OBJECTIVES: This study aimed to clarify the effects of Foxp3 silencing on the expression of inflammatory cytokines in human periodontal ligament cells (hPDLFs) in an inflammatory environment and on cell proliferation and invasiveness, as well as to explore the role of Foxp3 gene in the development of periodontitis. METHODS: An small interfering RNA (siRNA) construct specific for Foxp3 was transfected into hPDLFs. Foxp3 silencing efficiency was verified by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, and the siRNA with the optimum silencing effect of Foxp3 gene was screened. Using lipopolysaccharide to simulate an inflammatory environment in vitro, CCK-8 detected the effect of silencing Foxp3 on hPDLFs proliferation under inflammatory conditions. Wound-healing experiments and transwell assays were conducted to detect the effect of silencing Foxp3 on hPDLF migration under inflammatory conditions. The expression of the inflammatory cytokines interleukin (IL)-6 and IL-8 was detected by RT-PCR and Western blotting under inflammatory conditions. RESULTS: After siRNA transfection, RT-PCR and Western blotting analyses showed that the expression of Foxp3 mRNA in the Foxp3-si3 group decreased significantly (t=21.03, P<0.000 1), and the protein expression of Foxp3 also decreased significantly (t=12.8, P<0.001). In the inflammatory environment, Foxp3 gene silencing had no significant effect on hPDLFs proliferation (P>0.05), and Foxp3 gene silencing promoted hPDLFs migration (P<0.05). Moreover, the expression of IL-6 and IL-8 increased (P<0.05). CONCLUSIONS In an inflammatory environment, Foxp3 gene silencing promoted hPDLFs migration but had no significant effect on hPDLFs proliferation. The expression of inflammatory factors expressed in hPDLFs increased after Foxp3 gene silencing, indicating that Foxp3 gene inhibited inflammation in periodontitis.
Humans ; Cell Proliferation/genetics* ; Cells, Cultured ; Cytokines/metabolism* ; Fibroblasts/metabolism* ; Forkhead Transcription Factors/metabolism* ; Gene Silencing ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Periodontal Ligament/metabolism* ; Periodontitis/metabolism* ; RNA, Small Interfering/metabolism* ; Transcription Factors/metabolism*

Cervical cancer is a common malignant tumor of female reproductive system. The treatment of cervical cancer is based on surgery and radiotherapy (or concurrent chemoradiation). Lower extremity lymphedema (LEL) is a frequent complication after cervical cancer treatment, which significantly affects the quality of life of patients. Both pelvic surgery and radiation for cervical cancer can lead to LEL. The risk factors for LEL are complicated and involving characteristics regarding patient (age, comorbidities, lifestyle, etc.), tumor [International Federation of gynecology and Obstetrics (FIGO) stage, lymph node metastasis, etc.], and treatment (number of resected lymph nodes, removal of circumflex iliac nodes, adjuvant therapy, etc.). Comprehensive measures are proposed to prevent cervical cancer patients from LEL, and further investigations in terms of effectiveness are warranted.

In the practical teaching of large-scale medical equipment courses in medical colleges and universities. There are some problems such as less opportunities for students to operate the equipment which is possessed by teaching hospitals, the large-scale medical equipment in teaching hospitals are in a pathogenic environment, and some equipment such as CT, DR have radiation hazards, which lead to difficulties in practical teaching. These problems restrict the cultivation of students' practical ability, cause disconnections between theoretical teaching and practical teaching, and are not conducive to training "new engineering" and "new medical science" talents. So, this paper puts forward an idea which introduces virtual simulation teaching instruments of large-scale medical equipment into the teaching of medical equipment in medical colleges and universities, uses the physical structures, control systems and functional software of virtual simulation teaching instruments to implement the teaching of "integration of theory and practice", and also sets up validation experiments, comprehensive experiments, designing experiments and research explorative experiments in the teaching process. This kind of teaching can not only realize the high integration of theoretical teaching and practical teaching, but also consolidate students' basic theoretical knowledge, improve their abilities to solve complex engineering problems, and cultivate "new engineering" and "new medical" talents in line with the needs of the new era.

Objective: To investigate the function of virD4 gene in Helicobacter pylori clinical strain SBK. Methods: The virD4 gene segment was obtained through T-A cloning method. The prokaryotic expression vector pET-28a(＋)-virD4 was constructed and transformed into E. coli Rosetta for the expression by induction of IPTG. The recombinant proteins were obtained and purified by KCl dyeing with gel cutting method, and identified via SDS-PAGE analysis. The purified recombinant virD4 protein was used to immunize mice to produce polyclonal antibodies. The titer of the polyclonal antibodies was tested by ELISA and the antigenic specificity was identified by western blot. The purified recombinant virD4 proteins were co-cultured with GES-1 cells followed by detecting the expression of inflammatory cytokines secretion and the ability of cell proliferation. Results: The full length of virD4 gene was 1 728 bp. The sequence shared quite high homology with the virD4 gene of isolate Shi470. The recombinant prokaryotic expression plasmid pET-28a(＋)-virD4 was successfully constructed. The recombinant virD4 proteins were obtained by IPTG induction and purified via KCl dyeing method. SDS-PAGE showed that the relative molecular weight of recombinant virD4 protein was 63 000. The purified proteins were used to immunize mice to obtain the anti-virD4 polyclonal antibodies with the titer 512 000. The reaction between anti-virD4 polyclonal antibodies and recombinant virD4 proteins was highly specific. The recombinant virD4 protein induced inflammatory cytokines secretion and promoted GES-1 cell proliferation. Conclusion The virD4 gene was successfully cloned and highly expressed in prokaryotic expression system, and its antibodies were prepared. The recombinant virD4 protein can induce cytokine secretion and cell proliferation.

Objective To investigate the difference between Han nationality residents and the Hani nationality residents in southern Yunnan about the effects of stroke risk factors on cerebral vascular hemodynamic indicators.Methods In the cohort crowds,Hani nationality outpatient patients were selected as the Hani nationality group,the Han nationality outpatient patients with the same gender and similar age were randomly selected as the Han nationality group with the ratio of nearly 1 ∶ 3.The risk factors of stroke and the differences of the hemodynamic indicators were compared between the two groups.Results The dyslipidemia proportion of the Hani nationality group was lower than that of the Han nationality group [48 cases (11.5 ％) vs.257 cases (17.5 ％),x2 =8.867,P =0.003],and the drinking proportion of the Hani nationality group was higher than that of the Han nationality group [79 cases (18.9％) vs.214 cases(14.6％),x2 =4.498,P =0.034],the other risk factors exposure rates had no statistically significant differences between the two groups (all P ＞ 0.05).The ratio of numbers of cerebral vascular hemodynamic indicators below 75 points(optimum truncation points) in the Hani nationality group and the Han nationality group were 30.3％ and 31.9％,respectively,the proportion of the two groups of integral value distribution had statistically significant difference (x2 =12.614,P ＜ 0.01);Vmean (left:t =0.632,P =0.527;right:t =0.642,P =0.532),Vmin (left:t =-0.349,P =0.727;right:t =-0.540,P =0.589),Wv(left:t =0.297,P =0.767;right:t =1.119,P =0.263),Zcv(left:t =0.297,P =0.767;right:t =1.247,P =0.213),Rv(left:t =0.837,P =0.403;right:t =1.222,P =0.222),DR(left:t =0.720,P =0.482;right:t =0.396,P =0.692),Cp(left:t =0.614,P =0.539;right:t =1.486,P =0.138) and other indicators of the two groups had no statistically significant differences.Conclusion The Hani nationality residents'stroke risk factors and CVHI integral distribution in different age groups are different from Han people in southern Yunnan.

Objective: To establish an HPLC determination method for the quantitative indicators in Qubai granules( stilbene gly-cosides, ferulic acid and paeonol). Methods: A dual-wavelength HPLC assay was used with the following conditions: a chromatogra-phy column ODS (150 mm×4. 6 mm, 5 μm) was used, the mobile phase was methanol: 0. 1% phosphoric acid with gradient elution, the column temperature was 35℃, and the injection volume was 10 μl. Results: The linear ranges of the three active constituents were 1.56-156.00 μg·ml-1(r stilbeneglycoside=0.999 8)、1.00~100.00 μg·ml-1(rferulicacid=0.999 8),1.61~161.00 μg·ml-1(rpaeonol=0. 999 7), respectively, and the average recoveries of the three constituents were between 100. 33% and 100. 76% with the RSDS less than 2% (n=6). Conclusion: The method is simple, stable and reliable, which can be used for the quality control of Qubai granules.

Objective To investigate the exposure to stroke risk factors, the proportion of high?risk individuals, and the relationship between risk factor exposure and impaired cerebrovascular hemodynamics in patients at Pu'er People's Hospital. Methods Between April 2014 and June 2017, this study enrolled inpatients and outpatients of the neurology department of Pu'er People's Hospital who underwent cerebrovascular hemodynamic examinations to evaluate stroke risk. A total of 30 103 (12 793 males and 17 310 females) participants aged 22 to 99 (53.3±14.5) years were included. The distribution of exposure rates for stroke risk factors and proportion of high?risk individuals were determined according to age and sex. The relationships between integral scores of cerebrovascular hemodynamics and exposure to risk factors were also analyzed. Results Exposure rates for risk factors of hypertension, dyslipidemia, diabetes, heart disease, stroke family history, cigarette smoking, alcohol drinking, and overweight or obesity in males were 36.6%, 11.4%, 8.8%, 5.9%, 7.0%, 41.6%, 37.8%, and 51.4%, respectively. The exposure rates in females were 31.2%, 11.3%, 7.6%, 5.5%, 6.8%, 0.7%, 1.6%, and 48.8%, respectively. Differences between age groups for all risk factors were significant (P<0.01). Exposure rates for hypertension, diabetes, cigarette smoking, alcohol drinking, and overweight or obesity in males were significantly higher than in females (P<0.01). High?risk males and females accounted for 41.8% and 35.6% (χ2=119.82, P<0.01) and the rates in both groups increased significantly with age (χ2=1 838.2, 2 881.5, P<0.01). Risk factors including age, hypertension, dyslipidemia, diabetes, heart disease, stroke family history, alcohol drinking, and body mass index were independent predictors of increased individual risk of stroke. Conclusions Exposure levels for stroke risk factors and the proportion of individuals at high risk of stroke were relatively high in the hospital population. Cerebrovascular dysfunction is closely related to exposure to common risk factors for stroke.

Objective To study the relationship between serum Treg cells content and superoxide dismutase(SOD) level with the severity and clinical prognosis in the patients with preeclampsia.Methods Forty cases of preeclampsia in our hospital from September 2015 to September 2016 were selected as the research subjects divided into the mild preeclampsia group(25 cases) and severe preeclampsia groups(15 cases) according to the disease severity,and 20 healthy pregnant women served as the control group.The peripheral blood Treg cells and serum SOD level were compared between the two groups.The relationship between Treg cell content and serum SOD level with the disease severity and prognosis in the patients with preeclampsia was investigated.Results The systolic and diastolic blood pressure in the severe preeclampsia group were(145.3±10.6)mm Hg and(102.3±7.8)mm Hg,the occurrence rates of premature delivery,placental abruption and fetal distress were 33.3%,20.0% and 13.3% respectively,BMI before pregnancy was 25.4±1.3,which were significantly higher than those in the mild preeclampsia group and control group,while the mild preeclampsia group was higher than the control group.The Treg cell content and SOD level in the severe preeclampsia group were (2.3±0.7)×106/L and(2.6±0.7)μg/mL,which were significantly lower than those in the mild preeclampsia group and control group and which in the mild preeclampsia group were (3.4±0.6)×106/L and(4.3±0.9)μg/mL,and significantly lower than those in the control group,the differences were statistically significant(P<0.05).The Logistic regression analysis results indicated that BMI index(P=0.018),Treg cell content(P=0.024) and SOD level(P=0.029) were the independent risk factors for poor pregnancy outcome.Conclusion The peripheral blood Treg cells content and serum SOD level are closely related to poor pregnancy outcome in the patients with preeclampsia.

Objective:To optimize the water extraction process of traditional Chinese medicine ( TMC) Qubai granule. Methods:The orthogonal test was used to study four influencing factors including water amount, soaking time, extraction time and extraction times with dry extract yielding rate and the content of ferulic acid as the evaluation indices. Results:The optimum extraction process was as follows:A2 B1 C2 D2 , namely adding 10-fold amount of water, without soaking in advance, extracting twice with 2 h for each time. Con-clusion:The process is simple, stable and reproducible, which provides basis for the industrial production.

Objective To evaluation the method of rapid detection of Methicillin-resistant Stphylococcus aureus (MRSA) ST239 clones with multiplex PCR assay and investigation of the epidemic status of MRSA blood stream infections in Hefei area.Methods Antibiotic susceptibility testing were applied to MRSA isolates from bloodstream infection,rapid screening and confirmation of MRSA ST239 clones by using multiplex PCR,Multilocus Sequence typing (MLST) and Staphyloccoccal Cassette Chromosome mec(SCCmec) typing.Results 51 of 106 clinic isolates Staphylococcus aureus were identified as MRSA,accounting for 48.1％.The resistance rate of MRSA to erythromycin,aminoglycosides and quinolone were significantly higher than Methicillin Sensitive Staphylococcus aureus (MSSA).Both MRSA and MSSA had a high sensitivity to cotrimoxazole,the sensivity rates were 86.3％ and 94.5％,respectively; 47 of 51 isolates of MRSA that detected by MRSA ST239 rapid screening were ST239 clones.Randomly selected 20 positive screening stains were confirmed as MRSA-ST239-SCCmec Ⅲ by MLST and SCCmectyping.Conclusions In Hefei area,nearly half of MRSA bloodstream infections in clinical isolates are MRSA-ST239-SCCmeclⅢ type and serious multidrug-resistance.The rapid detection of ST239 clones by multiplex PCR is a reliable and effective method for large-scale screening in laboratory.