Objective To investigate the effects of thermosensitive moxibustion on psoriasis model rats'skin;To discuss its mechanism based on JAK3/STAT3 signaling pathway.Methods 5%imiquimod cream was used on the skin of rats back and along the route of the Sanjiao meridian of hand-shaoyang for psoriasis modeling.The SD rats were randomly divided into blank group,model group and treaament group.The treatment group received moxibustion therapy at"Waiguan"for 40 minutes each time for 7 consecutive days,and were divided into moxibustion group and thermosensitive moxibustion group according to sensitization state.HE staining was used to observe pathological changes in skin lesions,and ELISA was used to detect serum tumor necrosis factor(TNF)-α,interleukin(IL)-1 and IL-23 levels.Immunohistochemistry was used to detect the positive expressions of p-JAK3 and p-STAT3 in the skin lesion tissue,and RT-qPCR was used to detect the mRNA expressions of JAK3,STAT3,IL-17A and IL-17F in the skin lesion tissue.Results Compared with the normal group,model group rats were with protruding and thickened skin lesions accompanied by extensive infiltration of inflammatory cells,the contents of serum TNF-α,IL-1β and IL-23 significantly increased(P<0.01),and the positive expressions of p-JAK3 and p-STAT3 in the skin lesion tissue significantly increased(P<0.01),the mRNA expressions of JAK3,STAT3,IL-17A and IL-17F significantly increased(P<0.01).Compared with the model group,the thickness of the spinous layer in the skin lesions of rats in the thermosensitive moxibustion group was reduced,and there was no significant infiltration of inflammatory cells.The contents of serum TNF-α,IL-1β and IL-23 significantly decreased(P<0.01),and the positive expressions of p-JAK3 and p-STAT3 in skin lesion tissue significantly decreased(P<0.05,P<0.01),the mRNA expressions of JAK3,STAT3,IL-17A and IL-17F significantly decreased(P<0.05,P<0.01).Compared with the moxibustion group,the serum contents of TNF-α and IL-1β in the thermosensitive moxibustion group were significantly decreased(P<0.05,P<0.01),and the positive expression of p-JAK3 in the skin lesion tissue was significantly decreased(P<0.05).Conclusion Thermosensitive moxibustion can improve the pathological symptoms of psoriasis model rats.Its mechanism may be related to the inhibition of the JAK3/STAT3 signaling pathway,reduction of regulatory T cell-17 differentiation and secretion of pro-inflammatory cytokines.

Objective:To study the clinical characteristics of ureaplasma urealyticum (UU) infection in preterm infants with gestational age <34 weeks.Methods:From January 2017 to December 2021, premature infants with gestational age <34 weeks admitted to neonatal department of our hospital were enrolled in this prospective cohort study. UU-DNA from respiratory tract samples were examined using quantitative fluorescence polymerase chain reaction method. The infants were assigned into UU (+) group and UU (-) group. Perinatal factors and clinical characteristics were compared between the two groups.Results:A total of 182 preterm infants were enrolled, including 59 cases (32.4%) in UU (+) group and 123 (67.6%) in UU (-) group. UU (+) group had significantly lower gestational age and birth weight and significantly higher incidences of vaginal delivery, premature rupture of membranes (PROM) >18 h and maternal chorioamnionitis than UU (-) group ( P<0.05). Compared with UU (-) group, UU (+) group had significantly higher leucocyte count, neutrophil count and interleukin-6 at 1, 24 and 72 h after birth ( P<0.05). No significant differences existed in C-reactive protein and procalcitonin between the two groups at each time point ( P>0.05). In UU (+) group, the incidences of intrauterine pulmonary infection and bronchopulmonary dysplasia (BPD) were higher and the incidence of respiratory distress syndrome was lower than UU (-) group ( P<0.05). No significant differences existed in the incidences of intraventricular hemorrhage, periventricular leukomalacia, feeding intolerance, necrotizing enterocolitis, retinopathy of prematurity between the two groups ( P>0.0 5). UU (+) group had significantly longer duration of oxygen therapy than UU (-) group ( P<0.05). No significant differences existed in the duration of invasive mechanical ventilation and hospital stay between the two groups ( P>0.05). Conclusions:Preterm infants <34 weeks with positive UU in respiratory tract secretions have higher incidences of vaginal delivery, PROM>18 h and maternal chorioamnionitis. Leukocyte and neutrophil count and interleukin -6 are higher in these infants. They need prolonged oxygen therapy and have increased risks of intrauterine pulmonary infection and BPD.

Objective　To explore the predictive value of the ratio of C-reactive protein to high-density lipoprotein cholesterol (CHR) for carotid plaque vulnerability in patients with ischemic stroke.Methods　A retrospectively enrolled 349 patients with acute ischemic stroke in the Department of Neurology of Baoding First Central Hospital from January 2019 to January 2020.All subjects underwent ultrasound examination to evaluate the condition of carotid artery plaques,and based on the ultrasound results,they were divided into no plaque group,stable plaque group,unstable plaque group.The differences in risk factors,blood lipids,and inflammatory indicators in each group were compared.The receiver operating characteristic (ROC) curve was used to estimate the predictive value of each index on the vulnerability of carotid plaque.Results　C-reactive protein and CHR of the unstable plaque group were higher than those of the stable plaque group,and the difference was statistically significant (P<0.05).ROC curve shows that CHR can predict carotid plaque instability more reliably than a single C-reactive protein.Conclusion　CHR has high predictive value for carotid artery vulnerable plaque and can be used as a new circulating biomarker to identify carotid artery plaque vulnerability.

Objective: To investigate the value of short tandem repeat (STR) genotyping in the diagnostic workup of molar and non-molar gestations with correlation of histological characteristics. Methods: Six hundred and fifty-six cases were selected based on clinically suspected hydropic abortion and/or molar pregnancy from July 2015 to September 2017 at Beijing Obstetrics and Gynecology Hospital. DNA was extracted from dissected chorionic villi and paired maternal endometrial FFPE tissue samples by Simplex OUP™ FFPE DNA Tissue Kit. STR genotyping was performed by PowerPlex 16 HS system. Results: DNA genotyping was informative in 649 of 656 cases, leading to identification of 215 hydatidiform mole gestations and 434 non-molar gestations. Most of non-molar gestations (375 cases, 86.4%) were diploid hydropic abortion. Various trisomy syndromes were found (53 cases, 12.2%), including trisomy 2, 3, 4, 7, 8, 13, 16 and 21. Only 2(0.5%) digynic triploid gestations were detected. Moreover, 4 cases (0.9%) of uniparental disomies (homologous or heterologous) were found. There were 196 cases with histologic diagnostic suspicious of hydatidiform moles were accurate sub-classified. Among them, 59 cases hydatidiform moles were under-diagnosed as diploid hydropic abortions, and 28 cases diploid hydropic abortions were over-diagnosed as hydatidiform moles.Compared with partial moles(PHM), there were no specific histomorphological features between the various types of non-molar gestations and partial moles for definitive diagnostic separation. There was no significant difference in the expression of p57^kip2 among PHM, trisomy and diploid hydropic abortions group (P=0.247). Conclusions STR genotyping can distinguish non-molar gestations from early hydatidiform moles, and efficiently avoid misdiagnosis based only on histological evaluation. Therefore, using STR genotyping, not only can the overdiagnosis of non-molar pregnancy be avoided, but also individualized management can be offered to patients including monitoring of serum hCG.

Objective: To investigate the frequency of KRAS mutation in mucinous epithelial lesions of the endometrium, and analyze the correlation between KRAS mutation and the clinicopathologic features. Methods: The cohort included forty-three cases of mucinous epithelial lesions of the endometrium selected from July 2015 to October 2017 from Beijing Obstetrics and Gynecology Hospital, and 22 control cases. Genomic DNA was extracted from formalin-fixed paraffin-embedded tissue sections. Polymerase chain reaction amplification for KRAS exons 2 and 3 was performed, followed by sequencing using capillary electrophoresis. The Fisher exact test was used to compare the prevalence of KRAS mutation among the different groups. Results: The patients′age ranged from 33 to 77 years [mean (55.12±9.34) years, median 55 years]. None of the eight cases of endometrial hyperplasia with mucinous differentiation without atypia showed KRAS mutation. The frequency of KRAS mutations was 1/10 in endometrial atypical hyperplasia, 1/12 in endometrioid carcinoma, 4/11 in endometrial atypical hyperplasia with mucinous differentiation (EAHMD), 6/15 in endometrioid carcinoma with mucinous differentiation (ECMD) and 8/9 in mucinous carcinoma (MC), respectively. The differences were statistically significant between MC versus EC (P<0.01) and MC versus ECMD (P<0.05). Conclusion The high frequency of KRAS mutation in EAHMD, ECMD and MC indicates that KRAS mutational activation is implicated in the pathogenesis of endometrial mucinous carcinoma.

Objective To explore the association of fibroblast growth factor-23 (FGF23) with abdominal aortic calcification(AAC) and adverse outcomes in maintenance hemodialysis patients.Methods One hundred and fourteen cases of MHD patients were collected prospectively.Serum intact FGF23 was detected by ELISA.Abdomen lateral plain was used as a criteria to determine the abdominal aortic calcification and the abdominal aortic calcification score was counted.Logistic regression analysis was used to determine the risk factors of AAC.Kaplan-Meier analysis was applied to compare the survival rate among different groups and COX regression analysis was used to determine the association of FGF23 and mortality in MHD patients.Results Seventy-six patients present abdominal aortic calcification.The median of AACS was 4.0(0.0,11.0).The median level of FGF23 was 7277.4(2535.0,9990.8) pg/ml.The median follow-up duration was 72.0(67.8,72.8) months.During the follow-up,22 patients (19.3％) died of all-cause death and 17 cases (14.9％) died of cardiovascular diseases.Serum FGF23 level was positively correlated with AAC (r=0.285,P=0.002).Logistic regression analysis showed that longer age (OR=1.059,95％CI:1.020-1.100,P=0.003) and dialysis vintage (OR=I.009,95％CI 1.000-1.017,P=0.039),smoking history (OR=3.010,95％CI 1.177-7.696,P=0.021) and higher FGF23 level(OR=2.831,95％CI 1.010-7.937,P=0.048) were independent risk factors of moderate to severe AAC in MHD patients.Kaplan-Meier survival curves showed that the patients with AACS≥ 5 had significantly higher all-cause mortality(P=0.028) and CVD mortality (P=0.035) than those with AACS ＜ 5.However,the Kaplan-Meier analysis showed no significant difference regarding the level of serum FGF23 with the all-cause and CVD mortality.Cox regression demonstrated that FGF23 was not associated with increased mortality risk,neither in crude nor in multivariate adjusted models.Conclusions Abdominal aortic calcification had a high prevalence in MHD patients.The all-cause and CVD mortality was higher in patients with moderate to severe AAC.FGF23 was an independent risk factor of moderate to severe AAC,but it can't yet be a predictor for the allcause and CVD mortality of MHD patients.

Objective To determine the relationship between serum soluble Klotho (sKL) level and adverse outcome in maintenance hemodialysis (MHD) patients.Methods One hundred and twenty nine cases of MHD patients were collected prospectively.Serum sKL was detected by ELISA.Abdomen lateral plain was used as a criterion to determine the abdominal aortic calcification.The abdominal aortic calcification score (AAC) was calculated.Cox regression analysis was used to determine the risk factor of cardiovascular death (CVD) in MHD patients.Kaplan-Meier showed the relationship between sKL and CVD in MHD patients.Results There were 27 cases (20.9％) of allcause death and 19 cases (14.7％) of cardiovascular death.The median sKL was 612.6(379.2-816.6) nig/L,and log[iPTH] was an independent factor of sKL concentration.Low sKL had high AAC and CVD death rate.Kaplan-Meier method showed that the all-cause death rate was similar between two groups,and CVD death rate increased significantly in low sKL patients (P=0.036).Cox regression indicated that lower sKL level was associated with high CVD death rate [OR=0.352,95％CI(0.127-0.977),P=0.045].After adjustment for the general condition,biochemical indicators,the relationship still existed [OR=0.331,95％ CI (0.117-0.933),P=0.037].In no or mild vascular calcification patients (AAC ≤4),compared with high sKL patients,low sKL patients had no significant difference rate in all-cause mortality.The CVD mortality was significantly higher in high sKL (P=0.035) compared with low sKL.In severe calcification group (AAC ＞ 4),all-cause death and CVD death rates were similar between different sKL groups (P=0.991 and 0.522,respectively).Conclusions Lower sKL has the high CVD death rate and sKL level decreasing is an independent risk factor for CVD death in MHD patients.The lower sKL concentration in MHD patients with no or mild calcification may predict CVD mortality.This study suggests that sKL levels may be helpful in predicting the outcome of patients with MHD.

OBJECTIVE To study the therapeutic effect and the underlying mechanism of asiatico?side on bleomycin-induced rat interstitial pulmonary fibrosis(IPF). METHODS Male Sprague-Dawley (SD)rats were divided into normal control group,bleomycin 5 mg·kg-1 model group and asiaticoside 50 mg · kg-1 group. The model and asiaticoside group were administrated with bleomycin 5 mg · kg-1 to induce IPF,while the asiaticoside group was administrated with asiaticoside 50 mg·kg-1 by gastric perfusion. Hematein eosin(HE)and Masson staining were carried out to analyze the histopathological changes in the lung. Lung homogenates were used to examine hydroxyproline(HYP) content,and serum samples were used to measure the concentration of interferon-γ(IFN-γ),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α). In addition,immunohistochemical methods were used to locate lung transforming growth factor-β1(TGF-β1)and adenosine 2A receptor(A2AR)expression,and Western blotting was used to examine the expression levels of TGF-β1 and A2AR. RESULTS On the 7th,14th and 28th days,the scores of pulmonary inflammation were higher in model group than in control group (P<0.01),and the asiaticoside group showed mitigated alveolitis(P<0.01,P<0.05) compared with model group. Compared with control group,the scores of pulmonary fibrosis in model group were elevated(P<0.01),and the asiaticoside group showed reduced pulmonary fibrosis(P<0.05). On the 14th and 28th days,HYP content in the model group〔1.85±0.10,(2.48±0.18)mg·g-1〕was higher than in the control group〔0.79 ± 0.07,(0.84 ± 0.08)mg · g-1〕(P<0.01),but HYP content in the asiaticoside group〔1.32±0.131,(1.71±0.13)mg·g-1〕was lower than in the model group(P<0.05). IL-4 and TNF-αin the asiaticoside group were lower than in model group(P<0.05),but were higher in the model group than in the control group(P<0.01,P<0.05). The expression level of TGF-β1 protein in the asiaticoside group was lower than in the model group(P<0.05),but was higher in the model group than in the control group(P<0.05). The expression level of A2AR protein in the asiaticoside group was higher than in the model group(P<0.05),but was lower in the model group than in the control group(P<0.05). CONCLUSION Asiaticoside can mitigate bleomycin-induced IPF by inhibiting the expression of IL-4, TNF-αand TGF-β1,and raising the level of A2AR.

OBJECTIVE:To establish the method for the determination of tigecycline (TGC) in human plasma. METHODS:After precipitated by acetonitrile,the plasma sample was determined by LC-MS/MS. Using d9-TGC as internal standard,Kromasil C18 column was used with mobile phase consisted of water (containing 0.05% TFA)-acetonitrile (gradient elution) at flow rate of 0.6 ml/min,column temperature of 40 ℃. The ion transitions were performed under ESI positive MRM model at m/z 586.3→513.2 and m/z 595.3→514.3 for TGC and internal standard,respectively. RESULTS:The linear range of TGC was 25-2 000 ng/ml (r=0.999 8),and lowest quantification limit was 25 ng/ml;intra-day and inter-day RSD was 3.15%-7.23%,and relative error was-4.53%-10.48%. Plasma sample kept stable after 3 times of freezing and thawing cycle,at room temperature for 24 h,in automat-ic sample injector for 24 h and freezing for 42 d (RSD<15%). Plasma concentration of TGC was 0-438.0 ng/ml in one patient with pan-drug resistant bacteria infection(0-12 h after administration). CONCLUSIONS:The developed method is accurate,sensi-tive and specific,and can be used for plasma concentration determination of TGC and pharmacokinetic study.

BACKGROUND:Currently, the enzymatic digestion combined with magnetic activated cel sorting for isolating microvascular endothelial cel s are cumbersome and do harm to cel s. Therefore, how to simplify the isolation and culture of human dermal microvascular endothelial cel s to obtain highly purified endothelial cel s in vitro becomes a hotspot.
OBJECTIVE:To explore a simple and effective cultivation method of microvascular endothelial cel s from diabetic patient skins in vitro, and to detect the cel growth.
METHODS:Diabetic patients with chronic foot wounds after amputation were enrol ed to col ect the limb proximal skin and topical skin around the wound superficial dermal tissue. Human dermal microvascular endothelial cel s were obtained using adherent method and trypsin method, fol oewd by purified utilizing trypsin digestion and repeated attachment method when passage culture.
RESULTS AND CONCLUSION:Human dermal microvascular endothelial cells were obtained successfully, Primary cultured endothelial cells completely adhered to the wall at 24 hours, entered the logarithmic phase at the 10th day, and the cell concentration reached 80%at the 12th-13th day. While the passage cells grew more actively than primary cells, and fully covered the bottom in a“cobblestone”arrangement after 5-7 days of culture. Immunohistochemical staining showed that cultured cells were positive for FVIII and CD31-associated antigens with 100%positive rate. MTT assay showed that cell growth curves of 2, 4, and 5 generations of dermal microvascular endothelial presented the invertedSshape. These results suggest that abundant highly purified human dermal microvascular endothelial cells can be obtained through the adherent method and a small amount of short-term trypsin method.