OBJECTIVE:To prepare jumazhitong film and establish its quality control method.METHODS:The jumazhitong film former was made with PVOH-124 as the base,the content of the principal agent-dyclonine hydrochloride was determined by HPLC.RESULTS:Good linear relation was achieved when the detection concentration range of dyclonine hydrochloride was 40.8～408?g/ml(r=0.9 998),the average recovery was 101.3%(RSD=1.3%,n=6).CONCLUSION:The preparing technique of jumazhitong film former is simple,its quality is stable and the control method is feasible.

Objective To study the pre-transplant serum level of anti-endothelial cell antibody(AECA)in kidney allograft recipients and its impact on the episode of acute rejection (AR) within 6 months after transplantation. Methods A total of 495 kidney allograft recipients with pre-transplant serum between December 1998 and August 2003 in our center and 40 healthy controls(negative controls)were enrolled in the study.Clinical data including AR within 6 months after transplantation were analyzed retrospectively.The serum AECA level was measured by cyto-ELISA using EA.hy926 cells as substrate,which was shown as the ratio of P (patient)/N (negative control)=(A_(petient)-A_(blank contrnal)/(A_(negative contral)-A_(blank contral).AECA was considered positive when P/N value Was greater than the average A_(negative control)value plus two times the standard deviation.Results Positive rate of AECA was 18.8%(93/495).AECA level in hemodialysis patients who had been on hemodialysis more than 12 months was 1.43±0.37,greater than those less than 12 months(1.27± 0.32,P=0.013)and those of non-dialyzed patients(1.31±0.32,P=0.029).Correlation coeffieient between AECA level and hemodialysis duration was 0.218 (P=0.018).AR incidence in AECA positive recipients was 38.7%,greater than that in AECA negative recipients (23.4%,P=0.002). Incidence of acute T cell-mediated rejection and acute antibody-mediated rejction increased significantly (P=0.035,P=0.002 respectively).Multifactor logistic regression analysis indicated that AECA positive,PRA greater than 1 0%and high CDC level were risk factors of AR with odds ratio of 2.056,1.751 and 1.764 respectively(P=0.004,0.029,0.050). Conclusions The AECA positive in pre-transplant serum indicates the elevated risk of acute allograft rejection.The AECA level increases with prolonged hemodialysis duration.

OBJECTIVE To investigate the protective effect of total saponins from stems and leaves of Panax ginseng (GSLS) on cisplatin (CDDP)-induced kidney damage in mice and its possible mechanism. METHODS Thirty-two male ICR mice were randomly divided into normal control group, CDDP group, and GSLS(150 and 300)+CDDP groups. GSLS was administered to mice by oral gavage once a day for 7 d. On the 7th day, a single injection of CDDP 20 mg·kg-1 was given 1 h after GSLS 150 and 300 mg·kg-1 before GSLS 150 and 300 mg·kg-1 continued to be given for 3 d. Blood urea nitrogen (BUN) and creatinine (CRE) , catalase (CAT) in renal tissue, reduced glutathione (GSH), tumor necrosis factorα(TNF-α) and interleukin 1β(IL-1β) of cisplatin induced mice were detected after 72 h. HE and PAS staining were used to observe the renal histopathological changes;While TUNEL and Hoechst33258 staining were employed to observe apoptosis in kidney tissues. RESULTS Compared with normal control group, CDDP group had a significant reduction in relative body mass (P<0.05), and the level of GSH and CAT in kidney tissues (P<0.05). The level of CRE, BUN, TNF-α, and IL-1βin serum and renal indexes significantly increased (P<0.05, P<0.01), especially BUN and CRE that respectively doubled and quadrupled. CDDP group developed glomerulus swelling, renal tubular expansion and epithelial cell necrosis. Trans?parent tube type of tube cavity appeared, the nucleus pycnosis disappeared, but renal interstitial edema and inflammatory cell infiltration appeared. There was a large amount of glycogen deposition and high expressions of TUNEL positive cells and Hoechst33258 positive cells. Compared with CDDP group, the levels of BUN and CRE in GSLS treatment group significantly decreased (P<0.05, P<0.01) in serum, glycogen deposition was reducted and apoptosis of renal tubular epithelial cells decreased in kidney tissues (P<0.05). The level of TNF-α, IL-1β(P<0.05) and the degree of renal tissue necrosis were significantly reduced (P<0.05) in CDDP+GSLS 300 group, but there was a significant increase in the level of CAT and GSH (P<0.05). CONCLUSION GSLS can protect against mouse kidney injury induced by cisplatin. The mechanism may be related to oxidation, reduced inflammation reaction and resistance to apoptosis.

OBJECTIVETo investigate the effect of genistein on osteoblast proliferation, cellular cycle, apoptosis and differentiation of osteoblasts cultivated under hypoxia conditions.

METHODRat osteoblasts were isolated from calvarias by enzyme digestion and a hypoxic model was established by in a triple-gas incubator. Rat osteoblasts were grouped into the normoxic control group, the hypoxia control group and the hypoxia administration group which was subdivided into Ge-6 group, Ge-5 group and Ge-4 group, to which genistein was administered at doses of 1 x 10(-6), 1 x 10(-5), 1 x 10(-4) mol x L(-1). The cell survival rate, lactic dehydrogenase leakage rate, apoptosis and differentiation of osteoblasts were observed for each group at 3 h after hypoxia, and the gene expression of HIF-1alpha, Bcl-2, Caspase-3 was detected by Real time RT-PCR. Forty-eight hours after hypoxia, osteogenic differentiation markers including alkaline phosphatase activity and nodules were detected.

RESULTCompared with the hypoxia control group, the hypoxia administration group displays a significant increase in the survival rate and a decreased in LDH leakage rate, apoptosis rate and percentage of S + G2 phases. Besides, the mRNA level of HIF-1alpha and Bcl-2 were enhanced, the mRNA level of Caspase-3 was inhibited.

CONCLUSIONGenistein has an effect on protecting osteoblasts from hypoxia.

Alkaline Phosphatase ; metabolism ; Animals ; Apoptosis ; drug effects ; Calcification, Physiologic ; drug effects ; Caspase 3 ; genetics ; Cell Cycle ; drug effects ; Cell Hypoxia ; Cell Survival ; drug effects ; Cells, Cultured ; Genes, bcl-2 ; Genistein ; pharmacology ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; L-Lactate Dehydrogenase ; metabolism ; Osteoblasts ; cytology ; drug effects ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the diagnostic accuracy of CMS50F for screening in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Sixty-four volunteers with suspected OSAHS underwent simultaneous noc?turnal polysomnography (PSG), micromovement sensitive mattress sleep monitoring system(MSMSMS)and CMS50F. The ap?nea-hypopnea index (AHI) detected by PSG and MSMSMS was used as the diagnostic standard for OSAHS. The reliability of CMS50F for monitoring sleep was assessed. Results There was no statistic difference in CMS50F-ODI3 and PSG-AHI be?tween normal, mild and moderate OSAHS groups(P>0.05). The CMS50F-ODI3 was smaller than the PSG-AHI in severe OSAHS patients(P < 0.05). There was a positive correlation between CMS50F-ODI3 and PSG-AHI(r=0.855, P < 0.05). PSG-AHI≥5 events per hour was used as the threshold value to diagnose OSAHS, the sensitivity and specificity of CMS50F were 94.5%and 88.9%. There were no significant differences in CMS50F-ODI3 and MSMSMS-AHI between normal, mild and moderate OSAHS patients(P>0.05). The value of CMS50F-ODI3 was smaller than MSMSMS-AHI in severe OSAHS patients (P < 0.05). There was also a significant correlation between CMS50F- ODI3 and MSMSMS-AHI (r=0.867,P <0.05). MSMSMS-AHI≥5 events per hour was used as the threshold value to diagnose OSAHS, the sensitivity and specificity of CMS50F were 94.5%and 88.9%. Conclusion CMS50F can be used as a portable and reliable device for screening of pa?tients suspected OSAHS.

Objective: To understand the emotional and behavioral problems among preschool children in Ezhou city, and to analyze its association with screen time,to provide a reference for making preventive measures. Methods: 1 720 children were selected from 9 kindergartens in Ezhou by cluster sampling method. Questionnaires were completed by the children’s parents or primary caregivers. Results: High total difficulty score accounted for 9.8%(169/1 720) of all participants,the abnormal proportion of peer communication problems is the highest. The average screen time was (1.95±1.88) hours, among them 2.6% spent more than 2 hours. The average screen time on weekends was (2.84±2.58) hours, among them 19.1% spent more than 2 hours. The results of multiple linear regression analysis showed that weekday, weekend and total screen times was associated with children’s total difficulty score, emotional problems, conduct problems, peer interaction and hyperactivity(P<0.05). Furthermore, with the duration of screen time increased, the susceptibility of emotional and behavioral problems in these four dimensions increased. Conclusion Screen time is associated with emotional and behavioral problems among preschool children. Schools and parents should pay attention to interventions that target family TV viewing practices, and parent-child interaction and communication.

Objective To assess the value of blood oxygen level-dependent magnetic resonance imaging (BOLD-MRI)in diagnosis and prediction of early acute renal transplant rejection.Methods BOLD-MRI was performed in a cohort of 103 patients undergoing cadaver renal transplantation between Dec 2005 and March 2007.Among them,82 recipients had nomlal renal function,21 had biopsy-proved acute rejection.R2* (1/s)measurements were obtained in the medulla and cortex of grafted kidneys. Results R2* values of the medulla were significantly lower in the acute rejection group[R2*=(14.02±2.68)/s]than that in the normally functioning transplants group [R2*=(16.66+2.82)/s],the difference between these two groups was significant (P＜0.01);ROC curve analyses suggested that medullary MR2* values could accurately identify acute rejection in the early post-transplantation period.In the normal functioning transplant group,those with lower medullary R2* values (MR2*＜14.9/s,n=23) had higher acute rejection rates than those with higher medullary R2* values (MR2*＞14.9/s,n=59) in the first 6 months following transplantation,but the difference between these two groups was not significant (17.39% vs 8.47%,P=0.259). Conclusions Mean R2* values in the medullary regions of grafted kidneys with BOLD-MPd may be a non-invasive diadynamic criteria with good sensitivity and specificity,and may be a valuable predictor of early acute renal transplant rejection.

Aim To investigate the possible mecha-nisms of the levels of NO decrease induced apoptosis in human placental trophoblast cells. Methods Human placental trophoblast cells ( HTR-8 ) were cultured in 5 ml DMEM-F12 culture medium with 37℃ 5% CO2 . Then, the old culture medium was discarded and re-placed with 10,100,500,1 000 μmol·L-1 L-NAME, and the group without L-NAME was set as the control group, cultured for 48h. The effects of L-NAME on the survival of cells were detected by methylthiazolyldiphe-nyl tetrazolium bromide ( MTT); the content of NO in cells was tested by nitrate reductive enzymatic;trans-mission electron microscopy, flow cytometry analysis and Annexin-V FITC dyeing were used to test the effects of L-NAME on apoptosis in HTR-8 cells;restore Fe3+ colorimetric assay was applied for detection of to-tal antioxidant capacity ( T-AOC ) , xanthine oxidase for detection of superoxide dismutase ( SOD) activity, and thiobarbituric acid colorimetry for determination of content of MDA. Results Compared with the control group, the survival rate of HTR-8 cells and the levels of NO in 100,500,1 000 μmol·L-1 L-NAME group were significantly reduced(P<0.05,P<0.01). Flow analysis and Annexin-V FITC staining showed that L-NAME could induce cell apoptosis in a dose-dependent manner. The number of cell apoptosis was negatively correlated with the content of NO ( r = -0.5210 ) in HTR-8 cells. Transmission electron microscopy results showed that compared with the control group, the ex-perimental group's cell nucleus shape was irregular, nuclear pyknosis in irregular shape, the chromatin ag-glutination or side the collection, mitochondrial swell-ing or enrichment, crest fracture or dissolved, even vanished, forming the vacuole, especially in 100 μmol ·L-1 L-NAME group, the apoptotic bodies obviously appeared. At the same time, T-AOC, SOD levels in HTR-8 cells decreased ( P <0.05 ) , and the MDA content increased ( P<0.05 ) . The number of cell ap-optosis was negatively correlated with the level of T-AOC ( r= -0.3212 ) , SOD ( r= -0.2779 ) in HTR-8 cells , while positively correlated with the content of MDA(r=0.2807). Conclusion Oxidative stress may play an important role in the levels of NO decrease in-duced apoptosis in human placental trophoblast cells.

Objective:To analyze the incidence of human papillomavirus (HPV) infection in 1 902 patients and to evaluate the efficacy of drug treatment in 266 patients, aiming to provide reference for the treatment of HPV infection.Methods:The subtypes of HPV isolated from 1 902 patients aged 15-86 years visiting the venereology outpatient clinic of Tianjin Medical University General Hospital from October 2019 to May 2021 were identified by polymerase chain reaction-reverse dot blot hybridization. Drug treatment efficacy in 266 patients of them was retrospectively analyzed.Results:The overall incidence of HPV infection in the 1 902 patients was as high as 53.84% (1 024/1 902). It was 52.60% (689/1 310) in males and 56.59% (335/592) in females. There was no significant difference in the incidence between males and females ( P>0.05). The most common HPV genotype in males and females was HPV6 [15.27% (200/1 310) and 21.96% (130/592)], followed by HPV16 [10.61% (139/1 310) and 9.46% (56/592)], HPV11 [9.31% (122/1 310) and 8.61% (51/592)], HPV52 [6.79% (89/1 310) and 8.95% (53/592)] and HPV43 [5.64% (87/1 310) and 8.45% (50/592)]. The majority of HPV-positive patients were aged between 20 and 39 years. There were 476 cases (25.03%, 476/1 902) of single-type infection and 548 cases (28.81%, 548/1 902) of multiple infection. The incidence of multiple infection was higher than that of single-type infection ( P<0.05). The incidence of multiple infection in females was higher than that in males ( P<0.05). Among the 266 patients, 106 were treated with Paiteling, a traditional Chinese medicine (TCM) preparation, and 68 of them tested negative (64.15%) after treatment. Fifty-eight patients were treated with recombinant human interferon α2b and 22 of them (37.93%) tested negative after treatment. Twenty out of the 56 subjects treated with imiquimod tested negative after treatment. Eight out of the 46 patients without treatment also turned negative. Conclusions:The incidence of HPV infection in the 1 902 patients visiting the venereology outpatient clinic was very high, and most of them were young adults. Multiple infection was more common than single-type infection. Topical application of drugs such as Paiteling, recombinant human interferon α2b and imiquimod was effective in treating HPV infection.

Objective:To explore the neuroprotective effect and molecular mechanism of sulforaphane (SFN) on acute carbon monoxide poisoning (ACOP) in rats.Methods:A total of 135 healthy adult male Sprague-Dawley (SD) rats were randomly divided into normal control group, ACOP model group, and SFN intervention group, with 45 rats in each group. The ACOP animal model was reproduced using carbon monoxide (CO) inhalation in a hyperbaric oxygen chamber, while the normal control group was allowed to breathe fresh air freely. The rats in the SFN intervention group received intraperitoneal injection of SFN at a dose of 20 mg/kg once daily starting 2 hours after CO poisoning and continuing until euthanasia. The normal control group and the ACOP model group received equivalent volume of saline injection. Three rats from each group were sacrificed 1 day after intervention to observe the changes in the ultrastructure of neuronal mitochondria in brain tissues under transmission electron microscopy. Six rats from each group were evaluated for cognitive function using neurobehavioral test 7 days after intervention. Brain tissues of 6 rats in each group were collected 1, 3, and 7 days after intervention, and the expressions of phosphorylated-adenosine monophosphate-activated protein kinase (p-AMPK), mitofusin 2 (MFN2), and dynamin-related protein 1 (DRP1) were detected using immunohistochemistry staining and Western blotting. Linear regression analysis was performed to assess the correlations between the expression levels of above proteins.Results:In the normal control group, the rats did not exhibit any abnormalities in cognitive function or the ultrastructure of neuronal mitochondria in brain tissues. ACOP induced cognitive impairment and ultrastructural injury to neuronal mitochondria in rats. However, SFN significantly improved cognitive function in poisoned rats and mitigated the extent of neuronal mitochondrial damage. Over poisoning time, the expression levels of p-AMPK and MFN2 in the brain tissues of ACOP rats were gradually decreased, while the expression level of DRP1 was gradually increased. Compared with the normal control group, the ACOP model group showed significant differences in the expressions of p-AMPK, MFN2, and DRP1. After SFN intervention, the expression levels of above proteins were significantly reversed. Compared with the ACOP model group, the SFN intervention group exhibited a marked increase in the expressions of p-AMPK and MFN2 [p-AMPK positive expression ( A value): 0.226±0.003 vs. 0.177±0.033, p-AMPK protein (p-AMPK/GAPDH): 1.41±0.05 vs. 0.89±0.05, MFN2 positive expression ( A value): 0.241±0.004 vs. 0.165±0.007, MFN2 protein (MFN2/GAPDH): 1.33±0.04 vs. 0.79±0.03, all P < 0.05], along with a significant decrease in DRP1 expression [DRP1 positive expression ( A value): 0.103±0.002 vs. 0.214±0.011, DRP1 protein (DRP1/GAPDH): 1.00±0.03 vs. 1.50±0.03, both P < 0.05]. Linear regression analysis revealed a strong negative linear correlation between DRP1 protein expression and MFN2, p-AMPK protein expressions ( R2 values were 0.977 and 0.971, both P < 0.01), and a positive linear correlation between p-AMPK protein expression and MFN2 protein expression ( R2 = 0.985, P < 0.01). Conclusion:SFN can help maintain neuronal mitochondrial homeostasis by activating the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, thereby alleviating neuronal injury caused by ACOP.