ObjectiveTo investigate the mechanism of protective effect of Xuebijing injection on vascular endothelial cells in rats with heat stress.Methods Ninety Sprague-Dawley(SD) rats were randomly divided into control, model and Xuebijing injection treatment groups, 30 rats in each group. Heat stress model was reproduced by placing rats in constant temperature box at 40℃, 60% relative humidity for 1 hour, Xuebijing injection group was treated by intraperitoneal injection of Xuebijing 2.5 g/kg, while the control and model groups were treated by intraperitoneal injection of normal saline 2 mL/kg, once a day only in 1 day for both groups. After model establishment, the rectum temperature, heart rate and the mean arterial pressure(MAP) were recorded at 2, 6, 12 hours in each group. At the same time, the rat abdominal aortic blood was collected and serum was separated, the enzyme-linked immunosorbent assay(ELISA) was used to determine the aortic serum levels of lipopolysaccharide(LPS), nuclear factor-κB(NF-κB) and p53, and the prothrombin time(PT), activated partial thromboplastin time(APTT) and D-dimer of venous blood were detected by automatic blood coagulation analyzer(ACLTOP).Results Compared with those in control group, the rectum temperature, heart rate, LPS, NF-κB, p53, PT, APTT, D-dimer were significantly increased, and MAP was obviously decreased in model group(P<0.05 orP<0.01). Compared with model group, the above indexes were improved significantly in Xuebijing injection treatment group at 2 hours〔rectum temperature(℃): 38.02±0.22 vs. 39.32±0.33, heart rate(bpm): 507±14 vs. 562±35, MAP(mmHg, 1 mmHg=0.133 kPa): 98±6 vs. 87±13, LPS(ng/L): 0.65±0.03 vs. 0.82±0.05, NF-κB(ng/L): 1.10±0.04 vs. 1.33±0.05, p53(ng/L): 1.33±0.03 vs. 1.73±0.02, PT(s):15.47±1.03 vs. 20.28±2.01, APTT(s): 40.26±2.46 vs. 47.46±3.51, D-dimer(μg/L): 238.54±8.32 vs. 323.12±8.14,P<0.05 orP<0.01〕.Conclusion Xuebijing injection can correct the disorders of blood PT, APTT, D-dimer via decreasing the secretion of the levels of NF-κB, p53 from vascular endothelial cells in rats with heat stress, thus the integrity of the vascular endothelium can be protected, and LPS entering into the blood stream can be inhibited.

Bacillus subtilis ZC-7 was obtained by implantation with N+ ions beam to B.subtilis AS1.398,and compared with the AS1.398 neutral protease,the enzyme activity of ZC-7 neutral protease was about 1 timeshigher in previous research.A neutral protease was purified from the culture of B.Subtilis ZC-7 by the procedures including amoninium sulfate precipitation,ultrafiltration,DEAE-Sepharose Fast Flow chromatography and Sephadex G-75 chromatography.By multi-step purification,the ZC-7 neutral protease was purified to 78.5 folds and its yield was 27.7%,at last,the specific activity of ZC-7 neutral protease was up to 4.1?105U/mg.Analysed by SDS-PAGE,the purified protease has shown a molecular mass of about 42kDa.The Km for casein hydrolysis was 3.67?10-3?g/ml and the Vmax was 12.21?g/min.The optimum pH and temperature forhydrolysis of casein were 7.0 and 55℃,respectively.This protease was stable up to 40℃ within the pH range of 6.5 and 8.0.EDTA,isopropanol and alcohol nearly inhibited its activity while some ions such as Ca2+,Mg2+,Fe3+ can improve its activity.In addition,it could resist 1 mol/L H2O2.

Objective:To investigate the azole susceptibility of Candida albicans (C. albicans) from vulvovaginal candidosis patients and to analyze the relationship between ERG11 gene mutations in these isolates and azole resistance.
Methods:Three hundred and two clinical isolates of Candida species were collected. Azole susceptibility was tested in vitro in microdilution studies. The ERG11 genes of 17 isolates of C. albicans (2 susceptibles, 5 dose-dependent resistants and 10 resistants) were amplified and sequenced.
Results:Of the 302 isolates collected, 70.2%were C. albicans, of which 8.5%, 3.8%and 4.2%were resistant to fluconazole, itraconazole and voriconazole, respectively. In total, 27 missense mutations were detected in ERG11 genes from resistant/susceptible dose-dependent isolates. Among them, Y132H, A114S, and Y257H substitutions were most prevalent and were known to cause fluconazole resistance. G464S and F72S also has been proved to cause fluconazole resistance. Two novel substitutions (T285A, S457P) in hotspot regions were identified.
Conclusions:Twenty seven mutations in the ERG11 gene were identified in azole-resistant C. albicans isolates, which indicated a possible relation with the increase in resistance to azole drugs and the recurrence of vulvovaginal candidosis. The relationship of two novel substitutions (T285A, S457P) with fluconazole resistance needs to be further verified by site-directed mutagenesis.

Objective To investigate the titer stability of the harvest solution of severe acute respiratory syndrome coronavirus2(SARS-CoV-2)at 2 ～ 8 ℃ and the inactivation effect of β-propiolactone inactivator on the virus.Methods Three batches of SARS-CoV-2 harvest solution(batch numbers:202111001,202111002 and 202111003)were stored at 2 ～ 8 ℃ for 12 d and sampled every 3 d(0,3,6,9 and 12 d)for detection of the titers by Karber method;Three batches of virus harvest solution equilibrated overnight at 2 ～ 8 ℃ were inactivated by adding β-propiolactone at a volume fraction of 1∶4 000 and detected for the titers at different inactivation time points(0,0.5,1,1.5,2,3,4,8,16 and 24 h),of which samples inactivated for 8,16 and 24 h were taken for inactivation verification,and samples inactivated for 24 h were observed by transmission electron microscope.Results The titers of SARS-CoV-2 decreased with the prolongation of storage time at 2 ～8 ℃,which showed no obvious decrease during 0 ～ 3 d,while decreased from the initial 7.75,6 and 7.5 lgCCID_(50)/mL to5.75,4.625 and 6.25 lgCCID_(50)/mL on day 12,indicating that the virus activity showed a gradual decrease trend at 2 ～8 ℃；With the inactivation time,the virus titer decreased continuously and could not be detected after inactivation for 3 h.Transmission electron microscope observation showed that the inactivated virus particles were intact and the spike protein was evenly distributed.Conclusion The virulence of SARS-CoV-2 stored at 2 ～ 8 ℃ was unstable,so the subsequent inactivation and purification process should be carried out as soon as possible;The titer of virus could not be detected after3 h of inactivation,which provided a reference for the determination of the inactivation process.

AIMTo explore the molecular biological mechanism of hemoglobin with high oxygen affinity in Tibetans by determining the sequence of globin cDNA in Tibetans living at high altitude.

METHODSTotal RNA was isolated from human bone marrow samples of three Tibetans who live in Qinghai-Tibet plateau. cDNA fragments coding for alpha and beta genes of human hemoglobin were obtained through RT-PCR and were ligated to plasmid pGEM-T easy vectors, and then the ligation liquid were transformed to Escherichia coli and cloned and sequenced. Nucleotide sequences were compared with GenBank data by BLAST method.

RESULTSsequence of a globin cDNA in Tibetans were the same with the registering globin genes in the GenBank, and Hb Abruzzo (beta143 (H21), His- > Arg) gene mutation, a high oxygen affinity beta globin mutation, was found in one Tibetan' beta goblin coding gene (CAC- > CGC).

CONCLUSIONThis hemoglobin gene mutation may be associated with high altitude adaptation of Tibetans living at high altitude.

Adult ; Altitude ; Asian Continental Ancestry Group ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; RNA ; Sequence Analysis, DNA ; Tibet ; alpha-Globins ; genetics ; beta-Globins ; genetics

OBJECTIVETo investigate the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

OBJECTIVETo evaluate the effect of the modified urethral pull-through procedure in the surgical treatment of posterior urethral stricture or obliteration caused by trauma.

METHODSFrom January 2001 to June 2005, modified urethral pull-through procedures were performed in 36 patients with post-trauma posterior urethral stricture or obliteration, 25 of them with post-operation urethral stricture or obliteration after urethral realignment by traction and the other 11 treated by cystostomy at the time of initial injury. The length of the urethral stricture segment was 1.0-4.5 cm, mean 2.2 cm. The patients were aged from 17 to 59 years (mean 44.5 years).

RESULTSAll the patients were followed up for at least 1 year. Success was achieved in 34 patients, among whom 25 had good voiding and no need of dilation, 6 were cured after 3 - 6 post-operative dilations, 3 required dilation every 2 or 3 months for over a year and 2 failed. No blood transfusion was needed during the operation, and no erectile dysfunction or incontinence occurred after it.

CONCLUSIONThe modified urethral pull-through procedure is effective and simple in the surgical treatment of post-trauma posterior urethral stricture or obliteration, with few complications and no consequent erectile dysfunction or incontinence.

Adolescent ; Adult ; Follow-Up Studies ; Humans ; Middle Aged ; Treatment Outcome ; Urethra ; pathology ; surgery ; Urethral Stricture ; etiology ; surgery ; Urologic Surgical Procedures ; methods ; Wounds and Injuries ; complications

Objective To discuss the molecular mechanism of 18F-fluorodeoxyglucose (FDG) uptake in tumor and to assess its value to identify pathologic type and cancer staging in patients with earlystage nasopharyngeal carcinoma.Methods Forty patients with nasopharyngeal carcinoma of early-stage,including 12 cases with T1 stage and 28 cases with T2 stage, underwent FDG PET imaging.The maximum standardized uptake value ( SUVmax ) and mean standardized uptake value ( SUVmean ) of FDG uptake of each patient were measured and compared between T1 and T2 stage by t-test.The expression of glucose transport protein 1 ( Glut1 ) and hexokinase- Ⅱ ( HK- Ⅱ ) of each case was measured in paraffin sections by streptavidin-perosidase (SP) immunohistochemistry.The positive expression rate of Glut1 and HK- Ⅱ was calculated and compared between T1 and T2 by x2 test.Meanwhile, the correlation between the expression of Glut1 or HK-Ⅱ and the SUVmax was tested by Pearson analysis.Results The SUVmax and SUVmean in 40 patients were 9.45 ± 1.87 and 6.04 ± 1.09, respectively.The SUVmax of patients with T1 stage (8.95 ± 1.91 ) was significantly lower (t =4.46, P＜0.001 ) than that of patients with T2 stage (11.55 ± 1.70), and the SUVmean of patients with T1 stage (5.61 ± 1.08) was significantly lower ( t = 6.76, P ＜ 0.001 ) than that of patients with T2 stage (7.98 ± 1.10) too.Among 40 patients, all patients showed positive expression of Glut1 and HK-Ⅱ , and the positive expression rate of Glut1 and HK-Ⅱ was ( 45.2 ± 10.9 )% and ( 68.3 ±9.5)%, respectively.The positive expression rate of Glut1 was (38.4 ±8.1)% in T1 stage and (49.7 ±12.6)% in T2 stage, which displayed no difference (x2 =40.58, P＞0.05), but the HK-Ⅱ positive expression rate showed significant difference (x2 =58.71, P＜0.05) between T1 stage (60.1 ±11.1)% and T2 stage (77.9 ± 14.7 )%.The correlation analysis indicated that there was low-degree positive correlation (r =0.369, P=0.019) between the SUVmax and Glut1 expression, and there was medium-degree positive correlation (r = 0.549, P = 0.001 ) between the SUVmax and HK-Ⅱ expression.Conclusion Expression of Glut1 and HK-Ⅱ was positively correlated with FDG uptake in patients with early-stage nasopharyngeal carcinoma.

Objective To analyze the characteristics of functioning in children with autism spectrum disorder (ASD) based on the ICF-CY, to discuss the approaches of physical activity and exercise rehabilitation for them. Methods The typical disabilities in children with ASD were analyze based on ICF-CY and literature review. The approaches of physical activity and exercise rehabilitation were discussed. Results There were two kinds of behaviors including repetitive behavior and communication impairments. The main functioning of children with ASD included special mental function and motor function in body function and body structure;interpersonal interactions, hand and arm flexibly use, walking and moving in activity and participation. Individualized physical activity plan for children with ASD should promote the development of motor skills, improve relevant functions and promote all-round developments. Conclusion It is useful to analyze the functioning and disability, to develop individualized plans of physical activity and sport rehabilitation, to improve total functioning and to promote all-round developments.