Objective To evaluate the effect of sitaxsentan on renal microcirculation in beagle dogs undergoing cardiopulmonary bypass(CPB)when ultrasound microbubble angiography was used to monitor renal microcirculation.Methods Eighteen male Beagle dogs,weighing 10-15kg,aged 2-4 yr,were allocated into 3 groups(n=6 each)using a random number table:sham operation group(Sham group),CPB group and sitaxsentan group(S group).Sitaxsentan 0.7 mg/kg was infused over 30min starting from 1 h before CPB in group S.Before CPB(T1),at 1 h of CPB(T2),at the end of CPB(T3)and at 2h after the end of CPB(T4),the time-intensity curve of renal parenchyma perfusion was obtained using ultrasound microbubble angiography,and quantitative parameters including the slope rate of ascending curve(A),area under curve(AUC),derived peak intensity(DPI)and time to peak(TTP)were fitted.Results Compared with Sham group,the value of A was significantly decreased at T2-4,AUC and TTP were increased at T3,4,DPI was decreased at T4 in renal cortex and medulla in CPB group,and the value of A was significantly decreased and TTP was increased at T2-4,AUC was increased at T3,4(P<0.05),and no significant change was found in DPI in renal cortex and medulla in S group(P>0.05).Compared with CPB group,the value of A was significantly increased and AUC and TTP were decreased at T3,4 in renal cortex and medulla(P<0.05),and no significant change was found in DPI in renal cortex and medulla in group S(P>0.05).Conclusion Sitaxsentan can improve renal microcirculation in beagle dogs undergoing CPB.

Objective To explore the mechanism of cell apoptosis of immortalized human keratinocytes (HaCaT cells) and protein expression related to this process after long term exposure to sodium arsenite (NaAsO2,1.0 μmol/L).Methods Malignant transformation model was set up through long-term exposure of HaCaT cells to 1.0 μmol/L NaAsO2.Cell passage for 0,1,7,14,21,28 and 35 generations in the process of malignant transformation were collected for measurement of cell apoptosis rate by flow cytometry,and apoptosis related proteins by Western blotting,including activation of cysteine protease 3,8 (cleaved-caspase-3,8),C/EBP homologous protein (CHOP),B-cell leukemia/lymphoma 2 (Bcl-2),and Bcl-2 associated X protein (Bax).Results Along with the arsenite treatment,the apoptosis levels were significantly decreased (F =26.770,all P ＜ 0.05),the apoptosis levels (0.307 ± 0.049,0.213 ± 0.055,0.163 ± 0.057,0.147 ± 0.035,0.053 ± 0.012) of the 7th,14th,21st,28th and 35thgenerations of cells after arsenite treatment were lower than that of control group of the 0th generation (0.393 ±0.021,all P ＜ 0.05).Compared between generations,there were statistical differences of the protein expression levels of cleaved-caspase-3,Chop,Bax and Bcl-2 in arsenite group (cleaved-caspase-3:1.000 ± 0.000,1.030 ± 0.027,1.104 ± 0.069,1.016 ± 0.087,0.838 ± 0.075,0.753 ± 0.082,0.677 ± 0.073;Chop:1.000 ± 0.000,1.059 ± 0.018,0.934 ± 0.095,0.976 ± 0.216,0.793 ± 0.136,0.651 ± 0.042,0.564 ± 0.056;Bax:1.000 ± 0.000,1.069 ± 0.037,1.028 ± 0.042,0.954 ± 0.118,0.641 ± 0.135,0.531 ± 0.132,0.429 ± 0.085;Bcl-2:1.000 ± 0.000,1.072 ± 0.023,1.249 ± 0.134,1.334 ± 0.143,1.633 ± 0.221,1.507 ± 0.152,1.461 ± 0.145,F =7.730,7.355,27.802,12.438,all P ＜ 0.05),compared with control group of the 0th generation (1.000 ± 0.000) and the same generation control group (1.000 ± 0.000),after the 21st generation,the differences were statistically significant (all P ＜ 0.05),while there was no difference of the protein expression levels of cleaved-caspase-8 (F =0.832,P ＞ 0.05).Conclusion In the process of malignant transformation,the apoptosis levels of HaCaT cells are inhibited after long term sodium arsenite exposure through mitochondria and endoplasmic reticulum (ER) stress signaling pathways.

Objective:To evaluate the biological characters of C57-TgN(HBV adr2.0)SMMU transgenic mice. Methods: Integration,expression,replication and histology change of hepatitis B virus gene in F6 transgenic mice were estimated by ge-nomic DNA PCR,Western blotting,ELISA,immunohistochemistry,serum DNA PCR,transmission electron microscopy and H-E staining. Results: Hepatitis B virus gene was integrated into F6 C57-TgN(HBV adr2. 0)SMMU transgenic mice and expressed HBsAg,HBcAg and X protein in liver tissue. HBsAg and HBeAg were expressed in serum of 19. 54% and 3. 39% F6 transgenic mice. Hepatitis B virus were replicated in serum and liver tissue of transgenic mice. Long-term integration,expression and replication of hepatitis B virus gene induced pathological lesion of transgenic mice liver and lung. Conclusion: C57-TgNCHBV adr2. 0)SMMU transgenic mice line has the biological characters including integration of hepatitis B virus gene into genomic DNA,expression and replication of hepatitis B virus gene in serum and liver, and histological change in liver and lung. It is a valuable animal system to study pathogenesis, treatment and prevention of hepatitis B virus.

Objective:To investigate the clinical efficacy of acupoint pressing acupuncture combined with moxibustion at Baihui acupoint on infantile cold and nasal obstruction caused by exogenous wind and cold. Methods:This study included a total of 120 children with exogenous wind and cold-induced infantile cold and nasal obstruction who were treated at the Jinhua Maternal & Child Health Care Hospital from February 2021 to May 2022. They were divided into three groups using a random number table method, namely the acupoint pressing acupuncture group, the moxibustion group, and the combined therapy group, each containing 40 children. Based on routine treatment, children in the acupoint pressing acupuncture group received acupoint ( Yintang, Shuangyingxiang, Shuangbitong) pressing acupuncture treatment, while those in the moxibustion group underwent mild moxibustion at Baihui acupoint, and those in the combined therapy group underwent acupoint ( Yintang, Shuangyingxiang, Shuangbitong) pressing acupuncture Baihui combined with mild moxibustion at Baihui acupoint. The clinical efficacy of each group was evaluated. At 2, 24, 48, and 72 hours after treatment, differences in nasal congestion symptom scores were compared among the three groups. Sleep quality was also compared among the three groups before and after treatment. Results:The response rate in the combined therapy group was 92.5% (37/40), which was significantly higher than 75% (30/40) in the acupoint pressing acupuncture group and 65% (26/40) in the moxibustion group ( χ2 = 4.50, 9.04, both P < 0.05). At 2, 24, 48, and 72 hours after treatment, the nasal congestion symptom score in the combined therapy group was (2.05 ± 0.55) points, (1.80 ± 0.64) points, (1.33 ± 0.59) points, and (0.90 ± 0.18) points, respectively, while it was (2.43 ± 0.59) points, (2.15 ± 0.57) points, (1.73 ± 0.84) points, and (1.18 ± 0.80) points, respectively, in the acupoint pressing acupuncture group, and (2.50 ± 0.59) points, (2.13 ± 0.78) points, (1.88 ± 0.81) points, and (1.45 ± 0.81) points, respectively, in the moxibustion group. At the above-mentioned time points, the nasal congestion symptom score was statistically significant among the three groups ( F = 3.15, 9.27, 16.17, 20.22, all P < 0.05). After treatment, daytime sleep duration and nocturnal sleep duration in the combined therapy group were (3.41 ± 0.31) hours and (12.36 ± 1.17) hours, respectively, which were significantly longer than (2.95 ± 1.07) hours and (11.33 ± 1.38) hours in the acupoint pressing acupuncture group and (2.93 ± 0.98) hours and (11.21 ± 1.93) hours in the moxibustion group ( F = 6.37, 12.31, both P < 0.05). Nocturnal wake time, the number of night wakings, and sleep onset time in the combined therapy group were (18.74 ± 2.21) minutes, (1.64 ± 0.18) times, and (15.43 ± 2.03) minutes, respectively, which were significantly shorter or less than (21.13 ± 3.78) minutes, (2.15 ± 0.66) times, and (17.63 ± 5.24) minutes in the acupoint pressing acupuncture group, and (20.53 ± 2.90) minutes, (2.11 ± 0.32) times, and (17.22 ± 2.88) minutes in the moxibustion group ( F = 15.93, 15.36, 10.11, all P < 0.05). There was a significant difference in sleep quality score among the three groups ( F = 23.45, P < 0.05). Conclusion:The combination of acupoint pressing acupuncture and moxibustion at Baihui acupoint is highly effective against infantile cold and nasal obstruction caused by exogenous wind and cold. The combined therapy can alleviate the symptoms of nasal congestion in children and improve sleep quality.

Objective: To understand the iodine nutrition status of school children aged 8-10 years in Luohu District of Shenzhen City, so as to provide basis for scientific iodine supplement for special population in the district. Methods: From 2018 to 2022, edible salt and random urine samples of non boarding students aged 8-10 years Luohu District of Shenzhen were collected for iodine content detection and follow up investigation. Meanwhile, thyroid volume was measured by ultrasonography. Results: From 2018 to 2022, a total of 1 000 salt samples were tested, of which the median salt iodine content was 26.2 mg/kg, the edible rate of iodized salt was 90.3%, and the edible rate of qualified iodized salt was 84.0%. The median urinary iodine of children was 201 μg/L. The proportion of iodine undernutrition in children who consumed qualified iodized salt was 11.0%, and the proportion of iodine undernutrition in children who consumed unqualified iodized salt was 35.0%. The goiter rate ranged from 1.0% to 2.5% between 2018 and 2022. Conclusion The rate of qualified iodine coverage and goiter of children aged 8-10 in the district reached the national standard for iodine deficiency disorders elimination, the median urinary iodine reached the appropriate level of iodine, and the iodine nutritional status was good. The iodine nutritional status of children who eat qualified iodized salt is better than that of children who eat unqualified iodized salt.

Objective: To clone mouse rod opsin promoter (ROP) and establish transgenic mice harboring mouse rod opsin promoter and enhanced green fluorescent protein(mROP-EGFP) fusion gene. Methods: Mouse ROP was cloned from C57BL/6 mouse genomic DNA by polymerase chain reaction (PCR). Expression vector of mROP-EGFP fusion gene were constructed by recombination DNA technique. It was identified by restriction endonucleases digestion and confirmed by DNA sequencing. After Not I restriction endonuclease digestion, the coding elements were microinjected into male pronuclei of mice zygotes to generate transgenic mice. The pups were evaluated by PCR at genomic DNA level and mated with normal mouse. Expression of GFP in retina of transgenic mice was detected by fluorescent microscope. Results: 2. 1 kb mouse rod opsin promoter fragment was amplified from mice genome DNA. Expression vector pmROP-EGFP was constructed successfully. Following microinjection of coding sequence of pmROP-EGFP, 3 pups were verified to integrate the mROP-EGFP fusion gene in their genomic DNA by PCR assay, named C57-TgN (mROP-EGFP )SMMU21, C57-TgN (mROP-EGFP)SM-MU26 and C57-TgN(mROP-EGFP) SMMU27. They could express GFP in retina. Conclusion: 2. 1 kb mouse rod opsin promoter is cloned and expression vector pmROP-EGFP is constructed. mROP-EGFP fusion gene transgenic mice are established, which harboring mROP-EGFP gene and expressing GFP in their retina. This is valuable for studying the development of brain and retina, pathogenesis of retina disorder and retina transplanting.

Objective:To explore the role of nuclear transcription factor erythrocyte line-2p45 (NF-E2) related factor-2 (NRF2) on autophagy during malignant transformation of immortalized human keratinocytes (HaCaT) induced by sodium arsenite (NaAsO 2). Methods:Using cell culture methods, long-term cultured HaCaT cells in DMEM high-glucose medium containing 0.0 (control group) and 1.0 μmol/L NaAsO 2 (arsenic-exposed group) to the 35th generation were used to construct a cell malignant transformation model, and 0, 1, 7, 14, 21, 28 and 35th generation cells of control group and arsenic-exposed group were collected during establishment of cell malignant transformation model. The NRF2 siRNA, phosphatidylinositol-3-hydroxykinase (PI3K) inhibitor LY294002 and mammalian target of rapamycin (mTOR) inhibitor Rapamycin were used to treat the 35th generation of malignant transformed HaCaT cells in arsenic-exposed group (T-HaCaT). The protein expressions of NRF2, PI3K-protein kinase B (Akt)-mTOR signaling pathway related indicators PI3K, Akt, mTOR, phosphorylated (p)-PI3K, p-Akt, p-mTOR, autophagy-related proteins p62, Beclin1, microtubule-associated protein-1 light chain (LC)3Ⅰ, and LC3Ⅱof different generations HaCaT cells in control group and arsenic-exposed group, and T-HaCaT cells of each treatment group were determined by Western blotting. Results:There were significant differences in the NRF2 protein and the ratios of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR between different generations HaCaT cells in arsenic-exposed group ( F = 9.371, 16.035, 15.932, 27.739, P < 0.05), and they were higher than NRF2 protein and ratio of p-mTOR/ mTOR of the same generation in control group ( P < 0.05). Compared with HaCaT cells of the same generation, the expressions of NRF2, p-PI3K, p-Akt, p-mTOR and p62 proteins in T-HaCaT cells were significantly higher, Beclin1 protein expression and the ratio of LC3Ⅱ/LC3Ⅰ were significantly lower ( P < 0.05). The NRF2 silenced T-HaCaT cells had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expressions of NRF2, p-mTOR and p62 than the corresponding control siRNA (Con siRNA) group ( P < 0.05). The T-HaCaT cells in LY294002 treatment group had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expressions of NRF2, p-PI3K, p-Akt and p-mTOR proteins than the corresponding non-treatment group ( P < 0.05). The T-HaCaT cells in Rapamycin treatment group had higher expression of Beclin1 and the ratio of LC3Ⅱ/LC3Ⅰ, and lower expression of p-mTOR protein than the corresponding non-treatment group ( P < 0.05). Conclusions:During the arsenic-induced malignant transformation of HaCaT cells, NRF2 can act as a downstream factor of PI3K-Akt and an upstream factor of mTOR in PI3K-Akt-mTOR signaling pathway, an important regulatory mechanism of autophagy. This abnormal expression of autophagy may eventually lead to malignant transformation of cells.

At the present day, curettage and periodontal surgery comprise the main strategy for the treatment of periodontitis, however, these methods are limited in regenerating cementum. It has been found that some biological factors such asenamel matrix derivative (EMD), transforming growth factor-β (TGF-β) and insulin-like growth factor (IGF) could promote cementum regeneration. In the cementum regenerationstudies, there has been a lack of criteria to distinguish cementum from alveolar bone and other types of cementum. Therefore, this article will briefly review the biological factors that affect the cementum regeneration and the molecular markers used to judge the regenerating cementum.

Objective: To investigate the effect of triptolide, a specific inhibitor of heat shock protein 70 (HSP70), on apatinib resistance in gastric cancer cells line MKN45. Methods: The apatinib-resistant cells (MKN45/AR) and MKN45 parental cells were treated with apatinib, triptolide and apatinib combined with triptolide, respectively. CCK-8 assay was performed to determine the half maximal inhibitory concentration (IC₅₀) of MKN45/AR and MKN45 cells in the presence of different treatment. The mRNA expression of heat shock protein gene (HSPA1A and HSPA1B) was detected by RT-PCR, while the protein expression of heat shock protein 70 was analyzed using Western blot in MKN45/AR and MKN45 cells. Results: The IC₅₀ values of apatinib-sensitive and apatinib-resistant MKN45 cells were 10.411 μmol/L and 70.527 μmol/L, respectively, showing a significant difference (P<0.05). The mRNA expression of HSPA1A and HSPA1B in MKN45/AR cells was significantly higher than that in MKN45 cells (P<0.001). The protein expression of heat shock protein 70 was significantly decreased after 0.25 μmol/L triptolide treatment in MKN45/AR cells (P<0.01). When heat shock protein 70 was inhibited by triptolide, the IC₅₀ value of apatinib in MKN45/AR cells was reduced to 11.679 μmol/L, which was significantly lower than cells treated with apatinib alone (P<0.05). Conclusions The apatinib-resistant MKN45 cells have high levels of heat shock protein 70. Low doses of triptolide can significantly inhibit heat shock protein 70, leading to reverse the resistance phenotype of MKN45/AR cells. Therefore, inhibition of heat shock protein 70 provides a new therapy strategy for patients with apatinib resistance.