Objective To investigate and evaluate the method of PCR amplification blocking associated with fluorescent probe for the detection of pre-C region of HBV G1896A gene mutation.Methods The primers were designed based on the mutation of HBV DNA 1896 locus.The 3′end of primers was at 1896 site,and it was complemented with the base sequence of mutation template of 1896 site.The mismatching bases were separately introduced into the second and the third base of the primer by inverse counting from the 3′end for increasing the specificity of reaction.Results The PCR amplification for wild plasmid with the mutant primer showed an effectively blocking,but not showed blocking for the mutant plasmid (G1896A).The sensitivity of detection for the mutant plasmid was 5?103 copies/ml.Ninety-five cases of HBV-positive serum was selected randomly and amplified with the mutant primer,and 8 cases were positive HBV G1896A gene mutant(mutant rate of 8.4%).Conclusion The amplification blocking associated with fluorescent probe for the detection of HBV G1896A gene mutation is a effective,convenient method for the detection of clinical samples.

According to the latest research findings on nitrogen removal in both domestic and abroad, the mechanism for simultaneous nitrification and denitrifi cation(SND)was reviewed and analyzed in various aspects including macroenviron ment and microenvironment theory as well as biology theory The works such as t he realization of SND by aerobic granular sludge, simultaneous nitrogen and phos phorus removal, which could be done in the future, were proposed

Objective To observe the change of the protein and gene expression of hypoxia inducing factor-1α(HIF-1a) and vascular endothelial growth factor (VEGF) in the nude mouse tumor,which has been treated by HIFU combined with nanoscale ultrasound molecular probes with HSV1-TK gene microvascular density.Methods Sixty nude mice were implanted with HepG2 Cells to establish subcutaneous transplanted tumor.Divided this mice into six groups at random after treated by HIFU:MB+ HSV-TK+ GPC3 (group A),MB + HSV-TK (group B),HSV-TK +GPC3 (group C),HSV-TK (group D),MB + GPC3 (group E),PBS (group F).They were injected into the tail vein every after 3 days.Mice in group A,B,D and E were exposed to ultrasound by 2 W/cm2,1 MHz,5 mintues and 0.2 mL ganciclovi(GCV) was intraperitoneally injected at the first 48 hours after injection.After the treatment,immunohistoche were used to detect the microvascular density(MVD),Western blot and immunohistoche was employed to test the protein change of the VEGF and HIF-1α,Q-PCR was used to test the mRNA gene transcription of VEGF and HIF-1α in the tumor tissues.Results After 14 days,the protein expression of HIF-1α and VEGF in group A was significantly lower than that in group B,C,D,E and F (P＜0.05),the MVD level in the tumor is also like this,and the difference is statistically significant.Conclusion Anoscale ultrasound molecular probes with HSV1-TK can reduce the the level of VEGF,MVD and HIF-1α in the tumor which has been treated by HIFU,so it can inhibit tumor growth and improve the therapeutic efficacy after HIFU treatment.

Abstract: Objective　To understand the density changes and seasonal fluctuation of larvae and adults of Aedes albopictus in Hainan Province from 2017 to 2021, and to provide a scientific basis for the prevention and control of dengue fever and other vector-borne diseases in Hainan Province. Methods　The Breteau index method was used to monitor the larvae of Aedes mosquitoes, and the breeding sites of all small indoor and outdoor water containers and their larvae was checked and recorded. The adult mosquitoes were monitored by double lamination and light trap. Results　A total of 1 962 adult female Aedes albopictus mosquitoes were captured in Hainan Province from 2017 to 2021, accounting for 2.17% of the total captured mosquitoes. There were significant differences in mosquito species composition in different habitats in different years (χ2=312.258, P<0.01). In 2017, the density of mosquitoes in livestock sheds was the highest (9.71 per lamp·night), but the composition ratio of Aedes albopictus was the lowest (1.29%). In 2020, the mosquito density in hospitals was the lowest (2.22 mosquitoes per lamp·night), and in 2019, the proportion of Aedes albopictus in residential areas and hospitals was the highest (3.27% on average).The mosquito density in different habitats was the lowest in 2020. The proportion of Aedes albopictus in residential areas and livestock sheds showed an increasing trend from 2017 to 2021, while the proportion of other habitats showed significant fluctuations. The proportion of Aedes albopictus in hospitals showed an obvious decreasing trend in 2020. From 2017 to 2021, the average mosquito density was 5.97, 4.26, 3.82, 3.39 and 3.68 mosquitoes per lamp·night, respectively, showing a decreasing trend year by year except 2021. Except for 2019, the lure index also showed a decreasing trend, with an average value of 1.5/(hour·net), showing a decreasing trend year by year. From 2017 to 2021, the annual average density of Aedes albopictus in Hainan Province was 0.09 mosquitoes per lamp·night, showing obvious seasonal fluctuation. The Breteau index showed a slow decline from 2017 to 2021, and its annual peak was between June and August, which was 1-2 months later than the peak of adult mosquito density. Conclusions　The population density of Aedes albopictus in Hainan shows obvious seasonal characteristics. Mosquito control activities should be carried out according to the seasonal characteristics of mosquitoes to reduce the risk of dengue fever and other infectious diseases.

AIM To investigate the effect of recipient kidney function by CsA coadministration Ber used to induce immune tolerance in rats of allogenic cardiac transplantation. METHODS The authors established the SD to Wistar rats heterotopic cardiac transplantation model by Onos methods.Observe the cardiac allograft survival and levels of BUN and Cr in the recipients plasma. The recipients were classified into 5 groups randomly after heterotopic cardiac transplantation were performed. Group A (Wistar to Wistar)): Received placebo intraperitoneal injected for 21 days; Group B (SD to Wistar): Saline intraperitoneal injected for 21 days; Group C (SD to Wistar):CsA 2 mg?kg -1 ?d -1 intraperitoneal injected for 21 days; Group D(SD to Wistar):Ber 16 mg?kg -1 ?d -1 gastrointubation for 21 days; Group E(SD to Wistar): Ber 16 mg?kg -1 ?d -1 gastrointubation coadministration CsA 2 mg?kg -1 ?d -1 ip for 21 days. RESULTS The levels of BUN and Cr in recipint plasma is lower evidently compare with the group with CsA ip simply. CONCLUSION Ber can reduce the renal toxicity in recipients by CsA which was intraperitoneal injected (ip) over a long period time.

Objective To evaluate the therapeutic effects and to summarize the clinical experience of benign prostatic hypertrophy(BPH) with inguinal hernia in the elderly 70-89 years. Methods Clinical data of 32 patients 70-89 years old simultaneously undergone transurethral resection of prostate (TURP) and plug-mesh tension-free hernia repair from July 2000 to May 2005 were retrospectively analyzed and followed up. Results The average operating time was (85.0?12.8)minutes, the average blood loss was (90.0? 18.7 )ml. No postoperative death or life threatening complications were revealed. By 7-40 months of following up reported that there were no recurrence of hernia as well as no incontinence and urethral stenosis or other complications. IPSS,maximal flow rate and residual urine were evidently improved after operation. Conclusion Combined TURP and plug-mesh tension-free hernia repair is a safe and effective procedure for the elderly patients.

This study investigated the effects of hyperoxia on dynamic changes of thioredoxin-1 (Trx1) and thioredoxin reductase-1 (TrxR1) in alveolar type II epithelial cells (AECII) of premature rats. Pregnant Sprague-Dawley rats were sacrificed on day 19 of gestation. AECII were isolated and purified from the lungs of premature rats. When cultured to 80% confluence, in vitro cells were randomly divided into air group and hyperoxia group. Cells in the hyperoxia group were continuously exposed to 95% O(2)/5% CO(2) and those in the air group to 95% air/5% CO(2). After 12, 24 and 48 h, cells in the two groups were harvested to detect their reactive oxygen species (ROS), apoptosis, TrxR1 activity and the expressions of Trx1 and TrxR1 by corresponding protocols, respectively. The results showed that AEC II exposed to hyperoxia generated excessive ROS and the apoptosis percentage in the hyperoxia group was increased significantly at each time points as compared with that in the air group (P<0.001). Moreover, TrxR1 activity was found to be markedly depressed in the hyperoxia group in comparison to that in the air group (P<0.001). RT-PCR showed the expressions of both Trx1 and TrxR1 mRNA were significantly increased in AECII exposed to hyperoxia for 12 and 24 h (P<0.01), respectively. At 48 h, the level of Trx1 mRNA as well as that of TrxR1 mRNA in the hyperoxia group was reduced and showed no significant difference from that in the air group (P>0.05). Western blotting showed the changes of Trx1 protein expressions in the hyperoxia group paralleled those of Trx1 mRNA expressions revealed by RT-PCR. It was concluded that hyperoxia can up-regulate the protective Trx1/TrxR1 expressed by AECII in a certain period, however, also cause dysfunction of the cytoplasmic thioredoxin system by decreasing TrxR1 activity, which may contribute to the progression of oxidative stress and cell apoptosis and finally result in lung injury.

Objective:To explore the intervention effect of motivational interviews based on timing theory on self-efficacy, negative affect and coping styles of parents with infantile spasms children.Methods:Cluster sampling was used to select 82 parents of infantile spasms hospitalized in the Department of Neurology of a children’s hospital, a three-A hospital from January 2019 to October 2019. They were divided into control group and observation group with 41 cases each according to random number table. The control group received routine health education, and the observation group received five motivational interviews based on timing theory interventions on the basis of routine care. The effect of the intervention was evaluated by General Self-Efficacy Scale (GSES), Hospital Anxiety and Depression Scale (HADS), and the Chinese version of Coping Health Inventory for Parents (CHIP) before intervention, on the day of discharge, and 3 months after discharge.Results:Before the intervention, there was no significant difference in the scores of GSES, HADS and CHIP scales between the parents of the two groups ( P>0.05). After intervention, The GSES scores of the observation group on the day of discharge and 3 months after discharge were (19.63±0.87) and (22.58±1.28) points, which were significantly higher than (18.92±0.74) and (19.46±1.25) points of the control group. The difference between both groups was statistically significant ( t values were -3.865, -10.926, P<0.01). HADS-A/HADS-D scores of the observation group on the day of discharge and 3 months after discharge were (12.50±0.82), (10.50±0.87) and (9.78±0.80), (8.63±0.87) points, respectively. The HADS-A/HADS-D scores of the control group on the day of discharge and 3 months after discharge were (12.92±0.74), (11.72±0.99) and (10.23±0.78), (9.38±1.04) points, respectively. The difference was statistically significant ( t values were 2.412-5.764, P<0.05 or 0.01). The observation group scores on CHIP subscales on the day of discharge and 3 months after discharge are higher than the control group, the difference was statistically significant (t values were -7.93--2.490, P<0.05 or 0.01). Conclusions:Motivational interviews based on timing theory can enhance parents’ self-efficacy, improve their negative emotions and family coping styles, and thereby promote the recovery of children.

The present study investigated the relationship between DNA-dependent protein kinase (DNA-PK) and radiosensitivity of nasopharyngeal carcinoma (NPC) cell lines. The dose-survival relationship for NPC cell lines, CNE1 and CNE2, was analyzed using clonogenic formation assay, the activity of DNA-PK of the two cell lines was measured using the Signa TECT DNA-PK assay kit, and the localization and expression of Kus (a heterodimer) and DNA-PKcs protein in CNE1 and CNE2 before irradiation and 15 min, 1 h, 6 h, 12 h, 24 h after 4 Gy irradiation were analyzed by immunofluorescence, laser scanning confocal microscope (LSCM) and Western blot. The results showed that the surviving fraction of CNE1 was higher than that of CNE2 at each dose. The DNA-PK activity of CNE1 was also significantly higher than that of CNE2 before and after irradiation (P<0.05), while the expression of total Ku70/Ku80 in CNE1 and CNE2 had no significant difference. Increasing translocation of Ku70 and Ku80 from the cytoplasm to the nuclei in the two cell lines was observed with increase of irradiation time as detected by Western blot, and the immunofluorescence of the DNA-PK complex subunits showed greater nuclear translocation in CNE1 than CNE2 after irradiation. The results suggest that the relatively higher radio-resistance of CNE1 correlates with the higher activity of DNA-PK as compared to that of more radiosensitive CNE2 (or lower radio-resistance) before and after irradiation. Thus, DNA-PK activity may be a useful predictor of radiosensitivity of NPC.
Carcinoma ; Cell Line, Tumor ; enzymology ; radiation effects ; DNA-Activated Protein Kinase ; metabolism ; Humans ; Nasopharyngeal Neoplasms ; enzymology ; Radiation Tolerance

OBJECTIVE: To observe the correlation between the expression of galectin-7 and S100A9 with the development of cervical squamous carcinoma. METHODS: Immunohistochemical SP staining was used to detect the expression of galectin-7 and S100A9 in 243 patients with cervical intraepithelial neoplasia (CIN) or cervical squamous carcinoma. The association of clinical data with galectin-7 and S100A9 expression was examined. RESULTS: The expression of galectin-7 and S100A9 in CIN and cervical squamous carcinoma was significantly different (P<0.05). The positive rates of galectin-7 in normal cervical tissues, CIN I, CIN II, CIN III, and cervical squamous carcinoma were 56.7%, 41.9%, 32.0%, 27.3%, and 25.0%, respectively. Statistic analysis found significant difference between the normal cervical tissues and cervical squamous carcinoma (P<0.0045). The positive rates of S100A9 in CIN I, CIN II, CIN III, and cervical squamous carcinoma were 80.0%, 77.4%, 48.0%, 27.3%, and 20.2%. Statistic analysis showed significant difference between the normal tissues and CIN III, the normal cervical tissues and cervical squamous carcinoma, CIN I and CIN III, CIN I and cervical squamous carcinoma, CIN II and cervical squamous carcinoma (P<0.0045). A positive correlation was found between galectin-7 and S100A9 expression in CIN and cervical squamous carcinoma (rs=0.298, P<0.001). Expressions of both galectin-7 and S100A9 in cervical squamous carcinoma were associated with the clinical stage and lymph nodes (P<0.05), but not with patient's age and degree of differentiation (P>0.05). Expression of galectin-7 was associated with the survival rate of patients with cervical squamous carcinoma (P<0.05). Univariate analysis of Cox proportional hazards regression model revealed that the FIGO stage, lymph nodes metastasis, and the expression of galectin-7 were relevant to the 5 year survival rate of patients with cervical squamous carcinoma, which was confirmed by multiple analysis of Cox proportional hazards regression model. CONCLUSION Expression of galectin-7 and S100A9 is related with cervical the tumorigenesis of carcinoma, clinical stage, and lymph nodes of cervical squamous carcinoma. Galectin-7 is probably associated with the prognosis. The long-term survival of patients with cervical carcinoma may be associated with FIGO stage, lymph node metastasis, and the expression of galectin-7.
Calgranulin B ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Cell Differentiation ; Cervical Intraepithelial Neoplasia ; metabolism ; Female ; Galectins ; metabolism ; Humans ; Lymphatic Metastasis ; Prognosis ; Proportional Hazards Models ; Survival Rate ; Uterine Cervical Neoplasms ; metabolism