Objective To investigate the effect of cigarette smoke exposure on the interleukin-17-producing CD4+T helper lymphocyte( Th17 ) in peripheral blood and lungs of cigarette smoke-induced emphysema mice. MethodsForty male BALB/c mice were randomly divided into 4 groups, including a control group C12, a control group C24, a smoke-exposure group S12 and a smoke-exposure group S24, 10 mice each group. Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI). The concentration of IL-17, IFN-γ and TNF-α in bronchoalveolar lavage fluid(BALF) and the concentration of IFN-γ and TNF-α in lung homogenates were measured by enzyme-linked immunosorbent assay (ELISA). The proportion of CD4+IL-17+ Th17 cells in peripheral blood and lungs of mice was determined by flow cytometry. The mRNA expressions of RORγt and IL-17 in peripheral blood and lungs were evaluated by real-time PCR. ResultsLm and DI were significantly higher in S12 and S24 groups [(39.19±3.51)μm vs (46.87±7.16) μm, 39.13±1.57 vs 45.16±3.13]than that in C12 and C24 groups [(32.60± 3.21 ) μm vs (32.38±3.73) μm, 28.23± 1.62 vs 28.86±2.07], all P＜0.05. IL-17, IFN-γand TNF-α in BALF of S12 and S24 groups [(119.72±10.72) ng/L vs (296.40±14.00) ng/L,(129.7±22.2) ng/L vs(251. 1±62.4) ng/L, (17.35±1.60) ng/L, (36.35±1.43) ng/L]were higher than those in C12 and C24 groups [(52.06±4.70) ng/L vs (51.89 ±6.82) ng/L, ( 85.8 ±26.8) ng/L vs ( 88.9 ± 11.5 ) ng/L,(6.41 ±0.90) ng/L vs (5.85±0.92) ng/L], IFN-γ and TNF-α in lungs of S12 and S24 groups [( 1124.3±174.4) ng/L vs (1342.7±206.1) ng/L, (77.2±13.7) ng/L vs (101.7±19.0) ng/L, (129.7±22.2)ng/L vs (251.1 ±62.4) ng/L]were higher than those in C12 and C24 groups[(946.2±81.9) ng/L vs (1027.2±188.3) ng/L, (62.1±16.1) ng/L vs(64.4±15. 1) ng/L], all P＜0.05. The percentage of Th17 cells in lungs of S12 and S24 groups [(3.27±1.12)％ vs (7.19±2.24)％, ( 1.96±0.61 )％ vs (3.82±1.26)％]was significantly increased as compared with that of C12 and C24 groups [(1.80± 0.75)％ vs (1.99±0.59)％], all P＜0.05. And the percentage of Th17 cells in peripheral blood of S12 and S24 groups [(1.96±0.61)％ vs ( 3.82± 1.26)％]was also significantly higher than those in C12 and C24 groups [(0.90±0.37)％ vs (0.97±0.32)％], all P＜0.05. In peripheral blood and lungs,the mRNA levels of RORγt and IL-17 in S12 and S24 groups were higher than in C12 and C24 groups, all P＜0.05.Moreover, the frequency of Th17 cells in peripheral blood and lungs had a positive correlation with Lm and DI( r=0.706-0.772 ,all P＜0.05 ). ConclusionAn up-regulation of proportions Th17 in peripheral blood and lungs of cigarette smoke-inducsed emphysema mice were detected. It may be involved in the amplified and persistent inflammation.

Objective To evaluate the expression of Tc17 in a cigarette smoke-induced mice model of emphysema.To explore the probable mechanisms about how Tc17 cells to elevate in lungs of mice.Methods Forty male Balb/c mice were randomly divided into four groups,including control group ( 12 weeks,C12),control group (24 weeks,C24),smoke-exposure group (12 weeks,S12) and smoke-exposure group (24 weeks,S24 ),10 mice each group,Emphysema of mice was observed by HE pigmentation.Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI).The proportion of CD8+ IL-17 + Tc17,CD8+ IL-17 + CC chemokine receptor type 6 ( CCR6 ) + and 6CCR6 + Tc17 cells in lungs of mice was determined by flow cytometry.The mRNA expressions of retinoidrelated orphan nuclear receptor(RORγt) and IL-17 were evaluated by real-time PCR.The levels of IL-1 β,IL-6,IL-23,transforming growth factor β (TGFβ) and CC chemokine ligand 20 (CCL20) were tested by ELISA.Correlations among these indexes were analyzed.Results Lm and DI were significantly higher in S12 and S24 than in C12 and C24,S24 in particular (t value 4.378-15.188,all P ＜ 0.05).The percentages of Tc17 in S12 and S24[(9.28 ± 1.12)%,( 13.13 ±3.56)%]was significantly increased as compared with that in C12 and C24[(2.40 ±0.60 )%,(2.64 ±0.96 )%],S24 in particular.The mRNA levels of RORγt and IL-17 in S12 and S24 were higher than in C12 and C24,S12 and S24 in particular.There was significant difference (all P ＜0.05 ).The frequency of Tc17 cells had a positive correlation with Lm and DI ( r value were 0.734 and 0.884 respectively,P ＜ 0.01 ).The percentages of CD8+ IL-17 + CCR6 +T cells and CCR6 + Tc17 were significantly elevated in S12 and S24 compared to C12 and C24,S24 in particular (all P ＜ 0.05 ).There was positive correlation between Tc17 cell ratio and CCL20 levels( r =0.899,P ＜0.01 ).The levels of IL-1 β,IL-6,IL-23 and TGFβ in S12 and S24 were significantly increased as compared with that in C12 and C24.There was significant difference (all P ＜0.05).Meanwhile,the frequency of Tc17 cells had a positive correlation with IL-1β,IL-6,IL-23,and TGFβ.Conclusions An up-regulation of proportions Tc17 in lungs of cigarette smoke-induced emphysema mice were detected.The CCR6/CCL20 axis and the increased IL-1β,IL-6,IL-23 and TGFβ probably contributed to this up-regulation.

Objective To investigate the osteogenesis capability of different frequency extracorporeal shock wave.Methods 39 rabbits received different frequency extracorporeal shock wave at the middle potion of tibia for 3 or 7 days,these rabbits were then sac rificed and the tibia bones were collected to process for HE and toluidine blue staining,the pathological changes were observed under the light microscope.Results After different frequency extracorporeal shock wave treatment,the typical periosteal reaction were observed,external periosteum bleeded and thickened but there was no reaction at internal periosteum,marrow cavity opened and fibrosed.the osteoblast-1iking cell proliferated,however,no cartilage cells were observed;The rabbits received 7 days shock wave treatment showed more severe reaction than those for 3 days.The shock wave at lower frequency showed more severe reaction than higher frequency.Conclusion shock wave induced osteogenesis through the periosteal reaction of external periosteum;the osteogenesis capability of different frequency extracorporeal shock wave were affected by the frequency.Higher frequency of shock wave was not the ideal way to promote osteogenesis.

Objective To explore the influence of elevating the oxygen pressure on articular chondrocytes in vitro.Method A hydrogen peroxide induced human articular chondrocyte damage model was established.Then the articular chondrocyte viability was detected using the CCK-8 kit.Collagen Ⅱ(COL Ⅱ),The expression levels of aggrecan (ACAN),matrix metalloproteinase 13 (MMP13) and adisintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) were detected using the realtime PCR and Western blotting.Result The viability of articular chondrocytes improved at 12 h but decreased at 24 h after the stimulation of hydrogen peroxide.Twenty-four hours later,the average expression level of COL Ⅱ and ACAN decreased(P＜0.05),while that of MMP13 and ADAMTS5 elevated(P＞0.05).Conclusion Hydrogen peroxide induced elevation of the extracellular oxygen pressure can influence the synthesis and degradation of the articular chondrocyte extracellular matrix.

Objective: To explore the diagnostic values of FibroTouch and FibroScan for liver fibrosis in patients with chronic hepatitis B(CHB). Methods: This study enrolled patients with CHB who was accepted liver biopsy at Beijing Friendship Hospital, Capital Medical University between March 2014 to December 2017. FibroTouch and FibroScan were performed among these patients at same time. Liver stiffness measurement(LSM), optimal cut-off value, receiver operating characteristic(ROC) were compared. Results: In our 103 patients, there were no significantly different between FibroTouch and FibroScan in LSM. The threshold of the optimal cut-off value for FibroTouch and FibroScan were 5.45 versus 5.55 kPa (≥S1), 7.10 versus 6.65 kPa (≥S2), 11.05 versus 9.20 kPa (≥S3), 15.50 versus 15.45 kPa (S4), respectively. The area under the ROC curve for the prediction of the stage1, stage2, stage2, stage 4 of liver fibrosis in these patients were 0.858 versus 0.765 (P=0.54), 0.812 versus 0.801 (P=0.68), 0.863 versus 0.878 (P=0.45), and 1.0 versus 0.99 (P=0.38) respectively. Conclusions FibroTouch and FibmScan have a good consistency in the evaluation of the degree of liver fibrosis in patients with CHB.

OBJECTIVETo perform a comparative assessment of the performance of FibroTouch and FibroScan in patients with hepatitis B.

METHODSA total of 211 patients with hepatitis B, including cases of chronic hepatitis B (CHB) and of compensated cirrhosis, were enrolled for study between June and November of 2013. The patients underwent FibroScan testing (group 1) and FibroTouch testing (group 3), after which the operator examined a time motion ultrasound image from the FibroScan test and located a specific liver portion for focused FibroTouch testing (group 2). The consistency between the two tests' results was investigated by Pearson's correlation analysis, and the difference of liver stiffness between CHB patients and compensated cirrhosis patients was investigated by the two independent samples t-test or Mann-Whitney U test.

RESULTSThe values of liver stiffness were 5.30 (4.30,8.65) in group 1,6.10 (4.70,8.90) in group 2, and 5.70 (4.50, 8.00) in group 3 (all P < 0.05); the Pearson correlation coefficients were all more than 0.8 (P < 0.05) and there was no statistically significant difference found between the results from FibroScan and FibroTouch.The values of liver stiffness were significantly different between the CHB patients and the compensated cirrhosis patients (P < 0.05). The rates of successful detection were 100% for FibroTouch and 97% for FibroScan.

CONCLUSIONFibroTouch and FibroScan have good consistency in the evaluation of the degree of liver fibrosis. FibroTouch has a higher rate of successful detection than FibroScan.

Objective:To identify gene variants and investigate clinical features of nonmuscle myosin heavy chain 9-related disease (MYH9-RD).Methods:In this retrospective study, the data of patients with MYH9-RD admitted to Shenzhen Children′s Hospital from July 2017 to September 2020 were extracted. The gene variants, clinical features and laboratory tests results were summarized.Results:Among the 6 children, 4 were males and 2 were females, aged 4.0 (0.5-7.6) years. Main clinical manifestations included thrombocytopenia (6 cases), epistaxis (3 cases), petechias (2 cases), traumatic hematoma (1 case), and abnormal liver enzymes (1 case). One patient had no family history, and the other 5 cases were pedigrees. Two pedigrees (2 cases) had long-term microscopic hematuria, one pedigree (2 cases) had history of early cataract, and three pedigrees (5 cases) had chronic mild elevation of liver enzymes. Four MYH9 gene variants were found in 12 patients, including c.2104C>T(p.R702C) in exon 17, c.4270G>A(p.D1424N) in exon 31, c.5521G>A (p.E1841K) in exon 39, and c.5797C>T (p.R1933X) in exon 41. According to the family pedigrees analysis, except for the case of variant in exon 17 which was spontaneous mutation with no family history, the other variants were from their father or mother. The complete blood count results showed a decreased platelet number in these patients, and the counting results of the automated hematology analyzer were significantly lower than that of manual counting method ((33.4±17.2) × 10? vs. (60.4±21.0) × 10 9/L, t=-5.83, P<0.05). The examination of the peripheral blood smear revealed the presence of thrombocytopenia with giant platelets and granulocyte inclusion bodies. The MYH9 gene variant (R702C) located at the N-terminus head domain of non-muscle myosin heavy chain ⅡA (NMMHC-ⅡA), which has ATPase activity, led to severe reduction of platelet number (<20×10 9/L) and obscure granulocyte inclusion bodies. However, higher platelet numbers (40×10 9-80×10 9/L) and obvious granulocyte inclusion bodies were observed in patients with tail-position mutations at C-terminus. Conclusions:The clinical phenotypes of MYH9-RD were variable. The mutations in certain regions of MYH9 gene were related to platelet count and granulocyte inclusion bodies. MYH9-RD should be considered in individuals with unknown etiology and persistent thrombocytopenia which is non-responsive to conventional treatment, regardless of family history. Complete blood count and blood smear morphology examinations are the first steps to screen and diagnose the disease. The laboratory should pay attention to the morphological review rules and standardized reports.

OBJECTIVE To investigate the potential mechanism of procyanidin on rats with gingivitis by regulating phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt）/vascular endothelial growth factor （VEGF） signaling pathway. METHODS The rat model of gingivitis was constructed by sewing the neck of the first maxillary molar with silk thread+applying maltose on the gum+feeding with 20% sucrose solution and soft food. Forth-eight model rats were randomly divided into model group， procyanidin group （160 mg/kg）， 740Y-P group （PI3K/Akt signaling pathway activator， 0.02 mg/kg）， and procyanidin+ 740Y-P group （procyanidin 160 mg/kg+740Y-P 0.02 mg/kg）， with 12 rats in each group； another 12 rats were selected as control group； each medication group was treated with corresponding drugs intragastrically or/and intraperitoneally， once a day， for 7 consecutive days. Twenty-four hours after the last administration， the gingival index of rats was measured； the levels of interleukin- 18 （IL-18）， inducible nitric oxide synthase （iNOS） and alkaline phosphatase （ALP） in gingival crevicular fluid， as well as the levels of superoxide dismutase （SOD）， catalase （CAT） and reactive oxygen species （ROS） in gingival tissues of rats were detected； the pathological changes in gingival tissues were observed； the expression levels of PI3K/Akt/VEGF signaling pathway- related proteins in gingival tissues of rats were detected. RESULTS Compared with control group， the gingival tissues of rats in the model group had severe pathological damage，which was manifested as local tissue expansion and congestion， new capillaries， degeneration and loss of collagen fibers and disorder of arrangement， and a large number of inflammatory cell infiltration in the gingival sulcus wall. The gingival index， the levels of IL-18， iNOS， ALP in gingival crevicular fluid， the level of ROS in gingival tissues， the phosphorylations of PI3K and Akt， as well as the protein expression of VEGF in gingival tissues were significantly increased； the levels of SOD and CAT in gingival tissues of rats in model group were significantly decreased （P＜0.05）. Compared with model group， the pathological damage to the gingival tissues of rats in procyanidin group was reduced， and all quantitative indicators were significantly improved （P＜0.05）； 740Y-P could reverse the improvement effect of procyanidin on various indicators （P＜0.05）. CONCLUSIONS Procyanidin may alleviate gingival tissue damage， and improve gingival inflammation and oxidative stress in rats with gingivitis by inhibiting PI3K/Akt/VEGF signaling pathway.

Based on the clinical thinking of combining diseases and patterns， we combined disease identification， pattern differentiation， and constitution identification， and put forward the theory of identifying and treating critical hypertension， which is "examining the symptoms first， identifying the constitutions as reference， and combining the diseases and patterns". Firstly， the starting point of identifying the disease is to examine the symptoms， and those with precise diagnosis and strong specificity will be diagnosed with the disease， while those with relatively broad diagnosis and fuzzy characteristics will be emphasised on identifying constitutions and differentiating patterns. Focusing on the impact of constitution identification on disease identification and pattern differentiation， constitution identification could be the basis when no symptoms to identify， and based on the theory of "constitution-disease correlation" and "constitution-pattern correlation" to improve the understanding of borderline hypertension from the group and individual level， which helps to identify and predict the development of the diseases and patterns； if the symptoms are complicated and difficult to identify， it is necessary to take syndrome as the outline， use the syndrome to unify the disease， and then refer to the constitution to legislate and prescribe medications. This paper summarizes the traditional Chinese medicine clinical differentiation and treatment strategy of borderline hypertension clear and easy to grasp， with a view to provide a feasible and efficient reference for prevention and treatment of borderline hypertension with traditional Chinese medicine.