve was assessed by HE, LFB and immunofluorescence staining. The targeting relationship between miR-132-3p and CAMTA1 was verified using StarBase v2.0 database and dual-luciferase reporter assay. RESULTS: Rat Schwann cells showed high expressions of S100B and ß-tubulin Ⅲ. I-125 seeds radiation significantly decreased miR-132-3p expression and repressed proliferation and migration of the cells (P < 0.001). Overexpression of miR-132-3p or CAMTA1 knockdown obviously enhanced proliferation and migration of the Schwann cells, while miR-132-3p knockdown produced the opposite effect. MiR-132-3p negatively regulated CAMTA1 expression. In the rat models of FNI, miR-132-3p injection significantly inhibited CAMTA1 expression and attenuated I-125 seeds-induced exacerbation of FNI. CONCLUSION: Overexpression of miR-132-3p suppresses CAMTA1 expression and promotes Schwann cell proliferation and migration to alleviate I-125 seeds-induced exacerbation of FNI in rats.

[Comparison of clinical effects between periodontal endoscopy aiding subgingival debridement and flap surgery].

OBJECTIVE: To compare the clinical effects of periodontal endoscopy aiding subgingival debridement with periodontal flap surgery on residual deep pockets of chronic periodontitis. METHODS: In the single-blind randomized control clinical study, residual deep pockets of periodontitis patients were still existing when re-evaluation after receiving initial periodontal treatment.The pockets which were necessary for bone surgery or guided tissue regeneration surgery were excluded.The sites were divided into test group and control group randomly.Test group sites underwent periodontal endoscopy aiding debridement and control group sites had periodontal flap surgery.At the baseline and 3 months later, parameters, such as plaque index (PLI), bleeding index (BI), probing depth (PD), attachment loss (AL) were examined.Bone height was measured by parallelly digital X-ray dental film.Meanwhile, treatment time and comfort scale (visual analogue scale, VAS) were also recorded. RESULTS: At baseline, 41 proximal sites with residual deep pockets were enrolled (test sites=21, control sites=20).All the parameters were not significantly different between the two groups.PD decreased by (1.67±0.91) mm from (5.62±0.86) mm to (3.95±0.74) mm in test group and by (2.25±1.12) mm from (5.95±1.19) mm to (3.70±0.73) mm in control group significantly (P < 0.05).The difference between the two groups was not significant.The PD of all the sites decreased lower than 5 mm, meanwhile, 76% was lower than 4 mm in test group and 85% was lower than 4 mm in control group.The BI decreased by 0.81±0.93 in test group and 0.65±0.99 in control group significantly (P < 0.05).The difference between the two groups was not significant.The PLI showed a tendency of decrease more in test group and bone height showed a trend of decrease more in control group, however, the difference was not significant.The treatment time was almost the same.The patients felt more comfortable in the test group (VAS was 0.60±0.89) than the control group (VAS was 1.20±1.64), however, the difference was not significant. CONCLUSION: Periodontal endoscopy aiding subgingival debridement could improve periodontal status by reducing the PD and BI significantly in short term.The effect was almost the same with periodontal surgery and endoscopy treatment may decrease the necessity of surgery.Meanwhile, periodontal endoscopy has more comfortable treatment experience than flap surgery and does not need extra treatment time.

Association between Intrinsic Capacity and Sarcopenia in Hospitalized Older Patients.

OBJECTIVES: This study aimed to clarify the association between intrinsic capacity (IC) and sarcopenia in hospitalized older patients. DESIGN: A cross-sectional study. SETTING: Hospital-based. PARTICIPANTS: This study included 381 inpatients aged ≥ 60 years (225 men and 156 women). MEASUREMENTS: IC was evaluated in five domains defined by the World Health Organization: cognition (Mini-Mental State Examination), locomotion (Short Physical Performance Battery test), vitality (Short-Form Mini Nutritional Assessment), sensory (self-reported hearing and vision) and psychological (5-item Geriatric Depression Scale) capacities. IC composite score (0-5) was calculated based on five domains, with lower scores representing greater IC. Sarcopenia was defined in accordance with the criteria recommended by the Asian Working Group for Sarcopenia (AWGS) 2019. Multiple linear and logistic regressions were performed to explore the associations between IC composite score and IC domains with sarcopenia and its defining components. RESULTS: The mean age of 381 patients included was 81.95±8.42 years. Of them, 128 (33.6%) patients had sarcopenia. The median IC composite score was 1 (1, 2). Cognition, locomotion, vitality, sensory and psychological capacities were impaired in 22.6%, 63.5%, 18.9%, 27.3% and 11.3% of patients. Multiple linear regression analyses showed that favorable IC domain scores in cognition, locomotion and vitality were associated with a stronger handgrip strength. A higher vitality score was associated with a greater appendicular skeletal muscle mass index (ASMI), and a higher locomotion score was associated with a greater gait speed. The multiple logistic regression analysis showed that only vitality impairment was associated with sarcopenia. A higher IC composite score was associated with higher risks of sarcopenia, as well as low ASMI, handgrip strength and gait speed. CONCLUSION: This study indicated that a more serious impairment of IC was associated with a greater risk of sarcopenia. Vitality was the domain most strongly associated with sarcopenia. IC may be employed to detect and manage sarcopenia.

[Reverse partial pulmonary resection: a new surgical approach for pediatric pulmonary cysts].

OBJECTIVE: To evaluate the safety and efficacy of reverse partial lung resection for treatment of pediatric pulmonary cysts combined with lung abscesses or thoracic abscess. METHODS: We retrospectively analyzed the clinical data of children undergoing reverse partial lung resection for complex pulmonary cysts in our hospital between June, 2020 and June, 2021.During the surgery, the patients lay in a lateral position, and a 3-5 cm intercostal incision was made at the center of the lesion, through which the pleura was incised and the fluid or necrotic tissues were removed.The anesthesiologist was instructed to aspirate the sputum in the trachea to prevent entry of the necrotic tissues in the trachea.The cystic lung tissue was separated till reaching normal lung tissue on the hilar side.The proximal end of the striated tissue in the lesion was first double ligated with No.4 silk thread, the distal end was disconnected, and the proximal end was reinforced with continuous sutures with 4-0 Prolene thread.The compromised lung tissues were separated, and the thoracic cavity was thoroughly flushed followed by pulmonary inflation, air leakage management and incision suture. RESULTS: Sixteen children aged from 3 day to 2 years underwent the surgery, including 3 with simple pulmonary cysts, 11 with pulmonary cysts combined with pulmonary or thoracic abscess, 1 with pulmonary cysts combined with tension pneumothorax and left upper lung bronchial defect, and 1 with pulmonary herpes combined with brain tissue heterotaxy.All the operations were completed smoothly, with a mean operation time of 129 min, an mean hospital stay of 11 days, and a mean drainage removal time of 7 days.All the children recovered well after the operation, and 11 of them had mild air leakage.None of the children had serious complications or residual lesions or experienced recurrence of infection after the operation. CONCLUSION: Reverse partial lung resection is safe and less invasive for treatment of complex pediatric pulmonary cysts complicated by infections.

[Exploration and practice of Peking University Model to improve the clinical teaching effect of clinical postgraduates majoring in periodontology].

The number of patients with periodontal disease in China is large, and the ratio of doctors to patients is seriously imbalanced, especially in the insufficient number of periodontal specialists and periodontal teachers. Strengthening the cultivation of professional postgraduates in periodontology can effectively solve this problem. This paper summarizes the experiences of Peking University School and Hospital of Stomatology in the teaching of periodontal postgraduate students for more than 30 years, in cluding teaching objectives formulation, teaching resources allocation and enhancement of the quality control system of clinical teaching, for ensuring that the periodontal professional postgraduates could reach the expected level after training. This formed the current "Peking University Model". There are both opportunities and challenges in clinical teaching of periodontal postgraduates in domestic stomatology community. The authors hope that the continuous exploration and improvement of this teaching system will promote the vigorous development of clinical teaching for the postgraduates majoring in periodontology in China.

[Multivariable analysis of tooth loss in subjects with severe periodontitis over 4-year natural progression].

OBJECTIVE: To evaluate the characteristics of severe periodontitis with various number of tooth loss during 4-year natural progression, and to analyze the factors related to higher rate of tooth loss. METHODS: A total of 217 patients aged 15 to 44 years with severe periodontitis were included, who participated in a 4-year natural progression research. Data obtained from questionnaire survey, clinical examination and radiographic measurement. Tooth loss during 4-year natural progression was evaluated. The baseline periodontal disease related and caries related factors were calculated, including number of teeth with bone loss > 50%, number of missing molars, number of teeth with widened periodontal ligament space (WPDL), number of teeth with periapical lesions and etc. Characteristics of populations with various number of tooth loss and the related factors that affected higher rate of tooth loss were analyzed. RESULTS: In 4 years of natural progression, 103 teeth were lost, and annual tooth loss per person was 0.12±0.38. Nine patients lost 3 or more teeth. Thirty-four patients lost 1 or 2 teeth, and 174 patients were absent of tooth loss. Molars were mostly frequent to lose, and canines presented a minimum loss. The number of teeth with WPDL, with periapical lesions, with intrabony defects, with probing depth (PD)≥7 mm, with PD≥5 mm, with clinical attachment loss≥5 mm, with bone loss > 50% and with bone loss > 65% were positively correlated to number of tooth loss. Results from orderly multivariate Logistic regression showd that the number of teeth with bone loss > 50% OR=1.550), baseline number of molars lost (OR=1.774), number of teeth with WPDL (1 to 2: OR=1.415; ≥3: OR=13.105), number of teeth with periapical lesions (1 to 2: OR=4.393; ≥3: OR=9.526) and number of teeth with caries/residual roots (OR=3.028) were significant risk factors related to higher likelihood of tooth loss and multiple tooth loss. CONCLUSION: In 4 years of natural progression, the number of teeth with bone loss > 50%, baseline number of missing molars, number of teeth with WPDL, baseline number of teeth with periapical lesions and number of teeth with caries/residual roots were significantly related to higher risk of tooth loss and multiple tooth loss among Chinese young and middle-aged patients with severe periodontitis in rural areas.

[A survey on human resources of dentists who deal with periodontal disease in Beijing].

Objective: To investigate the status quo of human resources of dentists who deal with periodontal disease in Beijing area through an online survey, which may hopefully provide a preliminary basis for the decision-making of administrative departments and the formulation of periodontal professional development plan. Methods: The dentists who deal with periodontal disease at least half a day per week in Beijing area were investigated. A questionnaire was designed by the chairman of the Periodontology Committee of Beijing Stomatological Association. The questionnaire was sent to and finished by the dentists via "WenJuanXing" online survey software. The contents of the survey included general condition, the property of practice unit, title and position of the dentist, membership of professional society, time and content of periodontal treatment, adoption of new technology and new method of periodontal therapy during the past one year, status of periodontal treatment in the local population and reasons, understanding and influencing factors of periodontal professional development. Results: A total of 1 255 dentists completed the survey, who came from all 16 districts in Beijing, mainly Haidian, Chaoyang, Dongcheng and Xicheng Districts [The total percentage of these four main districts was 70.3% (882/1 255)]. The mean age of the dentists was (36.1±8.3) years. Among the dentists, 71.1% (892/1 255) were females, 88.1% (1 106/1 255) got a Bachelor's degree or above. It was estimated that 35.4% (444/1 255) of the dentists had received standardized periodontal training ever. The percentage of dentists carrying out new technology in the past one year was as high as 68.1% (855/1 255). There were only 163 periodontal specialists (13.0%) out of the dentists in the survey. Only 15.9% (200/1 255) of the dentists routinely performed periodontal surgery. The majority of the dentists [82.8% (1 039/1 255)] were from the state-owned hospitals. Fifty-four point seven percemt (686/1 255) of the dentists thought that lack of knowledge was the main reason why the general public failed to receive periodontal treatment. As for the biggest bottleneck affecting periodontal professional development, fifty-one point zero percent (640/1 255) of the dentists attributed it to the public neglect. Conclusions: The periodontal practitioners in Beijing are young, highly educated, unevenly distributed in 16 districts and mostly females. State-owned oral health institutions are an important force in periodontal diagnosis and treatment services in Beijing. The number of periodontal specialists need to be improved. Promotion of standardized periodontal surgery and the popularization of healthcare knowledge on periodontal disease should also be the focus in the future.

[Epidemiological investigation of tinnitus in Sichuan and Chongqing].

Objectives: To investigate the prevalence and associated risk factors of tinnitus in Sichuan and Chongqing. Methods: We designed a tinnitus epidemiological questionnaire. The multi-stage stratified cluster random sampling methods was applied to obtain study subjects in six areas (Nanchong, Jiangjin, Fengdu, Yunyang, Suining and Ya'an), which were selected for epidemiological investigation. Home visit completion of epidemiological questionnaires was conducted. The trained investigators guided the respondents to fill in the tinnitus epidemiological questionnaires, and the epidemiological status of six areas on prevalence and risk factor was investigated. SPSS 22.0 software was used for statistical analysis. Results: Sampling population were 10 289, in which 9 273 were valid questionnaires. There were 4 281 males and 4 992 females, with an average age of 47.3 years, among which 34.83% (3 230/9 273) had tinnitus. 3.99% (370/9 273) were diagnosed with bothersome tinnitus. In a multivariable logistic regression mod, the following factors were associated with onsetting of tinnitus: sleep disorder [Odds Ratio(OR)=3.74] and noise exposure(OR=1.99). The risk of disease was lowest in the age of 30-40 years old, while the risk of disease was higher for people under 30 and over 40. In another multivariable logistic regression mode, the following factors were associated with having bothersome tinnitus: older people were more likely to suffer from tinnitus, sleep disorders (OR=4.68) and noise exposure (OR=1.56). Conclusions: The prevalence of tinnitus in Sichuan and Chongqing is about 34.83%, but most of the tinnitus is short-lived and has low loudness, which will not affect the patients. Only a small number of patients with tinnitus (3.99%) persist and affect their health and need treatment. The occurrence and exacerbation of tinnitus may be related to sleep, age, and noise exposure.

[Gingival thickness assessment of gingival recession teeth].

OBJECTIVE: To evaluate the gingival thickness and gingival biotype of gingival recession teeth of Chinese population. METHODS: A total of 112 non-molar teeth with gingival recession in 34 patients were included. Direct measurement, cone-beam computerized tomography (CBCT) measurement and periodontal probe method were used to evaluate gingival thickness and biotype. Gingival thickness was measured at 2 mm apical to the gingival margin. Direct measurement was performed with a caliper of 0.01 mm resolution and anesthesia needles attached to silicone disk stops. Gingival biotype was assessed by sulcus probing, if the periodontal probe was visible through the gingival tissue, the gingival biotype was thin; If not visible, the gingival biotype was thick. The differences of gingival thickness among different gingival biotype, tooth site and gingival recession type were analyzed respectively. Besides, the results of CBCT measurement was analyzed compared with the direct measurement. RESULTS: The average gingival thickness of non-molar recession teeth was (1.17±0.41) mm. The average gingival thickness of thick and thin biotype group were (1.38±0.4) mm and (0.97±0.30) mm, respectively, with statistically significant difference (P<0.001). The median of gingival thickness was 1.1 mm. Using 1.1 mm as the cut-off value of thick and thin gingival thickness group, the results matched well with the gingival biotype classification results by periodontal probe method (P=1.000). The average gingival thickness of maxillary teeth was significantly thicker than that of the mandibular teeth. They were (1.39±3.44) mm and (1.01±0.31) mm, respectively (P<0.001). The mean gingival thickness of MillerI, II and III degree gingival recession teeth were (1.15±0.34) mm, (0.83±0.17) mm and (1.26±0.56) mm, respectively, without statistically significant difference (P=0.205). The gingival thickness measurement results between CBCT method and direct measurement were without statistically significant difference (P=0.206). CONCLUSION: In the non-molar gingival recession teeth, the cut-off value of gingival thickness to classify thick and thin biotype of Chinese population was 1.1 mm. The average gingival thickness of the maxillary teeth was significantly thicker than that of the mandibular teeth. Besides, CBCT measurement was an accuracy method for evaluating facial gingival thickness.

[Role of growth arrest-specific protein 6 in migration and osteogenic differentiation of human periodontal ligament cells].

OBJECTIVE: To investigate the role of growth arrest-specific protein 6 (Gas6) in the process of the migration and osteogenic differentiation of human periodontal ligament cells (hPDLCs). METHODS: After different concentrations of recombinant human Gas6 (rhGas6) were added to hPDLCs, cell prolife-ration experiment (CCK-8) was taken to observe the effect of rhGas6 on hPDLCs cell proliferation. Scratch test and cell migration test (Transwell) were taken to analyze the migratory ability of hPDLCs in different concentrations of rhGas6 groups. After osteogenic induction, real-time quantitative polymerase chain reaction (real-time PCR) was taken to detect the expression of the Runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP). ALP staining was used to detect the amount of mineralized nodules. RESULTS: After adding different concentrations of rhGas6, there were no statistically significant differences in hPDLCs cell proliferation among the experimental groups and the control group at 24, 48 and 72 hours (P>0.05). After 24 h of scratch, the healing area in the 800 µg/L of the rhGas6 group was greater than that in the control group, but without statistically significant difference (31.06%±13.70% vs. 21.79%±9.51%, P>0.05). In the migration test, after 24 h, the number of hPDLCs cells which penetrated through the membrane in the 800 µg/L rhGas6 group was significantly higher than that in the control group (P < 0.01). After rhGas6 was added and osteogenic induction, Runx2 and ALP gene expressions of hPDLCs in the 800 µg/L group were significantly higher than those in the control group (1.60±0.30 vs. 0.91±0.10, 2.81±0.61 vs. 0.86±0.12, P < 0.01). After Gas6 was knocked down, the ALP expression of hPDLCs was significantly lower than that of the control group (0.39±0.07 vs. 0.92±0.14, P < 0.01). There was no significant change in Runx2 expression (P>0.05). After 7 days of osteogenic induction, the mineralized nodules formed in the Gas6 knockdown group were significantly less than those in control group (0.25±0.04 vs. 1.00±0.11, P < 0.001). After 14 days of induction, the staining degree of the Gas6 knockdown group was lower than that of the control group, but there was no significant difference (0.86±0.04 vs. 1.00±0.16, P>0.05). CONCLUSION: After downregulation of Gas6 gene, mineralized nodule formation was reduced and ALP gene expressions were decreased in the early stage of osteogenic induction (7 days). After addition of rhGas6, Runx2 and ALP gene expressions were increased and the number of cell migration was increased, suggesting that Gas6 might play a promoting role in the migration and osteogenic differentiation of human periodontal ligament cells.

[Comparison of two training protocols in periodontal surgery teaching for undergraduate students].

Objective: To compare the efficacies of two training protocols, i.e. the multimedia instruction and the conventional method, in periodontal surgery teaching for undergraduate students. Methods: One hundred and twenty-three dental undergraduates in their pre-clinical training course were recruited and divided into two groups according to the learning grade matching principle: the experimental group (multimedia instruction, 60 students) and the control group (conventional method, 63 students). The teaching aim was to train the students gingivectomy and periodontal flap surgery by using the pig jaws. The conventional teaching method of teacher-demonstrating and student-practicing was used in the control group, and the practice time of the students' for each surgery was 45 minutes. A standardized teaching video combined with the teachers' explanations of the key steps was used in the experimental group. The students' practice time for each surgery was 60 minutes. The efficacy of teaching protocol was evaluated by the teachers according to the scoring criteria set by the teaching group. Results: In gingivectomy training, accuracy of fixed points, angle of postoperative gingival margin and morphology of gingiva of the experimental group were better than those of the control group. The experimental group also had more complete and continuous excised gingivae and more thorough adjacent gingival removal. The total scores of the experimental group were significantly higher than those of the control group (92.8±2.6 vs. 89.9±3.7, P<0.05). In periodontal flap surgery training, the experimental group operated the blade around the shape of the tooth better in internal bevel incision than the control group. Additionally, the incision position of internal bevel incision, periosteal integrity after flapping and flap depth of the experimental group were better than those of the control group. Besides, the experimental group had smoother flap edge and more thorough debridement. The total scores of the experimental group were significantly higher than those of the control group (92.2±4.1 vs. 89.2±4.4, P<0.05). Conclusions: The teaching efficacy of multimedia instruction was better than that of the conventional method. Its value needs to be further tested in future teaching practice.

[Vestibular incision subperiosteal tunnel access with connective tissue graft for the treatment of Miller classI and II gingival recession].

OBJECTIVE: To evaluate the clinical outcomes of vestibular incision subperiosteal tunnel access (VISTA) with connective tissue graft (CTG) in the treatment of Miller classes I and II localized gingival recession. METHODS: Ten patients with 10 Miller classes I and II localized gingival recessions were enrolled in the study. All defects were equal to or above 2 mm in recession depth. All the patients received treatment with VISTA+CTG. Their clinical parameters, including recession depth (Rec), recession width (RW), keratinized tissue width (KT), clinical attachment loss (CAL), probing depth (PD) were recorded and compared before surgery and 6 months later. The mean root coverage (MRC) and complete root coverage (CRC) were calculated at the end of 6 months. A visual analogue scale (VAS) was used to estimate the patients' discomfort during the operation and during the 2 weeks post-operation. Patient-based aesthetic satisfaction 6 months after surgery was evaluated by a VAS. RESULTS: The mean Rec was (2.65±0.82) mm at baseline, and (0.35±0.58) mm after 6 months. The VISTA+CTG treatment resulted in an improvement of (2.30±0.98) mm in recession depth (P<0.001). MRC was 86.67%±21.94% and CRC reached 70% at the end of 6 months. KT increased (0.90±1.22) mm (P<0.05). Aesthetic satisfaction on the patients' level was 8.30 based on VAS (0=unsatisfied, 10=extremely satisfied). The patients' discomfort during the operation and 2 weeks post operation were 2.40 and 4.30 (0=no pain, 10=extreme pain). Furthermore, clinical outcomes showed no statistically significant difference between the gingival biotypes, and between the teeth positioned in maxillary and in mandibular. CONCLUSION: VISTA+CTG could be an effective treatment for Miller classes I and II localized gingival recession. Clinical outcomes indicated decrease in recession depth and width, and increase in width of keratinized tissue. Patients suffered little pain during the operation and 2 weeks post-operation of healing and accessed good aesthetic satisfaction. VISTA+CTG could be an option for the treatment of Miller classes I and II localized gingival recession.

Correlation between peripheral skeletal muscle functions and the stable phase of COPD in older patients.

OBJECTIVE: To establish normal values for detection indexes of peripheral skeletal muscle dysfunction (quadriceps femoris) of healthy older subjects, and investigate the functional status of the peripheral skeletal muscle of patients with stable phase COPD. PATIENTS AND METHODS: Patients with stable phase COPD and healthy subjects of similar age were included. The assessments of strength and myoelectricity of the quadriceps femoris were recorded. The twitch tension of the quadriceps femoris (TwQ), quadriceps maximum voluntary contraction (QMVC), and endurance time (Tf) were measured. The multiple-parameter malnutrition index (MNI) was used for overall evaluation of the nutritional status of patients. The femoral muscle volume was estimated. All subjects were subjected to a routine pulmonary function test including indexes such as FEV1, FVC, FEV1/FVC (%), and PEF. Enzyme-linked immunoassay (ELISA) was used to measure the levels of myostatin, tumor necrosis factor-α, TNF-like apoptosis-inducing factor (TWEAK), surface active protein D (SPD), C-reactive protein (CRP), interleukin (IL)-1ß, and IL-6. The cell immunohistochemical method was used to detect the expression of nuclear factor Kappa B (NF-κB). RESULTS: There were significant differences in body weight, BMI, femoral muscle volume, and physical activity scores between the two groups (p<0.01). The MNI of patients in the COPD group was significantly higher than that in the control group (p<0.01). The QMVC of 51 male and 16 female patients decreased. All eight tested cytokines increased in the COPD group but there were only significant differences in four cytokines (p<0.05). CONCLUSIONS: Chronic systemic inflammation is a major risk factor of skeletal muscle dysfunction (SMD) in COPD patients. The levels of SPD, myostatin, TWEAK, and TNF-α decreased significantly in COPD patients.

[Clinicopathologic and molecular features of cribriform morular variant of papillary thyroid carcinoma].

Objective: To investigate the clinicopathologic and molecular features of the rare cribriform morular variant of papillary thyroid carcinoma (CMV-PTC). Methods: The clinicopathologic data of 10 patients with CMV-PTC were retrospectively reviewed. Immunohistochemical (IHC) staining was done using LSAB method. DNA sequencing for APC were applied using Sanger method. BRAF V600E mutation was examined using ARMS method. The cytological, morphological, IHC and molecular features were analyzed. Results: All patients were female at an average age of 27 years old. The tumors were mostly located in the right lobe of thyroid. Fine needle aspiration cytology was performed in three patients; two were diagnosed as suspicious for PTC and one as PTC. Nine tumors presented as solitary nodule and two as multiple nodules in both lobes. Infiltration was demonstrated in three cases. The average size was 2.6 cm. The neoplastic cells were arranged in papillary, cribriform, solid and glandular patterns, with rare or without colloid inside the lumen. The number of morula varied, ranging from zero to many. The neoplastic cells were variably enlarged, showing round, oval or spindle shape. Nuclear irregularity was identified as irregular membrane, nuclear grooves or pseudoinclusion, but no typical ground glass feature. Peculiar nuclear clearing could be observed in the morular cells. IHC staining showed the neoplastic cells were negative for thyroglobulin and p63, but positive for TTF1, cytokeratin 19 and estrogen receptor. Diffuse staining with cytokeratin was seen in the neoplastic cells and the morula. Specific cytoplasmic and nuclear staining of ß-catenin was seen in the neoplastic cells but not the morula. Ki-67 proliferation index was 1%-30%. No recurrence or metastasis was observed. One patient was demonstrated to harbor both somatic and germline mutations of the APC gene, who was found to have adenomatous polyposis and her mother died of colonic carcinoma. No BRAF V600E mutation was detected. Conclusions: CMV-PTC is rare and shows atypical cytological and clinicopathological features, and it is easily misdiagnosed.TG, TTF1, ER and ß-catenin are specific IHC markers for CMV-PTC. The morula is negative for cytokeratin 19, in contrast to squamous metaplasia. Although CMV-PTC has indolent clinical behavior, a definite diagnosis is necessary to rule out the possibility of APC gene mutation and related extra-thyroidal neoplasm, such as FAP and Gardner syndrome.

NLRP6 Induces Pyroptosis by Activation of Caspase-1 in Gingival Fibroblasts.

NLRP6, a member of the nucleotide-binding domain, leucine-rich repeat-containing (NLR) innate immune receptor family, has been reported to participate in inflammasome formation. Activation of inflammasome triggers a caspase-1-dependent programming cell death called pyroptosis. However, whether NLRP6 induces pyroptosis has not been investigated. In this study, we showed that NLRP6 overexpression activated caspase-1 and gasdermin-D and then induced pyroptosis of human gingival fibroblasts, resulting in release of proinflammatory mediators interleukin (IL)-1ß and IL-18. Moreover, NLRP6 was highly expressed in gingival tissue of periodontitis compared with healthy controls. Porphyromonas gingivalis, which is a commensal bacterium and has periodontopathic potential, induced pyroptosis of gingival fibroblasts by activation of NLRP6. Together, we, for the first time, identified that NLRP6 could induce pyroptosis of gingival fibroblasts by activation of caspase-1 and may play a role in periodontitis.

[Role of vitamin K-dependent protein Gas6 in the expression of endothelial cell adhesion molecule-1 and chemokines induced by Porphyromonas gingivalis lipopolysaccharide].

OBJECTIVE: Growth-arrest-specific protein 6 (Gas6) is a vitamin K-dependent protein and involved in cell proliferation, survival, adhesion and migration . Also it has been shown to play an important role in the inflammatory response .The aim of present study was to investigate the role of Gas6 in the process of the expression of adhesion molecules and chemokines of human umbilical vein endothelial cells (HUVECs) induced by Porphyromonas gingivalis lipopolysaccharide(P.g-LPS). METHODS: After up-regulation and down-regulation of the expression of Gas6, the vascular endothelial cells were stimulated with 1 mg/L P.g-LPS for 3 h and 24 h. Real-time quantitative polymerase chain reaction(real-time PCR) was taken to detect the expression of the cell adhesion molecules:intercellular adhesion molecule-1 (ICAM-1) and E-selectin, as well as chemokines:interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1). Wound healing assay was taken to observe the migration ability of endothelium cells in different groups. RESULTS: After 3 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in the down-regulation group was not significantly different from that in the control group,while in the up-regulation group the decrease of E-selectin, ICAM-1, IL-8 and MCP-1 was 81%±0%, 47%±3%, 76% ± 3%, 26% ± 6% respectively. After 24 h of P.g-LPS stimulation, the expression of adhesion molecules and chemokine in down-regulation group was significantly higher than that in control group (2.06±0.07, 1.99±0.11, 3.14±0.15, 1.84±0.03 flod), while these molecules in the down-regulation group was significantly lower than in the control group (29%±1%, 62%±3%, 69%±1%, 41%±2%). Differences were statistically significant (P<0.01). Wounding healing assay showed that down-regulation of Gas6 enhanced migration ability of endothelial cells while up-regulation of Gas6 weakened this ability,which was consistent with the trend of real-time PCR result. CONCLUSION: Down-regulation of the Gas6 gene enhanced the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g- LPS stimulating, while up-regulaiton of the Gas6 gene weakened the expression of ICAM-1, E-selectin, IL-8 and MCP-1 in HUVECs after P.g-LPS stimulating,suggesting that Gas6 may play a role in the process of endothelial cell adhesion.

[Method for recording tension changes of small pulmonary artery and vein using the isolated vessel tension measurement system].

Objective: To establish a standardized method for isolated pulmonary artery and vein rings with different diameter, pressure and length, which could provide a more scientific method for in vitro study of pulmonary vessel diseases. Methods: Male SD rats were anesthetized, and the right ventricular systolic pressure were measured. Small pulmonary artery and vein rings with 200-400 µm in diameter and 2 mm in length were prepared by dissecting pulmonary arteries and veins. The pulmonary vessel rings were mounted in the organ bath by 2 stainless steel wires with diameter of 40 µm. Then the internal circumference of the vessel rings was increased gradually with 100 µm per step. At the same time the vascular tension was recorded by the Myograph System and Acknowledgement data acquisition system, and subsequently the passive length-tension exponential curve was made. The initial tension of the rings was set, equilibrated for another 30 min, and then stimulated with 60 mmol/L KCl 3 times, and the best contractile reactivity was achieved. The contractile reactivity of pulmonary artery rings and endothelial integrity were detected by exposure to 1 µmol/L phenylephrine(PE) and 10 µmol/L acetylcholine(Ach), while the contractile reactivity of pulmonary vein rings was detected by exposure to 1 µmol/L U46619 and 10 µmol/L papaverine. Results: The contraction and relaxation effects of the pulmonary artery rings reached 0.39 mg and 92% when they were stimulated by 1 µmol/L PE and 10 µmol/L Ach. The contraction and relaxation effects of pulmonary vein rings were up to 0.13 mg and 84% when they were exposed to 1 µmol/L U46619 and 10 µmol/L papaverine, respectively. Conclusion: Pulmonary artery and vein rings with appropriate basal tension and optimal vasodilator activity were prepared, and a standardized method of tension experiment for isolated pulmonary artery and vein rings established.

Rheumatoid arthritis fibroblast-like synoviocytes co-cultured with PBMC increased peripheral CD4+ CXCR5+ ICOS+ T cell numbers.

'Circulating' T follicular helper cells (Tfh), characterized by their surface phenotypes CD4+ chemokine receptor 5 (CXCR5)+ inducible co-stimulatory molecule (ICOS)+ , have been identified as the CD4+ T cell subset specialized in supporting the activation, expansion and differentiation of B cells. Fibroblast-like synoviocytes (FLS) are critical in promoting inflammation and cartilage destruction in rheumatoid arthritis (RA), and the interaction between FLS and T cells is considered to facilitate FLS activation and T cell recruitment. However, it remains unknown whether RA-FLS co-cultured with activated peripheral blood mononuclear cells (PBMC) has immunoregulatory effects on peripheral Tfh. In the present study, we co-cultured RA-FLS with or without anti-CD3/CD28-stimulated PBMC. The results showed that RA-FLS co-cultured with stimulated PBMC could increase the numbers of CD4+ CXCR5+ ICOS+ T cells of RA PBMC possibly via the production of interleukin (IL)-6, a critical cytokine involved in the differentiation of Tfh cells. We also observed increased reactive oxygen species (ROS) levels in the co-culture system of RA-FLS and PBMC. The percentage of CD4+ CXCR5+ ICOS+ T cells was decreased when ROS production was inhibited by N-acetyl-L-cysteine (NAC), a specific inhibitor which can decrease ROS production. In addition, we showed that the higher levels of tumour necrosis factor (TNF)-α and IL-1ß in the co-culture system and the blocking of TNF receptor 2 (TNF-R2) and IL-1ß receptor (IL-1ßR) both decreased the numbers of CD4+ CXCR5+ ICOS+ T cells. Our study reveals a novel mechanistic insight into how the interaction of RA-FLS and PBMC participates in the RA pathogenesis, and also provides support for the biologicals application for RA.