Henoch-Sch?nlein purpura(HSP)is an IgA-associated leukocytoclastic small-vessel vasculitis, commonly occurred in children.Recent studies have shown that its pathogenesis is closely related to the disturbance of humoral immune response mediated by B cells.CD4 + T cells have the function of help B cells.Each subgroup of CD4 + T cells has unique functions of promoting or regulating immune inflammatory response.There are abnormal changes in the proportion of CD4 + T cells subsets in HSP patients, including the imbalance of Th1/Th2, increasing numbers of Th17 and Tfh cells, together with decreasing Treg cell numbers.The changes are closely related to the occurrence, development and the clinical phenotype of HSP.It is suggested that the detection of CD4 + T cells plays critical roles in guiding the evaluation of HSP, and is expected to provide a new idea for the clinical diagnosis and individualized treatment of HSP.

Objective To construct 2 recombinant plasmids carrying cassette chromosome recombinase C(ccrC)and ccrAB respectively and introduce the plasmids into methicillin-resistant Staphlococcus aureus(MRSA)strain 81/0432,and to observe the precise excision of Staphylococcal cassette chromosome mec(SCCmec)island from bacterial chromosome.Methods ccrC and ccrAB genes were amplified with chromosomal DNAs isolated from MRSA strains 81/0342 and N31S as PCR templates.We constructed recombinant plasmids pSR5C and pSR2AB by cloning ccrC and ccrAB genes into temperature-sensitive plasmid pYI3,after introducing them into MRSA strains 81/0432 and N315 by electroporation.PCR was performed to identify SCCmec excision from the bacterial chromosome.The transformants were serial passaged for 10 days,and then the drug resistance of these rransformants was detected by replica experiment.Results The fragment length of ccrC gene was 1.9 kb,smaller than the fragment length of ccrAB from N315.The recombinant plasmids of pSR5C and pSR2AB were successfully constructed.After these 2 recombinant plasmids were introduced into MRSA strain 81/0342,type-V SCCmec island was excised from the chromosome and formed a closed circular structure in the bacteria.However,type-Ⅱ SCCmec island could be excised only in N31S strain after the expression of pSR2AB.Replica experiment verified that transformed strains of 0342(pSR2AB),0342(pSR5C),and N315 (pSR2AB)were mostly susceptible to ceftizoxim or tobramycin after the excision of SCCmec island.Conclusion cciC could serve as a recombinase as ccrAB,which could mediate precise excision of SCCmec island from the chromosome of type-V MRSA strain.This study shows that type-V SCCmec island is widely disseminated between Staphylococcus aweus strains in community setting.

Objective The study aimed to observe the influence of Puritybile capsules combined with different doses of Compound polyethylene glycol (PEG) in bowel preparation.Methods 120 patients were set into three groups (group A,B and C) randomly (40 patients in each).Group A take the routine Compound polyethylene glycol treatment orally.On the basis of the same routine Compound polyethylene glycol treat-ment,group B added regular dose of Puritybile capsules.Group C took half dose of Compound polyethylene glycol and regular dose of Puritybile capsules.The cleaning effect and adverse reaction were compared among three groups.Results According to Ottawa Scale,the cleaning effects as well as the existing of air bubbles showed no significant difference among three groups.Among which group C was 30％lower than that of group A,10％ lower than that of group B.There was no statistical differences in adverse events.Conclusions Half dose of Compound polyethylene glycol and regular dose of Puritybile capsules can guarantee the ef-fect of bowel cleaning and lower medical expense of patients.

OBJECTIVE:To establish a method for the content determination of L-epicatechin in Actinidiae arguta. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.2%Acetic acid solution(15:85,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 210 nm,column temperature was 25 ℃,and the volume injection was 10 μl. RESULTS:The linear range of L-epicatechin was 10.47-167.52 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;average recovery was 98.07%-101.71%(RSD=1.39%,n=6). CONCLUSIONS:The method is simple, accurate and reliable,and suitable for the content determination of L-epicatechin in A. arguta.

Objective To investigate the dose-response relationship of the treatment temperatures and heating time on human cervical carcinoma hela cells,aiming at providing experimental evidences for clinical hy-perthermia. Methods Hela cells were heated at 37 ～ 70 ℃ in temperature-controlled water baths, the tempera-ture was divided into nine groups,each time was divided into eight subgroups (1 ～ 30 min). The morphology changes of cells after hyperthermia were detected by inverted microscope. Proliferation rates were measured by MTT colorimetric assay. The apoptesis rates were determined by flow eytometric analyse. The levels of prolifera-ring cell nuclear antigen (PCNA) were measured with immunohistochemistry. Results lnereaseing the heating time at the same temperature, or increaseing heating temperature at the same time, the cell proliferation, survival rates and PCNA expression decreased. There was no significant morphological change about cells ,but have small amount of apoptosis and a direct role of the suppression and destruction at 41 ℃ and 43 ℃ group. A large num-ber of cells shrinked to round and a major role for apoptosis at 46℃ group. Cell necrosis was major role at 50 ℃and 55 ℃ group. More than 55 ℃ for necrotic cells. Conclusion With the increase of heating temperature and heating time, its treatment of Hela cells gradually enhance. So combining dose-effect relationship of hyperthermia temperature and time can reach the best therapeutic effects.

ObjectiveTo develop school refusal reason inventory (SRRI)for children and adolescents in China and assess its reliability and validity.MethodsThe primary SSRI was made based on clinical interviews and literatures.Pretest was carried out in a small sample from a clinic.Then the final SSRI was developed after qualitative analysis and item analysis.SRRI,the Screen for Child Anxiety Related Emotional Disorders(SCARED) and Child Depression Inventory(CDI) were administered to school refusers from 7 schools in Shenyang.All the schools were selected from Shenyang City and its countryside by cluster sampling.Some of the students were retested after one month.Descriptive statistics and exploratory factor analysis were carried out to examine the reliability and validity of SRRI based on all the data.Results Item analysis indicated correlation coefficients between all the items and the total marks were higher than 0.3,and they were significant.All the critical ratios of the items were higher than 0.3.The 43 items were divided into six factors ( educational modality,factor of teachers,relationship with classmates,separated anxiety,study attitude and study environment) by exploratory factor analysis.The factor loading values were 0.372 ～0.848.The cronbach's α of each factor was 0.827,0.831,0.759,0.623,0.821 and 0.808.Retest reliability was 0.644 (P ＜ 0.01 ).Its correlation coefficient with SCARED was 0.452 and 0.548 with CDI.ConclusionAccording to Chinese cultural back ground,the SSRI corresponds with psychometric indexes.There are good reliability and validity.It is helpful to understand the reasons of school refusal behavior in children and adolescents.

Objective To investigate the correlation between arachidonate 15-lipoxygenase (ALOX15)gene polymorphism and its genetic predisposition to coronary heart disease (CHD)in Han population of Shaanxi Province so as to provide the basis for early diagnosis and prophylaxis of CHD.Methods The single nucleotide polymorphisms (SNPs)of ALOX15’s rs916055,rs2619112,and rs2664593 were measured by using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF)method in 105 CHD patients (CHD group)and 75 non-CHD patients (control group)who were matched in age and sex.Results The frequencies of genotypes and alleles of SNPs rs916055A/G in CHD group were significantly different from those in control group (P=0.000 1,P=0.000 1).The frequencies of genotypes and alleles of SNP rs2619112A/G in CHD group did not significantly differ from those in control group (P=0.134 2,P=0.143 8).The frequencies of genotypes of SNP rs2664593C/G in CHD group significantly differed from those in control group (P=0.002 7),but the frequencies of alleles were not significantly different (P=0.537 1).Logistic regression analysis indicated that the A allele of SNP rs916055 was an independent risk factor for CHD.Conclusion SNP rs916055 may be related to CHD and its A allele may be the genetic susceptibility gene for CHD.

Objective: To observe the effects of moxibustion at different times on prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) and arginine vasopressin (AVP), in the uterine tissues of rats with dysmenorrhea due to cold-dampness retention, and to explore the differences and possible mechanisms of moxibustion at different times in easing pain in dysmenorrhea due to cold-dampness retention. Methods: Forty-three female Wistar rats were randomly divided into a blank control group (n=7), a model group (n=9), a pre-moxibustion group (n=9), an immediate-moxibustion group (n=9) and a pre-moxibustion plus immediate-moxibustion group (n=9). Rat models of primary dysmenorrhea due to cold-dampness retention were established using (0±1) ℃ ice water immersion method combined with injection of estradiol benzoate for 10 d, followed by injection of oxytocin on the 11th day. Rats in the 3 intervention groups received moxibustion to Shenque (CV 8) and Guanyuan (CV 4), 10 min for each acupoint, once a day. Rats in pre-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling, for 3 continuous days; rats in immediate-moxibustion group were given one time mild moxibustion, immediately after injection of oxytocin on the 11th day during modeling; rats in pre-moxibustion plus immediate-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling till immediately after injection of oxytocin on the 11th day during modeling, for 4 continuous days. The level of PGF2α in the rat uterine tissues was measured by enzyme-linked immunosorbent assay (ELISA), and the levels of PGE2 and AVP in rat uterine tissues were measured by radioimmunoassay. Results: Compared with the blank control group, the levels of PGF2α and AVP, the PGF2α/PGE2 ratio in the model group were significantly increased (P<0.01), and the PGE2 level was significantly decreased (P<0.01) in the rat uterine tissues in the model group. Compared with the model group, the writhing latency was significantly prolonged, the writhing number and the total writhing score were all decreased in the pre-moxibustion group, the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01); the levels of PGF2α and AVP, and the PGF2α/PGE2 ratio were all significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in the rat uterine tissues of the 3 treatment groups. Compared with the pre-moxibustion group, the writhing number and the total writhing score were all decreased in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01), the writhing latency was significantly prolonged in the pre-moxibustion plus immediate-moxibustion group(P<0.01); the levels of PGF2α and PGF2α/PGE2 ratio were significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group. Compared with the immediate-moxibustion group, the writhing latency was significantly prolonged and the writhing number was decreased (all P<0.05), and the total writhing score was decreased (P<0.01) in the pre-moxibustion plus immediate-moxibustion group; the PGF2α level and the PGF2α/PGE2 ratio were significantly decreased (P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the pre-moxibustion plus immediate-moxibustion group. Conclusion: Moxibustion at different times all can produce obvious analgesic effects on dysmenorrhea due to cold-dampness retention in rats, and pre-moxibustion plus immediate-moxibustion ranks the top. The mechanism of this analgesic effect may be via the regulation of abnormal PGF2α, PGE2 and AVP levels, to effectively inhibit the spastic contraction of uterine smooth muscle in dysmenorrhea rat, thereby improving the ischemia and hypoxia in uterus.

Background::One of the significant challenges for cell therapies, such as chimeric antigen receptor (CAR)-T cell therapy, is the poor infiltration of immune cells into tumor tissues. CAR-monocytes/macrophages (CAR-M) are promising therapies because of their enrichment in the tumor microenvironment. Thus, we constructed a novel CAR-M to facilitate the infiltration of T cells and other immune cells.Methods::The suicide gene inducible caspase-9 ( iCasp9) and anti-erb-b2 receptor tyrosine kinase 2 (HER2) CAR elements were transfected into THP1 (an immortalized human monocyte cell line) by lentivirus. The suicide efficiency and specific anti-tumor efficacy were assessed using flow cytometry, inCucyte, and tumor-bearing BALB/c-nude mouse models. The activation of related signaling pathways in CAR-THP1 activation was explored by transcriptome sequencing. Finally, the synergistic therapeutic efficacy of CAR-THP1 combined with RAK cell treatment was demonstrated in tumor-bearing NOD.CB17-Prkdc scid Il2rg tm1/Bcgen mouse models. Results::We developed a novel CAR-THP1, which incorporated iCasp9, CD3ζ, and CD147 intracellular segments, based on the first-generation HER2-CAR backbone. By constructing and comparing a series of CARs with different permutations, CAR-CD3ζ-CD147-iCasp9-THP1 was selected as the optimal combination. CAR-CD3ζ-CD147-iCasp9-THP1 initiated suicide quickly and efficiently under the control of iCasp9 gene, which enabled us to achieve controlled proliferation of CAR-THP1. CAR-THP1 also exhibited robust specific anti-tumor efficacy independently of T cells in vitro and in vivo. Through transcriptional sequencing, we found that CAR-THP1 tended to differentiate into the M1 phenotype and bridged innate and adaptive immunity. A combination of CAR-THP1 and Retronectin actived killer cells (RAKs) showed better therapeutic efficiency, as the metalloproteinases (MMPs) secreted by CAR-THP1 facilitated the degradation of the dense tumor matrix. This further assisted intratumoral infiltration of T cells and augmented the anti-tumor immune response. Conclusion::CAR-THP1 might be effective against HER2-positive tumor cells and has great potential for combination therapy with other immune cells.

Objective:To investigate the effect of calcification on the diagnostic accuracy of the quantitative flow fraction (CT-QFR) derived from coronary CT angiography (CCTA).Methods:A total of 244 patients (471 coronary arteries) who underwent both CCTA and invasive coronary angiography (ICA) for suspected coronary artery disease between 2019 and 2021 were included in the study. All analyses were conducted at the vessel level using CCTA and ICA images, and the morphological and hemodynamic parameters of all enrolled vessels were assessed. The group was divided into severe calcification (206 cases) and non-severe calcification (265 cases) based on whether the arc of lesion calcification was greater than 180°. Subsequently, the two groups were evaluated to the degree of coronary stenosis, the length of the target lesion, the length of calcification, the ratio of the length of calcification, the remodeling index of calcification, the quantitative flow fraction (QFR), the CT-QFR, and the distribution of the involved vessels. Pearson correlation analysis and the Bland-Altman scatterplot were used to analyze the correlation and consistency between CT-QFR and QFR values from different subgroups. The benchmark for coronary ischemia was QFR≤0.80, and the criteria for diagnosing coronary ischemia were CT-QFR≤0.80 and luminal stenosis≥50%, respectively, and the effectiveness of CT-QFR for coronary ischemia was evaluated by plotting the ROC curves in various calcification subgroups.Results:The degree of luminal stenosis, lesion length, calcification length ratio, and calcification remodeling index were substantially higher in the severely calcified group than in the non-severely calcified group (all P<0.05). The results of the Pearson correlation analysis demonstrated a significant association between CT-QFR and QFR in both the severe and non-severe calcification groups ( r=0.85, 95%CI 0.81-0.88, P<0.001; r=0.91, 95%CI 0.89-0.93, P<0.001); in contrast, the Bland-Altman analysis indicated that the CT-QFR and QFR measurements in the severely calcified group exhibited a high level of agreement, with a mean difference of -0.01 (95% limits of agreement -0.22 to 0.20) for measurements in the severely calcified group and 0 (95% limits of agreement -0.15 to 0.16). The specificity, positive predictive value, negative predictive value, and area under the curve (AUC) for the diagnosis of ischaemic lesions by CT-QFR and CCTA alone were lower in the severely calcified group than in the non-severely calcified group, but the difference in AUC between the two groups for CT-QFR was not statistically significant ( P>0.05), and the difference in AUC for the morphological assessment of CCTA was statistically significant. The diagnostic effectiveness of CCTA alone was considerably worse than the specificity and AUC of CT-QFR for the various calcified subgroups for the diagnosis of ischemic lesions (all P<0.001). Conclusions:Severe calcification somewhat affected the diagnosis of ischaemic lesions by CT-QFR, but there was still a high correlation and concordance between CT-QFR and QFR within the severely calcified group, and the diagnostic efficacy was significantly better than that assessed by CCTA morphology alone.