Objective To investgate the expression of Tfh and Th9 in the peripheral blood of rheumatoid arthritis ( RA),and to analyze the relationship between those cells and disease activity,organs of involvement,ect.,and to underly the possible immunological mechananism of RA.Methods Peripheral blood Tfh cells and Th9 cells,define as the CD4+ CXCR5 + ICOS+ and CD3 + CD8+ IL-9+ population,were examined by flow cytometry in 36 patients with RA and 22 case of age-and gender-matched health controls.According to 28-joint Disease Activity Score (DAS28),patients were divied into two groups:high disease activity(n=22),moderate disease activity(n=14).And then to analyzed the correlation among the Tfh,Th9 and clinical data of RA,such as ESR,CRP,RF,the number of tender joints,the number of swollen joints and bone destruction,etc..While to analyzed the correlation between Th9 and Tfh.Results The expression of Tfh (Z=-6.082,P=0.000) and Th9 (0.989±0.498 vs 0.213±0.084,t=13.063,P =0.000 )in PBMCs of RA was significantly higher than normal controls.Meanwhile,the expression of Tfh ( 3.880 ±1.255 vs 2.678±1.022,t=2.990,P=0.005 ) and Th9( 1.181 ±0.523 vs 0.686±0.254,t =4.043,P=0.000) in high activity of group were higher than those moderate.The percentage of Tfh and Th9 in the two groups were all signifieantly higher than controls(P＜0.01 ),There was a positive correlation with Tfh and DAS28 ( r =0.571,P=0.000),ESR ( r =0.375,P =0.029),CRP( r =0.357,P =0.032 ),the number of tender joints ( r=0.598,P =0.000 ),RF ( r =0.421,P =0.023 ) and anti-CCP ( r =0.421,P =0.023 ).There were no relationship with Tfh and course,morning stiffness,the number of swollen joints,bone erosion and the abnormal of EKG.There was a positive correlation with Th9 and DAS28( r=0.461,P=0.005 ),ESR(r=0.347,P=0.042),CRP(r=0.384,P=0.210),the number of tender joints(r=0.341,P=0.042),the number of swollen joints (r =0.347,P=0.038 ) and RF( r =0.379,P =0.025 ) respectively.There were no relationship with Th9 and course,morning stiffness,CCP,the abnormal of EKG,bone erosion.There was a positive correlation with Tfh and Th9 (r=0.727,P =0.000),Conclusion The expression of Tfh and Th9 were increase significantly,which those cells were close relationship with the disease activity and some data,These results indicate that the abnormality of Tfh and Th9 may play an important role in the pathogenesis of RA.

Objective To detect the expression of T helper cells 9 (Th9) in the peripheral blood and the level of interleukin (IL)-9 in the plasma of rheumatoid arthritis (RA) patients,and to analyze the relationship between Th9 and clinical characteristics,and their possible role in the immunological pathogenesis of RA were explored.Methods Thirty-six patients with RA and 22 healthy controls were included.RA patients were divided into high disease activity group (n=22) and moderate disease activity group (n=14).Th9 cells were obtained from RA patients and healthy controls and detected by flow cytometery.The levels of IL-9 in RA patients and healthy controls were detected by ELISA tests.T-test and Spearman's correlation were used for statistical analysis.Results The expression of Th9 and the level of IL-9 of RA patients [(0.99±0.50)％,( 1.06±0.42 ) pg/ml,respectively] were significantly higher than normal controls[ (0.21±0.08)％,(0.69±0.13)pg/ml,respectively,all P＜0.01 ].Meanwhile,the expression of Th9 and the level of IL-9 in high disease activity group [ ( 1.18±0.52 )％,( 1.37±0.47 ) pg/ml ] were higher than those of the moderate disease activity group [ (0.69±0.25)％,(0.94±0.30) pg/ml ].There were positive correlations between Th9 and disease activity score(DAS) 28,erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),the number of tender joints,the number of swollen joints and rheumatoid factor (RF),the level of IL-9 respectively.There were positive correlations between IL-9 and DAS28,ESR,the number of tender joints,and the number of swollen joints.Conclusion High expression of Th9 and level of IL-9 maybe involved in the pat-hogenesis of RA,and is associated with disease activity.These results indicate that the abnormality of Th9 may play an important role in the pathogenesis of RA.

Objective To test the level of cell factor interleukin (IL)-21,CXCL13 in the plasma of patients with rheumatoid arthritis (RA),and to analyze the relationship between Follicular helper T cells(Tfh)and clinic features and discuss the possible immunological pathogenesis of RA.Methods The Tfh cells were obtained from patients and healthy controls (NC) and detected by Flow cytometery.While the levels of IL-21,CXCL13 in patients and NC were measured by ELISA tests.Those analysis were performed by student's t-test,one-way ANOVA,SNK-q test,Chi-square test,Spearman's correlation and multiple linear regression.Results The expression of CD4+CXCR5+ICOS+ cells (Tfh) in PBMCs of RA was significantly higher than normal controls (3.0±1.2 vs 1.1±0.4,P＜0.01).Meanwhile,the three RA groups of patients were divided to low,moderate and high disease activity groups,and the results showed that the expression of Tfh were increased accordingly (1.8±0.7,2.5±0.6,4.0±1.2).The expression of Tfh in the three groups were all significantly higher than that of controls (P＜0.01).There was a positive correlation between Tfh and DAS28,ESR,CRP,TJC,and bone erosion,RF and anti-CCP respectively.The expression of Tfh in those patients who had bone destruction was higher than those with no or mild bone destructions (2.7±1.1vs 3.4±1.3).The expression of Tfh in patients with un-treated RA patients,when compared to those RA patients who were treated appropriately and those who were not treated appropriately,was decreased significantly.The expression of Tfh in appropriately treated RA patients was lower than that without appropriately treatment.The level of IL-21,CXCL13 was decreased in patients with RA in the order of high,moderate,low disease activity and NC.Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,and are closely related to disease activity and bone ersions.The expression of Tfh is decreased after relevant treatment.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of RA.

To screen antitumor active compounds, drug-like or leading compounds from Chinese traditional and herbal drugs.

Objective:To construct a cognitive training program suitable for elderly patients with mild cognitive impairment based on horticultural therapy, so as to effectively slow down the cognitive decline of patients with mild cognitive impairment.Methods:Through searching the Chinese and English database literature of cognitive intervention from July 2000 to July 2020 and field visits to nursing homes, the draft intervention plan was formed. Two rounds of focus group interview were held to consult experts in cognitive impairment and geriatric care, etc., and to revise the intervention plan.Results:In the two rounds of focus group interview, the expert positive coefficient was 100%, the expert judgment basis was 0.84, the expert familiarity degree was 0.84, and the expert authority coefficient was 0.84. In the end, a 10-week cognitive intervention program targeting six cognitive domains -- "visuospatial/executive ability", "memory ability", "language ability", "attention ability", "abstract ability" and "naming ability" was formed, and the implementation steps of the program were improved.Conclusions:The construction process of cognitive training program for patients with mild cognitive impairment based on horticultural therapy theory is rigorous, scientific and feasible, and can be used to guide the cognitive training of patients with mild cognitive impairment.

Objective:To investigate the clinical effect of applying hydrogel dressings in deep partial-thickness burn wounds after dermabrasion and tangential excision.Methods:A prospective randomized controlled study was conducted. From November 2015 to August 2019, 168 patients with deep partial-thickness burns hospitalized in Zibo Sixth People's Hospital met the inclusion criteria. According to the random number table, the patients were divided into hydrogel dressing+vaseline gauze group (84 cases, 67 males and 17 females) and vaseline gauze only group (84 cases, 65 males and 19 females) who were aged (31±16) and (35±17) years, respectively, and were given corresponding treatment after dermabrasion and tangential excision. The dressings were changed every 3-5 days. Autologous skin grafting was performed with split-thickness or medium-thickness skin grafts taken from trunk or thigh if the wounds failed to heal over 21 days or the wounds were less than 21 days but located in the joints, which might affect the functional activities at later stage. After operation, the general condition of the wounds was observed continuously and dynamically till the wounds were healed. The degree of dressing adhesion was evaluated during the first 4 dressing changes after operation, and the degree of pain was evaluated using Numerical Rating Scale (NRS). The wound healing rate on post operation day (POD) 3, 6, and 15 was calculated. The positive proportion of bacterial culture of wound exudates/cleanout fluid on admission and at dressing change on POD 3, 6, and 15 was calculated. The number of dressing changes were recorded, the rate of skin grafting operation was calculated, and the time of complete wound healing was recorded. Vancouver Scar Scale (VSS) was used in 6 months of follow-up to evaluate the hyperplasia of scar. Data were statistically analyzed with analysis of variance for repeated measurement, independent sample t test, Mann-Whitney U test, chi-square test or Fisher's exact probability test, and Bonferroni correction. Results:During the observation period after operation, the wound was moist with less exudates and the wound healed much faster in patients of hydrogel dressing+vaseline gauze group, with the inner dressing being easier to remove with mild pain, while the wounds showed more exudates and slower healing in patients of vaseline gauze only group with the obvious adhesions of inner dressing, stronger pain, and bleeding in the wounds. Compared with those in vaseline gauze only group, the degree of dressing adhesion and pain NRS score of patients in hydrogel dressing+vaseline gauze group decreased significantly during the first 4 dressing changes after operation ( χ 2=52.625, Z=-10.854, P<0.01), the wound healing rate increased significantly at dressing change on POD 3, 6, and 15 ( t=10.347, 41.150, 167.627, P<0.01), the positive proportion of wound exudates/cleanout fluid bacterial culture did not change significantly on admission or at dressing change on POD 3 ( P>0.05) but decreased significantly at dressing change on POD 6 and 15 ( χ 2=15.616, 15.226, P<0.01), the dressing change times was significantly reduced ( t=-11.986, P<0.01), and the rate of skin grafting operation was significantly decreased ( χ 2=35.850, P<0.01). The complete wound healing time of patients in hydrogel dressing+vaseline gauze group was (17.6±2.8) d, significantly shorter than (27.1±3.0) d in vaseline gauze only group ( t=-21.288, P<0.01). During the follow-up of 6 months, the VSS score of scar hyperplasia of patients in hydrogel dressing+vaseline gauze group was significantly lower than that in vaseline gauze only group ( Z=-11.287, P<0.01). Conclusions:Compared with the use of vaseline gauze only, the application of vaseline gauze coated with hydrogel dressing in deep partial-thickness burn wounds after dermabrasion and tangential excision is more effective in moisturizing, significantly reducing the degree of dressing adhesion and patients' pain during dressing change, increasing wound healing rate, decreasing wound infection rate and the rate of skin grafting operation, and reducing the number of dressing changes. It shortens the time for complete wound healing, and effectively relieves scar hyperplasia.

Objective:To explore clustered epidemic of COVID-19 in Liaocheng city and analyze infection status and chain of transmission of the cases.Methods:A joint investigation team of emergency response for COVID-19 epidemic by CDC professional workers of Liaocheng city and district at two levels on January 30, 2020. According to a indicator case from ZH supermarkets, close contacts and related subjects were tracked and screened on February 1, including ZH supermarket employees, family members having contact history with related cases during January 13-26, supermarket clients during January 16-30 and family members of related cases. an epidemiological investigation was carried on and their swab of nose/throat were collected and were sent to Liaocheng CDC laboratory, real-time fluorescence quantitative RT-PCR was used to detect nucleic acids of SARS-CoV-2.Results:a total of 8 437 people were screened during January 30 to February 9, 2020 (120 employees of supermarket, 93 family members, and 8224 clients of supermarket). The epidemic was caused by ZH cases and brought clustered cases in four families. A total 25 cases of SARS-CoV-2 infection, the total infection rate of subjects was 0.30% (25/8 437) with 22 confirmed cases (0.26%, 22/8 437) and 3 asymptomatic patients (0.04%, 3/8 437), asymptomatic patients accounted for 12.00% (3/25) of all infection cases. The infection rates of supermarket employees, family members of confirmed cases and supermarket clients were 9.17% (11/120), 12.90% (12/93) and 0.02% (2/8 224).Conclusions:This was a cluster epidemic caused by one imported case of COVID-19 in a supermarket of Liaocheng city. Prevention and control of cluster epidemic should be focused on chain of community transmission and family cluster cases. It must also be an attention for transmission risk of asymptomatic patients.

Objective To introduce an optimized practical method of making and detecting pipettes for microinjection.Methods Transfer pipette was made from hard glass capillary. We softened the hard glass capillary by rotating it in a spirit-lamp flame,then moved out from the flame and quickly pulled it into two transfer pipettes.After broken by a grinding wheel,the tip of the pipette was fire-polished by quickly touching the flame to make a fine opening.A hard glass capillary (1.0 mm,ouside diametre)was pulled into two holding pipettes by pipette Puller.The pipette shoulder was broken at desired position with a grinding wheel,then the fine pipette tip opening was heated by a microforge and shrinked into a diameter -15 μm.Injection pipette could be made directly from a capillary with filament by Puller.The solution loaded injection pipette and holding pipette were assembled into the micromanipulator and could be checked before use.We transfered both pipettes into the zygotes media drop,touched the holding pipette with the tip of the injection pipette to make a "suitable"opening.Then we switched injection pipette to the mineral oil and applied injection pressure through the injector to check whether the solution could come out of the tip smoothly and at a proper speed.It could be further verified by pronucleus microinjection of zygotes.Results The results showed that the method introduced in this paper could produce suitable pipettes for zygote microinjection.In particular,the method of detecting the opening of the injection pipette was helpful for achieving high efficiency of zygote microinjection.Conclusion The method introduced here to make and detect pipettes for microinjection is very helpful for establishing a standard microinjection manipulation procedure and improving the efficiency of zygote microinjection.

Objective To establish a HPLC method for determining the concentration of voriconazole (VRC) in human plasma.Methods The analysis was conducted using a ZORBAX Eclipse Plus C18 column and the column temperature was 40 ℃.The mobile phase consisted of methanol and water (55∶45).The flow rate was 1 mL · min-1 and the detection was performed at 254 nm.And the internal standard substance was p-Chloroacetanilide.The specificity,lower limit of quantitation (LLOQ),standard curve,precision,accuracy,recovery and stability were investigated.Results Endogenous impurities did not interfere with the determination of the samples in plasma.The standard curve was linear in the range of 0.1-10.0 μg · mL-1 (r =0.999 2),LLOQ was 0.1 μg · mL-1.The RSDs of inter-day and intra-day were all less than 10％ in the plasma and the extraction recovery was 89％-92％.Conclusion This method was simple,sensitive,accurate,efficient and suitable for routine determination of the concentration of VRC in human plasma.

This paper retrieved an updated version of the kidney disease improving global outcomes (KDIGO) guideline and Chinese Medical Association guideline.By combining Micromedex given Thomson grade,we draw a evidence-based evaluation of off-label drug use in pediatric kidney disease.We concluded updated pediatric nephritic disease guidelines.Doctors and pharmacists should cause the attention,and regard off-label uses of drugs normatively and rationally.