[Expression and distribution of calprotectin in healthy and inflamed periodontal tissues].

OBJECTIVE: Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect. METHODS: Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry. RESULTS: Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining. CONCLUSION: Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

[Preclinical study of the effects of fluorinated porcine hydroxyapatite in repairing peri-implant bone defects in canine mandible].

Objective: To evaluate the effects of fluorinated porcine hydroxyapatite (FPHA) on guided bone regeneration of peri-implant buccal bone defects in canine mandible. Methods: Six male beagle dogs were randomly divided into two groups with different time points (4 weeks and 12 weeks after implants placement), with 3 dogs in each group. Bilateral mandibular second premolars, first molars, and second molars in each dog were extracted. The wounds were allowed to heal for 12 weeks. For each dog, four implant beds were prepared in each side and standardized peri-implant buccal bone defect was created at each implant site. After implants placement, the defect sites were randomly allocated in a split-mouth design to blank control group, deproteinized bovine bone mineral (DBBM), the porcine hydroxyapatite (PHA), FPHA and covered with collagen membranes. The animals were sacrificed 4 or 12 weeks after the surgery. Biopsies of the implant sites were obtained for micro-CT evaluation [bone volume fraction (BV/TV) and bone trabecular separation degree (Tb.Sp)] and histological analysis. Results: Micro-CT results showed that 4 weeks after implants placement, PHA, FPHA and DBBM successfully maintained the contour of alveolar ridge at the buccal aspect of the implants, while the contour of alveolar ridge collapsed in the blank control group. BV/TV in the FPHA group [(24.77±2.20) %] was significantly higher than that in the PHA group [(16.89±1.70)%] and DBBM group [(15.68±3.15)%] (P<0.05). Tb.Sp in the FPHA group (0.70±0.07) was significantly lower than that in the DBBM group (1.03±0.19) (P<0.05). Twelve weeks after implants placement, the alveolar ridge contour of the grafted sites in PHA, FPHA and DBBM group remained stable. The alveolar ridge of the blank control group was still collapsed. There was no significant difference in BV/TV and Tb.Sp between PHA group, FPHA group and DBBM group. The histomorphological analysis showed that 4 weeks after implants placement, in the central area of the defect, the amount and maturity of new bone (NB) around the material particles in FPHA group was higher than that in PHA group and DBBM group. Osseointegration could be observed between the NB and implant surface in all the four groups. Twelve weeks after implants placement, the material particles were surrounded by a large number of mature NB in PHA, FPHA and DBBM group. Conclusions: The incorporation of fluoride ion into PHA could effectively promote the repair of peri-implant bone defects in the early stage of guided bone regeneration.

[Preliminary study on the expression and distribution of S100A8 and S100A9 in healthy and experimental periodontitis tissues].

Objective: To investigate the systemic expression profile of S100A8 and S100A9 in healthy and inflamed periodontal tissues. Methods: Experimental periodontitis models were established by ligations around the mandibular second molars of six beagle dogs for 12 weeks (ligation group). The mandibular second molars on the opposite side were kept clean (healthy control group). The expressions of S100A8 and S100A9 in healthy and inflamed periodontal tissues of six beagle dogs were examined by immunohistochemistry. The expressions of S100A8 and S100A9 in primary human gingival fibroblasts (hGF) from 3 subjects and human periodontal ligament cells (hPDLC) from 3 other subjects were detected by immunocytochemistry. Results: After the ligation for 12 weeks, the mean probing depth of ligation group [(3.86±0.14) mm] was significantly higher than that of healthy control group [(2.11±0.28) mm] (P<0.01). Results of immunohistochemistry analysis indicated that S100A8 and S100A9 could be expressed in gingival epithelial cells and might infiltrated neutrophils in the healthy periodontium. Except for the gingival epithelial cells and neutrophils, both proteins were induced and expressed in gingival fibroblasts, periodontal ligament cells, microvascular endothelial cells and bone marrow fibroblasts under inflammatory conditions. The distribution of S100A8 and S100A9 differed in the healthy oral gingival epithelium (OGE), which becomes consistent in inflamed OGE. Additionally, the expressions of S100A8 and S100A9 were confirmed in primary hGF and hPDLC. Conclusions: Periodontal inflammation might enlarge the expression scope of S100A8 and S100A9 and enrich multiple cells with expressions of S100A8 and S100A9.

[Pregnancy with new coronavirus infection: clinical characteristics and placental pathological analysis of three cases].

Objective: To investigate the clinical characteristics and placental pathology of 2019-nCoV infection in pregnancy,and to evaluate intrauterine vertical transmission potential of 2019-nCoV infection. Methods: The placentas delivered from pregnant women with confirmed 2019-nCoV infection which were received in the Department of Pathology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology by February 4, 2020 were retrospectively studied. Their clinical material including placental tissue and lung CT, and laboratory results were collected, meanwhile, nucleic acid detection of 2019-nCoV of the placentas were performed by RT-PCR. Results: Three placentas delivered from pregnant women with confirmed 2019-nCoV infection, who were all in their third trimester with emergency caesarean section. All of the three patients presented with fever (one before caesarean and two in postpartum), and had no significant leukopenia and lymphopenia. Neonatal throat swabs from three newborns were tested for 2019-nCoV, and all samples were negative for the nucleic acid of 2019-nCoV. One premature infant was transferred to Department of Neonatology due to low birth weight. By the end of February 25, 2020, none of the three patients developed severe 2019-nCoV pneumonia or died(two patients had been cured and discharged, while another one had been transferred to a square cabin hospital for isolation treatment). There were various degrees of fibrin deposition inside and around the villi with local syncytial nodule increases in all three placentas. One case of placenta showed the concomitant morphology of chorionic hemangioma and another one with massive placental infarction. No pathological change of villitis and chorioamnionitis was observed in our observation of three cases. All samples from three placentas were negative for the nucleic acid of 2019-nCoV. Conclusions: The clinical characteristics of pregnant women with 2019-nCoV infection in late pregnancy are similar to those of non-pregnant patients, and no severe adverse pregnancy outcome is found in the 3 cases of our observation. Pathological study suggests that there are no morphological changes related to infection in the three placentas. Currently no evidence for intrauterine vertical transmission of 2019-nCoV is found in the three women infected by 2019-nCoV in their late pregnancy.

[Analysis of clinical characteristics and factors associated with short term outcomes in early term neonates].

Objective: To investigate the clinical characteristics of early term and full term neonates, and analyze the risk factors associated with short term outcomes in early term neonates. Method: Neonates with birth weight (BW) ≥2 500 g from year 2013 were analyzed retrospectively based on American Congress of Obstericians & Gynecologists (ACOG) latest definition of term infants. According to inclusion and exclusion criteria, early term (gestational age 37-38 weeks) and full term(gestational age 39-40 weeks) neonates were included, whose morbidity constituent proportion was analyzed by χ(2) test or Fisher accuracy test or t test or Wilcoxon test. Risk factors associated with short term outcomes in early term population were analyzed by Logistic regression analysis. Result: There were 3 002 discharged term infants being investigated, among whom 1 303 cases were included(768 males and 535 females), and 37, 38, 39 and 40 weeks' gestational age newborns were 160, 324, 450 and 369 respectively. Compared with full term neonates(n=819), early term neonates (n=484) had longer length of hospital stay (LOS)(6.0(5.0, 9.0) vs. 6.0(4.0, 8.0), Z=2.830, P=0.005), higher usage rate of intravenous antibiotics(86.4%(418/484) vs. 80.1%(656/819), χ(2)=8.009, P=0.005), higher assisted ventilation rate(9.5%(46/484) vs. 2.9%(24/819), χ(2)=25.528, P<0.01), higher pulmonary surfactant administration rate(4.3%(21/484) vs. 1.1%(9/819), χ(2)=14.006, P<0.01), as well as higher hypoglycemia incidence(3.9%(19/484) vs. 1.2%(10/819), χ(2)=10.226, P=0.001). There were no statistically significant differences in 1 min Apgar score (9(9, 10)vs. 9(9, 10), Z=0.860, P=0.390), 5 min Apgar score (10(9, 10) vs. 10(9, 10), Z=0.810, P=0.418), white blood cell count (15 (11, 21) ×10(9) /L vs.15 (11, 22) ×10(9) /L, Z=0.880, P=0.379), hemoglobin count(180 (159, 205) vs. 182 (160, 204) g/L, Z=0.560, P=0.576), or platelet count(303(234, 372) ×10(9)/L vs. 301(237, 391) ×10(9)/L, Z=0.550, P=0.584). BW between 2 500 g and 2 999 g(OR 1.69, 95% CI: 1.10-2.62, χ(2) =5.614, P=0.018), wet lung(OR=2.61, 95% CI: 1.61-4.24, χ(2)=15.023, P=0.000)and pneumonia(OR 1.88, 95% CI: 1.14-3.08, χ(2)=6.192, P=0.013) were risk factors in early term neonates' short term adverse outcomes. Conclusion: Early term newborns are still at their "immature" state, and respiratory disorders are major risk factors associated with short term outcomes. Hence, early delivery during 37-38 weeks should be avoided as possible as we can.

[Comparison of human papilloma virus infection status between men who have sex with men recruited from gay bathhouses and HIV voluntary counseling and testing clinics respectively in Urumqi].

Objective: To understand the human papilloma virus (HPV) infection status in men who have sex with men (MSM) recruited from gay bathhouses and HIV voluntary counseling and testing (VCT) clinics in Urumqi, Xinjiang Uygur autonomous region, and identify the associated risk factors. Methods: A total of 200 MSM aged ≥18 years were recruited by using the " snowballing" sampling method from gay bathhouses and VCT clinics in Urumqi during March-May, 2016. The MSM recruited completed questionnaires after filling in the informed consent form. The information about their demographic characteristics and sexual behaviors were collected, and anal swabs were collected from them for HPV genotyping. Results: The overall HPV infection rate was 54.0%. The HPV infection rate was 66.7%(74/111) in MSM from gay bathhouses and 38.2%(34/89) in MSM from VCT clinics and the high risk type HPV infection rate was 39.6% (44/111) in MSM from gay bathhouses and 14.6% (13/89) in MSM from VCT clinics, the differences were significant (χ(2)=16.112, P<0.05; χ(2)=15.190, P<0.05). Multiple logistic regression analysis indicated that the major risk factors for HPV infection included activity in gay bathhouse (OR=3.732, 95% CI: 1.950-7.141) and anal sexual behavior (OR=2.555, 95%CI: 1.329-4.912). Conclusion: The prevalence of HPV in MSM from gay bathhouses was higher than that in MSM from VCT clinics, indicating that close attention should be paid to the behavior intervention in MSM.

[HIV infection and associated factors in men who have sex with men and also with women in Urumqi from 2010 to 2014].

Objective: To understand the demographic characteristics, sexual behaviors and the factors associated with HIV infection in men who have sex with men and also with women (MSMW) in Urumqi. Methods: According to the "National AIDS Sentinel Surveillance Protocol" , the data of MSM's demographic characteristics, sex behavior histories and serological detection results were collected through Urumqi AIDS sentinel surveillance system during 2010-2014. In MSM, those also had heterosexual behaviors were considered as MSMW. Results: Of 2 073 MSM surveyed, 216 (10.4%) were MSMW, and 115 MSMW (53.2%) were aged 30>years. Compared with MSM, MSMW were more prone to have commercial homosexual behaviors (OR=2.09, 95%CI: 1.35-3.21), and the condom use rate at the latest homosex was low among them (OR=0.58, 95%CI: 0.40-0.85). The HIV infection rates in MSMW and MSM were 7.4% and 9.0%, respectively, the difference had no significance (χ2=0.604, P>0.05). Multivariate logistic analysis showed that among the MSMW the condom use at the latest homosex (OR=0.24,95% CI: 0.09-0.71), and the number of anal sex in the past week (OR=6.63, 95% CI: 1.47-29.90) were the factors associated with HIV infection, and in MSM, the factors associated with HIV infection were non-Han nationality (OR=6.40, 95% CI:1.62-25.36), local residence time (OR=7.47, 95% CI: 1.98-28.20), using no condoms or using condoms occasionally at commercial sex (OR=6.14,95% CI:1.93-19.52). Conclusion: The HIV infection status and associated factors varied between MSMW and MSM in Urumqi. Therefore, it is necessary to take targeted AIDS prevention measures among them.

ß-Hydroxybutyric acid inhibits growth hormone-releasing hormone synthesis and secretion through the GPR109A/extracellular signal-regulated 1/2 signalling pathway in the hypothalamus.

ß-Hydroxybutyric acid (BHBA) has recently been shown to regulate hormone synthesis and secretion in the hypothalamus. However, little is known about the effects of BHBA-mediated hormone regulation or the detailed mechanisms by which BHBA regulates growth hormone-releasing hormone (GHRH) synthesis and secretion. In the present study, we examined the expression of the BHBA receptor GPR109A in primary hypothalamic cell cultures. We hypothesised that BHBA regulates GHRH via GPR109A and its downstream signals. Initial in vivo studies conducted in rats demonstrated that GHRH mRNA expression in the hypothalamus was strongly inversely correlated with BHBA levels in the cerebrospinal fluid during postnatal development (r = -0.89, P < 0.01). Furthermore, i.c.v. administration of BHBA acutely decreased GHRH mRNA expression in rats. Further in vitro studies revealed a decrease in GHRH synthesis and secretion in primary hypothalamic cells after treatment with BHBA; this effect was inhibited when hypothalamic cells were pretreated with pertussis toxin (PTX). BHBA had no effect on GHRH synthesis and secretion in GT1-7 cells, which do not exhibit cell surface expression of GPR109A. Furthermore, BHBA acutely decreased the transcription of the homeobox gene for Gsh-1 in the hypothalamus in both in vivo and in vitro, and this effect was also inhibited by PTX in vitro. In primary hypothalamic cells, BHBA activated the extracellular signal-regulated kinase (ERK)1/2, p38 and c-Jun N-terminal kinase mitogen-activated protein kinase (MAPK) kinases, as shown by western blot analysis. Moreover, inhibition of ERK1/2 with U0126 attenuated the BHBA-mediated reduction in Gsh-1 expression and GHRH synthesis and secretion. These results strongly suggest that BHBA directly regulates GHRH synthesis and secretion via the GPR109A/ERK1/2 MAPK pathway, and also that Gsh-1 is essential for this function.

Analysis of residual solvents in 2-[(18)F]FDG by GC.

A gas chromatography method has been developed for the measurement of the residual acetone, ethanol and acetonitrile in 2-deoxy-2-[(18)F] fluoro-D-glucose (2-[(18)F]FDG), in accordance with the pending FDA revision on the drug. The detections limits were 0.1 ppm for all three solvents. Good precision and linearity were obtained over ranges spanning the allowable concentration levels proposed by FDA. The amounts of the three solvents in our routine 2-[(18)F]FDG products have been found well below the maximum permissible levels. The method is very amenable to quality control testing for the radiopharmaceutical.

Hair chromium levels in patients with vascular diseases.

The chromium levels in the hair of patients with hyperlipemia and coronary heart disease were found to be similar to those of healthy controls (p greater than 0.2). In patients with cerebral hemorrhage and cerebral thrombosis, significantly higher hair chromium values were observed than in healthy subjects (p less than 0.001). The possible significance of these findings is discussed.