Objective:To explore the clinical manifestations,pathological features,immunophenotype,as well as diagnosis,treatment and prognosis of patients with CD4-CD56+blastic plasmacytoid dendritic cell neoplasm(BPDCN),in order to further understand the rare disease.Methods:The clinical data,laboratory examinations and treatment regimens of two patients with CD4-CD56+BPDCN in the First Affiliated Hospital of Wannan Medical College were retrospectively analyzed.Results:The two patients were both elderly males with tumor involved in skin,bone marrow,lymph nodes,etc.Immunohistochemical results of skin lesions showed that both CD56 and CD123 were positive,while CD4,CD34,TdT,CD3,CD20,MPO and EBER were negative.Flow cytometry of bone marrow demonstrated that CD56,CD123,and CD304 were all positive,while specific immune markers of myeloid and lymphoid were negative.Two patients were initially very sensitive to acute lymphoblastic leukemia or lymphomatoid chemotherapy regimens,but prone to rapid relapse.The overall survival of both patients was 36 months and 4 months,respectively.Conclusion:CD4-CD56+BPDCN is very rare and easily misdiagnosed as other hematological tumors with poor prognosis.Acute lymphoblastic leukemia or lymphomatoid therapy should be used first to improve the poor prognosis.

AIM To isolate and identify the dominant strains in the fermentation process of the HuaFengDan YaoMu,and to investigate the effects of different temperatures and pH values upon their growth.METHODS During the 0-6 weeks fermentation process of HuaFengDan YaoMu in the selective medium,the cultured bacteria,the molds and the yeasts had the colonies counted,and the dominant strains isolated and purified,particularly the dominant bacteria and fungi determined by 16S rDNA and 26S rDNA sequences.The optimal growth temperature and pH value of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae were investigated respectively.RESULTS HuaFengDan YaoMu found its bacterial counts increased from 0 to 4 weeks and stabilized from 4 to 6 weeks;its yeast counts increased during 0-3 weeks,and decreased continuously during 4-6 weeks;and its mold counts remained unchanged at 0 week,but increased steadily from 1 to 4 weeks,and decreased at 5 to 6 weeks of the fermentation.In the fermentation process of HuaFengDan YaoMu,the dominant strains of Bacillus aryabhattai,Debaryomyces hansenii and Aspergillus oryzae,identified and screened out by the phylogenetic tree,adapted best to the growth temperature of 35℃,25-30℃and 35℃,at pH value of 8,6 and 7,respectively.CONCLUSION In the fermentation process of HuaFengDan YaoMu,the dominant microorganisms of Bacillus aryabhattai,Debaryomyces hansenii,Aspergillus oryzae are identifiable and separable.

ObjectiveTo explore the regulatory effect of Gouqi chewable tablets on innate and adaptive immunity in normal mice and its antioxidant activity in vitro and in vivo. MethodThe effects of low-， medium-， and high-dose groups （0.25， 0.5， 1.5 g·kg^-1） on the immune function of normal mice were observed by carbon clearance test， immune organ index test， serum hemolysin test， ConA-induced splenic lymphocyte proliferation test， and natural killer cell （NK cell） activity test. The effects of Gouqi chewable tablets on the antioxidant capacity in vivo were determined by detecting the content of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and malondialdehyde （MDA） in mice serum. The in vitro antioxidant activity of Gouqi chewable tablets was detected by 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） diammonium salt （ABTS）， 2，2-diphenyl-1-picrylhydrazyl （DPPH）， and hydroxyl radical scavenging tests. ResultCompared with the blank control group， the low-， medium-， and high-dose groups of Gouqi chewable tablets improved the viability of NK cells， the proliferation of splenic lymphocytes， and the level of serum hemolysin antibody in mice （P<0.05）. The high-dose group increased the thymus index， spleen index， and phagocytic function of macrophages （P<0.05， P<0.01）. As compared with the blank control group， the activity of GSH-Px in mice serum in the medium-dose group was increased （P<0.05）， and the content of MDA in mice serum in the high-dose group was decreased （P<0.05）. In in vitro antioxidant tests， the median inhibitory concentration （IC₅₀） values of Gouqi chewable tablets were 1.64±0.20， 2.04±0.03， and 10.27±0.03 g·L^-1 by the DPPH， ABTS， and OH^- free radical method， respectively. Those results indicated that Gouqi chewable tablets have good antioxidant effects in vitro. ConclusionGouqi chewable tablets can enhance the immune function of mice with good antioxidant effects.

Objective:To observe the clinical efficacy of Qingzhenfang for plasmoby （chronic urticaria）， and to investigate its effect on cellular immune function. Method:One hundred and thirty-two cases patients were divided into control group and observation group evenly according to random number table. The 60 patients in control group finished the study because of 6 cases of dropout， loss of follow-up and withdrawal， and 62 patients in observation group finished the study. Patients in both groups got Yiebastine tablets， 10-20 mg/time， 1 time/day. Patients in control group additionally got Piminxiao capsule， 4 grains/time， 3 times/day， while patients in observation additionally got Qingzhenfang， 1 dose/day. The treatment continued for 8 weeks in both groups. Before the treatment， and at the second， fourth， and eighth week after treatment， scores of urticaria activity for 7 days （USA7） and total symptom score （TSS） were graded. Before and after treatment， scores of chronic urticaria quality of life scale （CU-Q2oL） and syndrome of rheumatic fever were graded. A follow-up of 3 months was conducted for the patients whose score of USA7 was less than 7 to record the recurrence. Complement 3 and 4 （C3， C4）， CD4⁺， CD8⁺ cells were detected， and Th17/ CD4⁺ and Treg/ CD4⁺， CD4⁺/CD8⁺ and Th17/Treg were calculated. Levels of peripheral blood interleukin-10 （IL-10）， IL-17 and IL-23 were detected， and safety was evaluated after the treatment. Result:At the second， fourth and eighth week after the treatment， scores of USA7， TSS， CU-Q2oL and syndrome of rheumatic fever in observation group were lower than those in the control group （P<0.01）. Levels of C3， C4， CD4⁺， Treg， CD4⁺/CD8⁺and IL-35 in observation group were higher than those levels detected in control group （P<0.01）， while levels of CD8⁺， Th17， Th17/Treg， IL-10， IL-17 and IL-23 were lower than those in the control group （P<0.01）. Recurrence rate was 25.58% （11/43） in observation group， lower than 48.48% （16/33） in control group （χ2=4.276， P<0.05）， and the clinical efficacy in observation group was superior to that in control group （Z=2.021， P<0.05）. Conclusion:Yaoyi Qingzhenfang can control the degree of disease and improve the quality of life for patients with chronic urticaria， with superior clinical efficacy. In addition， it can reduce recurrence rate， increase the levels of C3， C4， regulate cellular immune function， and reduce immune inflammatory response， so it is worthy of further clinical research and use.

Objective:To establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages， and to control the quality of this formula from the biological activity level. Method:The phagocytic and inflammation models of RAW264.7 macrophages were established， the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6 （IL-6） secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang， and the biological limit was searched. Result:The optimal inoculation density of RAW264.7 macrophages was 3×10⁵ pcs/mL， and the concentration of lipopolysaccharide （LPS） was 1 mg·L^-1 after treatment for 24 h. When the concentration was 500 mg·L^-1， water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages， and at this concentration， the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted， the phagocytic index was >113%. In addition， the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS， the inhibitory rate was >45%. Conclusion:Combined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang， this study establishes an in vitro biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages， and 500 mg·L^-1 was confirmed as the limit concentration. Under the limit concentration， Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS， which can be judged as qualified.

Aim To establish three kinds of DNA damage models of mouse testicular spermatogonia cell line GC-1 cells and analyze their similarities and differences. Methods GC-1 cells were treated with UVB irradiation, D-galactose(D-Gal) or bleomycin (BLM), respectively. Then the expression and localization of 'Y-H2AX were detected by Western blot and immunofluorescence, the expression and localization of 8-OHdG were measured by immunofluorescence, and the expression levels of p-p53 and p21 were measured by Western blot. Results The expression of -y-H2AX in GC-1 cell reached to the peak 4 h after UVB irradiation and 6 h after D-Gal stimulation, whereas -y-H2AX expression gradually increased after BLM stimulation, and the higher the concentration of BLM,the shorter the time to reach the peak. The results of immunofluorescence showed that 8-OHdG expression was observed in the nucleus and cytoplasm of GC-1 cells after UVB irradiation and BLM stimulation, and the longer the culture time, the more the expression in the nucleus. In contrast, the expression of 8-OHdG was observed in the cytoplasm and reached the peak at 6 h in the D-Gal stimulated GC-1 cells. After UVB irradiation, the protein expression levels of p-p53 gradually increased, while p21 protein expression appeared later than that of p-p53; in the D-Gal stimulated GC-1 cells, the protein expression levels of p-p53 reached the peak at 6 h, and p21 protein expression reached the peak at 12 h; after low concentration BLM stimulation, the protein expression levels of p-p53 and p21 continuously increased, and after high concentration BLM stimulation, the protein expression levels of p-p53 and p21 reached its peak at 2 h, then decreased at 4 h. Conclusions Three kinds of DNA damage models of GC-1 cells are successfully established, and the DNA damage in GC-1 cells treated with D-Gal is mild, while the DNA damage in GC-1 cells treated by UVB irradiation and BLM stimulation is more severe.

Aim To compare the effects of BoneCeramic and Bio-Oss on the proliferation, adhesion and osteogenic differentiation of dog bone marrow mesenchymal stem cells (DBMSCs) in vitro. Methods DBM-SCs were isolated and cultured in vitro and identified for osteogenic and adipogenic differentiation. CCK-8 was used to detect the effects of BoneCeramic and Bio-Oss bone meal extracts on the vitality of DBMSCs. SEM was used to observe the adhesion of DBMSCs on the surface of bone meal. ALP was used to quantitatively detect the osteogenic differentiation of DBMSCs on the surface of bone meal, and the relative expressions of OPN, OCN, RUNX-2, and ALP genes were detected by RT-PCR. Results The cells isolated and cultured from dog bone marrow conformed to the characteristics of mesenchymal stem cells. CCK-8 results showed that the stock solutions of Bio-Oss and Bone Ceramic had no obvious cytotoxic effect on DBMSCs. SEM showed that DBMSCs were distributed more widely on the surface of BoneCeramic than that of Bio-Oss. ALP quantitative detection showed that DBMSCs on the BoneCeramic surface were more conducive to its osteogenic differentiation than Bio-Oss, and it also increased the expression of OPN, OCN, RUNX-2 and ALP genes. Conclusions In vitro studies show that both Bio-Oss and BoneCeramic have good biocompatibility. Compared with Bio-Oss bone meal, BoneCeramic is more conducive to the osteogenic differentiation of DBMSCs.

Objective:To investigate the serum level of low density lipoprotein cholesterol (sdLDL-C) in elderly patients (age≥65 years) with type 2 diabetes mellitus (T2DM) and the its predictive value in evaluatingthe risk of cardiovascular and cerebrovascularevents in elderly patients with T2DM.Methods:In this retrospective study,386 elderly patients with T2DM were collected from December 2014 to December 2016, the averageage was (72.7±5.4) years old, including 269 males and 117 females; 92 of whom had cardiovascular events during follow-up, the averageage was (72.9 ± 5.2) years old, including 65 males and 27 females, and serum sdLDL-C level was detected. Meanwhile, biomarkers such as lipoprotein (a), apolipoprotein B, apolipoprotein AI and hypersensitive c-reactive protein were analyzedin all the patients.Univariate and multivariate factors were used to analyze the relationship between each variable and the occurrence of cardiovascular and cerebrovascular events. The correlation between LDL-C, sdLDL-C of all subjects with age and other lipid indexes were analyzed. ROC curve was used to determine the predictive value of sdLDL-C elevation for cardiovascular and cerebrovascular events in elderly patients with T2DM.Results:The levels of LDL-c, sdLDL-C, non-HDL-C, GLU, HbA1c and ApoB were significantly higher in theCardio-cerebral vascular event group ( t=3.26, 3.46, 2.91, 2.47, 4.03, 3.00, P<0.05). While the levels of apolipoprotein AI was significantly lower than those in theNon-cardio-cerebral vascular event group ( t=-2.39, P<0.05). Cox regression analysis showed that sdLDL-C per 10 mg/dl was independently correlated with the risk of cardiovascular and cerebrovascular events( HR 1.281, 95 %CI 1.225-16.032, P<0.01)after adjusted for age. SdLDL-C was positively correlated with TG, non-HDL-C and ApoB ( r=0.323, 0.588, 0.623, P<0.01) and was negativelycorrelated correlation with age, HDL-C and apolipoprotein AI ( r=-0.363, -0.301, P<0.01), ROC curve analysis showed that sdLDL-C had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.736, 95 %CI 0.554 9-0.918 2, P=0.003), while HbA1c also had a strong ability to predict the occurrence of cardiovascular and cerebrovascular events(AUC=0.725, 95 %CI 0.524 3-0.927 3, P=0.006). Combined sdLDL-C with HbA1had the strong ability to predict the occurrence of cardiovascular and cerebrovascular events (AUC=0.837, 95 %CI 0.711 4-0.973 5, P=0.001). Conclusions:The elevation of serum sdLDL-C in elderly patients with T2DM wasa significantly independentrisk factorof cardiovascular and cerebrovascular events. Serum sdLDL-C had a higher clinical value than LDL-C, which was expected to be the most effective predictor of lipid profile in riak assessment of cardiovascular and cerebrovascular events in elderly patients with T2DM.

Objective:Lung ultrasound (LUS) has been used in the diagnosis of neonatal respiratory distress syndrome(RDS) successfully, but there have been no multicenter prospective studies to verify its reliability or determine how to grade RDS with LUS findings.This study aimed to discuss the necessity and feasibility of using LUS findings to determine RDS grades through a multicenter prospective study.Methods:Every researcher participated in the National Neonatal Lung Ultrasound Training Course and receiving 3-6 months of lung ultrasound system training at the National Neonatal Lung Ultrasound Training Center.Patients between June 2018 and May 2020 who met the RDS ultrasound diagnostic criteria and had full available clinical data were included in this study.The LUS examination was completed immediately after the patients were admitted to the hospital.Some of them also underwent chest X-ray examination.Arterial blood gas analysis was completed immediately before or after the LUS ultrasound examination.RDS grading was performed according to the LUS findings and whether the patient had serious complications.Results:A total of 275 qualifying cases were included in this study, which included 220 premature infants and 55 full-term infants, and the primary RDS occurred in 117 cases (42.5%), and secondary RDS occurred in 158 cases (57.5%). LUS manifestations of RDS patients can be divided into three categories: (1)A ground-glass opacity sign: which could be found among 50 infants when they were admitted to the hospital (that was, at their first LUS examination). Twenty-eight of these infants were considered to have wet lungs and were not sent for special management on admission, but LUS showed typical snowflake-like lung consolidation within 0.5 to 4 hours.Twenty-two of them were given mechanical ventilation with exogenous pulmonary surfactant; Eighteen cases were controlled within 6-12 hours, but the lung lesions became more severe in the other 4 infants (due to severe intrauterine infection). (2)Snowflake-like lung consolidations: the first LUS on admission showed typical snowflake-like lung consolidation involving areas ranging from 1-2 intercostal spaces to 12 lung divisions in 204 cases.Thirty-eight infants among them the lung consolidation only had involvement of 1-2 intercostal spaces at the time of admission; Fifteen of them received invasive respiratory support and recovered within 4-12 hours.Twelve patients received noninvasive respiratory support; Seven of them recovered, while five cases developed severe lung illness.The remaining 11 patients who were not given any form of ventilator support developed severe conditions within 1-4 hours.Thirty of them showed snowflake signs involving 12 lung regions at admission.The remaining 136 patients had lung consolidation degree between the two degree above condition.(3)Snowflake-like sign with complications: Twenty-one patients had severe complications such as pneumothorax, pulmonary hemorrhage or/and persistent pulmonary hypertension of the newborn or large area atelectasis, etc, although snowflake lung consolidation did not involve all lung regions.Conclusion:(1) LUS is reliable and accurate for diagnosing RDS.RDS has the same characteristics on ultrasound for both preterm and full-term infants, both primary and secondary RDS.(2) To facilitate the management of RDS, it is necessary to classify RDS according to the ultrasound findings and the presence of severe complications.(3) Based on the results of this study, it is recommended that RDS can be divided into mild, moderate and severe degrees.The exact standards for grading are as follows: Mild RDS: the early stage of RDS, in which lung consolidation shows as a ground-glass opacity sign on ultrasound; Moderate RDS: lung consolidation shows a snowflake sign on ultrasound, not all of the lung fields are involved; Severe RDS meets one or more of the following criteria: lung consolidation shows as a snowflake sign on ultrasound and all lung regions are involved, or regardless of its degree and extent, lung consolidation has caused serious complications, such as pulmonary hemorrhage, pneumothorax, persistent pulmonary hypertension of the newborn, or/and a large area of pulmonary atelectasis.

To study the protective effects of Wuzi Yanzong recipe on DNA oxidative damage of testis germ cells in natural ageing rats based on Nrf2/HO-1 signaling pathway and base excision repair (BER). In the study, 16-month-old SPF grade male SD rats were randomly divided into three groups, namely ageing model group, and low and high-dose Wuzi Yanzong recipe groups (WZ, 1, 4 g·kg⁻¹). In addition, 2-month-old SD rats were used as adult control group (10 rats in each group). The ageing model group and the adult control group were fed with normal diet for 4 months. WZ groups were given medicated feed for 4 months. After fasting for 12 hours, the rats were put to death. Then, the testes were immediately removed. The vitality of superoxide dismutase (SOD) and malondialdehyde (MDA) content in testis were detected by xanthine oxidase method and thiobarbituric acid (TBA) method. The levels of Nrf2 and 8-OHdG were detected by immunofluorescence. The protein expression levels of Nrf2, HO-1, NQO1, APE1, OGG1 and XRCC1 were detected by Western blot. Compared with the ageing model group, WZ significantly increased the SOD vitality and decreased MDA content of testis. In addition, immunofluorescence results showed that WZ significantly attenuated testicular DNA oxidative damage and improved antioxidant capacity. Such changes were accompanied by the down-regulation of DNA oxidative damage response protein 8-OHdG levels and the up-regulation of Nrf2 levels. Moreover, Western blot results showed that WZ significantly increased the protein expression levels of Nrf2, HO-1 and NQO1 of the testis germ cells, when compared with ageing model group. In parallel, the protein expression levels of APE1, OGG1 and XRCC1 were significantly decreased. In conclusion, WZ improves ageing-related DNA oxidative damage via Nrf2/HO-1 and BER pathways.