A segmentation model to detect cevical lesions based on machine learning of colposcopic images.

Background: Semantic segmentation is crucial in medical image diagnosis. Traditional deep convolutional neural networks excel in image classification and object detection but fall short in segmentation tasks. Enhancing the accuracy and efficiency of detecting high-level cervical lesions and invasive cancer poses a primary challenge in segmentation model development. Methods: Between 2018 and 2022, we retrospectively studied a total of 777 patients, comprising 339 patients with high-level cervical lesions and 313 patients with microinvasive or invasive cervical cancer. Overall, 1554 colposcopic images were put into the DeepLabv3+ model for learning. Accuracy, Precision, Specificity, and mIoU were employed to evaluate the performance of the model in the prediction of cervical high-level lesions and cancer. Results: Experiments showed that our segmentation model had better diagnosis efficiency than colposcopic experts and other artificial intelligence models, and reached Accuracy of 93.29 %, Precision of 87.2 %, Specificity of 90.1 %, and mIoU of 80.27 %, respectively. Conclution: The DeepLabv3+ model had good performance in the segmentation of cervical lesions in colposcopic post-acetic-acid images and can better assist colposcopists in improving the diagnosis.

First Report of Tomato spotted wilt virus infecting Helichrysum bracteatum in China.

Strawflower (Helichrysum bracteatum, Asteraceae) , an annual or biennial herb, is one of the most popular flowers in the world because of the colorful flowers and the long flowering period. However, the ornamental plants belonging to Asteraceae are susceptible to numerous viruses such as cucumber mosaic virus (CMV) (Cucumovirus, Bromoviridae) , potato virus Y (Potyvirus, Potyviridae), tomato mosaic virus (ToMV) (Tobamovirus, Virgaviridae), tobacco mosaic virus (TMV) (Tobamovirus, Virgaviridae), chrysanthemum virus B (CVB) (Carlavirus, Betaflexiviridae), tomato aspermy virus (TAV) (Cucumovirus, Bromoviridae), tomato spotted wilt virus (TSWV) (Orthotospovirus tomatomaculae, Tospoviridae), and impatiens necrotic spot virus (INSV) (Orthotospovirus impatiensnecromaculae, Orthotospovirus) resulting in severe yield loss (Verma et al. 2003; Raj et al. 2007; Kondo et al. 2011; Liu et al. 2014; Marys et al. 2014; Min et al. 2020; Gautam et al. 2021; Read et al. 2022; Supakitthanakorn et al. 2022). Among these viruses, the TSWV, a thrips-transmitted negative-stranded RNA virus, is well known to cause viral disease in several plant species while is less reported in Helichrysum, especially in China. In April 2021, viral attack symptoms were detected on the leaves of H. bracteatum during our routine checks in the greenhouse located at Shunyi District, Beijing, China, such as wilting, shrinking, chlorotic blotches, chlorotic ring spots. To investigate the virus infecting H. bracteatum, in total of 25 symptomatic and 5 asymptomatic leaves were sampled and tested by the effective double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using antisera against CMV, PVY/PVX, ToMV, TMV, CVB, TAV, TSWV, INSV, separately (Agdia, USA). Only the TSWV showed positive in symptomatic samples, and asymptomatic samples were all negative, which implied TSWV infection. To further confirm the virus type of TSWV isolated from H. bracteatum samples, the genomic RNA of the virus was isolated using reverse transcription and polymerase chain reaction (RT-PCR), and then was cloned, sequenced and analyzed. Total RNA of five symptomatic leaves (ELISA-positive) were extracted using the FastPure Plant Total RNA Isolation Kit (Vazyme, China), and then were reverse transcribed by HiScript II Reverse Transcriptase (Vazyme, China). Each genome segments were amplified using Phanta Max Super-Fidelity DNA Polymerase (Vazyme, China) with TSWV-specific primers newly designed and listed in Table S1. The PCR setup was as follow: 95°C for 30 s, followed by 35 cycles at 95°C for 30 s, 55°C for 30 s, and 72°C for 1.5 min, with a final extension at 72°C for 10 min. All PCR products were cloned into the TA/Blunt-Zero vector (Vazyme, China) and sequenced (GENEWIZ, Inc.). We assembled and then analyzed the evolutionary relationship of three genomic fragments, that is, TSWV-BJFC-Hb S (2923 bp), M (4785 bp) and L (8971 bp) using the BLAST tools. The results showed high similarity with TSWV-henan isolated from pepper in China (99.6% to TSWV-S (MT799179.1), 99.8 % to TSWV-M (MT799178.1) and 99.8 % TSWV-L (MT799177.1)). These sequences have been submitted to the GenBank (OM982910, OM982911 and OM937131). Taking all of these evidences together, the viral disease observed in H. bracteatum was closely associated with TSWV. TSWV is currently widespread in China, infecting Nasturtium, Chrysanthemum and cowpea (Xiao et al. 2015; Hu et al. 2018; Yu et al. 2021). Epidemics of TSWV have also been reported in several other countries such as Korea, North Carolina, Turkey and India (Renukadevi et al. 2015; Koehler et al. 2016; Kwak et al. 2021; Erilmez, S. 2022). This is the first report of TSWV infection on H. bracteatum in China. Due to the fast spread and serious economic losses of TSWV, the rapid detection may be the essential way to prevent this viral disease among crops (Macharia et al. 2014).

Kuwanon G protects HT22 cells from advanced glycation end product-induced damage.

The incidence of diabetic encephalopathy is increasing as the population ages. Evidence suggests that formation and accumulation of advanced glycation end products (AGEs) plays a pivotal role in disease progression, but limited research has been carried out in this area. A previous study demonstrated that Kuwanon G (KWG) had significant anti-oxidative stress and anti-inflammatory properties. As AGEs are oxidative products and inflammation is involved in their generation it is hypothesized that KWG may have effects against AGE-induced neuronal damage. In the present study, mouse hippocampal neuronal cell line HT22 was used. KWG was shown to significantly inhibit AGE-induced cell apoptosis in comparison with a control treatment, as determined by both MTT and flow cytometry. Compared with the AGEs group, expression of pro-apoptotic protein Bax was reduced and expression of anti-apoptotic protein Bcl-2 was increased in the AGEs + KWG group. Both intracellular and extracellular levels of acetylcholine and choline acetyltransferase were significantly elevated after KWG administration in comparison with controls whilethe level of acetylcholinesterase decreased. These changes in protein expression were accompanied by increased levels of superoxide dismutase and glutathione peroxidase synthesis and reduced production of malondialdehyde and reactive oxygen species. Intracellular signaling pathway protein levels were determined by western blot and immunocytochemistry. KWG administration was found to prevent AGE-induced changes to the phosphorylation levels of Akt, IκB-α, glycogen synthase kinase 3 (GSK3)-α and ß, p38 MAPK and NF-κB p65 suggesting a potential neuroprotective effect of KWG against AGE-induced damage was via the PI3K/Akt/GSK3αß signaling pathway. The findings of the present study suggest that KWG may be a potential treatment for diabetic encephalopathy.

Neuraxial adjuvants for prevention of perioperative shivering during cesarean section: A network meta-analysis following the PRISMA guidelines.

BACKGROUND: Perioperative shivering is clinically common during cesarean sections under neuraxial anesthesia, and several neuraxial adjuvants are reported to have preventive effects on it. However, the results of current studies are controversial and the effects of these neuraxial adjuvants remain unclear. AIM: To evaluate the effects of neuraxial adjuvants on perioperative shivering during cesarean sections, thus providing an optimal choice for clinical application. METHODS: A systematic review and network meta-analysis were conducted following the PRISMA (Preferred Reported Items for Systematic Review and Meta-analysis) guidelines. Analyses were performed using Review Manager 5.3 and Stata 14.0. We searched PubMed, EMBASE, Web of Science, and Cochrane Central databases for eligible clinical trials assessing the effects of neuraxial adjuvants on perioperative shivering and other adverse events during cesarean sections. Perioperative shivering was defined as the primary endpoint, and nausea, vomiting, pruritus, hypotension, and bradycardia were the secondary outcomes. RESULTS: Twenty-six studies using 9 neuraxial adjuvants for obstetric anesthesia during caesarean sections were included. The results showed that, compared with placebo, pethidine, fentanyl, dexmedetomidine, and sufentanil significantly reduced the incidence of perioperative shivering. Among the four neuraxial adjuvants, pethidine was the most effective one for shivering prevention (OR = 0.15, 95%CI: 0.07-0.35, surface under the cumulative ranking curve 83.9), but with a high incidence of nausea (OR = 3.15, 95%CI: 1.04-9.57) and vomiting (OR = 3.71, 95%CI: 1.81-7.58). The efficacy of fentanyl for shivering prevention was slightly inferior to pethidine (OR = 0.20, 95%CI: 0.09-0.43), however, it significantly decreased the incidence of nausea (OR = 0.34, 95%CI: 0.15-0.79) and vomiting (OR = 0.25, 95%CI: 0.11-0.56). In addition, compared with sufentanil, fentanyl showed no impact on haemodynamic stability and the incidence of pruritus. CONCLUSION: Pethidine, fentanyl, dexmedetomidine, and sufentanil appear to be effective for preventing perioperative shivering in puerperae undergoing cesarean sections. Considering the risk-benefit profiles of the included neuraxial adjuvants, fentanyl is probably the optimal choice.

Effects of the health belief model following acute exacerbation of chronic obstructive pulmonary disease in a hospital in China.

BACKGROUND: This study aims to investigate the effects of education with health belief model (HBM) on anxiety and fatigue among patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: Patients with AECOPD admitted into Taizhou People's Hospital, Jiangsu, China between December 2012 and October 2013 were randomly selected for the present study according to random number table. A total of 99 cases with anxiety were included. These patients were divided into two groups: experimental group educated by HBM (n=47), and control group educated by conventional method (n=52). The scores for anxiety and fatigue were evaluated using the self-rating anxiety scale (SAS) and multidimensional fatigue inventory (MFI-20). RESULTS: After educational intervention by HBM, patients achieved a significant decrease in anxiety and fatigue scores, when compared to patients in the control group, who were educated by the conventional method at the time of admission, discharge, and 6 weeks after discharge (P<0.05). CONCLUSIONS: HBM program effectively may alleviate anxiety and fatigue, providing necessary information for symptom management among patients with AECOPD.

Comparison of dominant hand to non-dominant hand in conduction of reaching task from 3D kinematic data: Trade-off between successful rate and movement efficiency.

This study aimed to investigate the effects of handedness on motion accuracies and to compare 3D kinematic data in reaching performance of dominant and non-dominant hand with the influence of movement speed and target locations. Twelve healthy young adults used self-selected and fast speed to reach for three different target locations as follows: frontal, ipsilateral and contralateral to the performing hand, with equal distance. Both hands were tested and kinematic parameters were recorded by 3D motion analysis system. Successful rate, reach path ratio, mean and peak velocity, the timing of peak velocity and ROM of joints were analyzed. Reach path ratio was smaller when using the dominant hand (p < 0.01) and fast speed (p < 0.01) to perform the movement, but the successful rate of the dominant hand was lower than non-dominant hand during fast speed reaching (99.1% vs 100%). Contralateral movement had lower velocity than the other two target locations, while velocity did not vary between non-dominant and dominant hand. The timing of peak velocity occurred significantly later for fast speed movements (p < 0.01). Trunk rotation was significantly smaller when using the dominant hand, fast movement speed or reaching to the ipsilateral target. The ROM of elbow and wrist flexion-extension decreased in contralateral reaching. The performance of the dominant hand and/or fast speed movements was more efficient with straighter hand path and less trunk rotation, but the successful rate decreased in dominant hand during fast speed movements. The timing of peak velocity occurred later during fast movement in both hands indicating a decreased feedback phase. Target location can influence movement strategy as reaching to contralateral target required more proximal movements and ipsilateral reaching used more distal segment movements.

CONSORT-Assistive technology-180° rotating eating spoon improves the ability of eating of self-care patients with upper extremity dyskinesia: Rotating eating spoon improves eating ability.

OBJECTIVE: This study aims to develop an assistive technology-180° rotating feeding spoon that could improve the ability of eating of self-care patients with upper extremity dyskinesia. METHODS: The Brunnstrom 6-stage rating of hemiplegia was adopted. During the different recovery stages of the upper limbs, the patients orally ate using a feeding spoon with a non-rotatory head and a 180° rotating feeding spoon. The ability of these patients to eat by themselves was observed, and the basic activity of daily living (BADL) was assessed using the Barthel index (BI). RESULTS: The Brunnstrom assessment scale was used to analyze the results of the patient's upper limb function examination, and the results revealed that the 180° rotating feeding spoon could assist patients with different degrees of upper limb dysfunction when eating independently. CONCLUSIONS: The 180° rotating feeding spoon can assist patients with upper limb dysfunction when eating independently. For patients with different degrees of upper limb dysfunction, the spoon can provide different degrees of aid.

Parasitism and venom of ectoparasitoid Scleroderma guani impairs host cellular immunity.

Venom is a prominently maternal virulent factor utilized by parasitoids to overcome hosts immune defense. With respect to roles of this toxic mixture involved in manipulating hosts immunity, great interest has been mostly restricted to Ichneumonoidea parasitoids associated with polydnavirus (PDV), of which venom is usually considered as a helper component to enhance the role of PDV, and limited Chalcidoidea species. In contrast, little information is available in other parasitoids, especially ectoparasitic species not carrying PDV. The ectoparasitoid Scleroderma guani injects venom into its host, Tenebrio molitor, implying its venom was involved in suppression of hosts immune response for successful parasitism. Thus, we investigated the effects of parasitism and venom of this parasitoid on counteracting the cellular immunity of its host by examining changes of hemocyte counts, and hemocyte spreading and encapsulation ability. Total hemocyte counts were elevated in parasitized and venom-injected pupae. The spreading behavior of both granulocytes and plasmatocytes was impaired by parasitization and venom. High concentration of venom led to more severely increased hemocyte counts and suppression of hemocyte spreading. The ability of hemocyte encapsulation was inhibited by venom in vitro. In addition to immediate effects observed, venom showed persistent interference in hosts cellular immunity. These results indicate that venom alone from S. guani plays a pivotal role in blocking hosts cellular immune response, serving as a regulator that guarantees the successful development of its progenies. The findings provide a foundation for further investigation of the underlying mechanisms in immune inhibitory action of S. guani venom.

[Application and effect of enhanced recovery after surgery in early rehabilitation of oral cancer patients after operation].

PURPOSE: To explore the application and effect of enhanced recovery after surgery (ERAS) in early rehabilitation of oral cancer patients after operation. METHODS: Ninety cases of oral cancer admitted from January 2016 to December 2017 were randomly divided into two groups. 45 cases of oral cancer in the experimental group received ERAS nursing after operation, and another 45 patients received routine nursing after operation. The sleep duration, pain and halitosis 3 days after operation; the first time to get out of bed, the length of hospitalization; and the level of albumin and other laboratory indicators 1 and 3 days after operation were observed. SPSS 17.0 software package was used for statistical analysis. RESULTS: After 1 day and 3 day of ERAS intervention, the nutritional index of the experimental group was lower than that of the control group. The time of first ambulation and the length of hospitalization were shorter than those of the control group. The average sleep time 3 days after operation was longer than that of the control group. The incidence of halitosis, wound infection and moderate pain was lower than that of the control group. The difference was statistically significant (P<0.05). CONCLUSIONS: ERAS nursing after oral cancer surgery can improve the comfort of patients, enhance the clinical outcome, promote rapid recovery of patients, with good clinical application prospects.

Long non-coding RNA SOX21-AS1 sponges miR-145 to promote the tumorigenesis of colorectal cancer by targeting MYO6.

Emerging evidences have proved that long non-coding RNAs (lncRNAs) act as important molecular regulator in the tumor progression, including colorectal cancer (CRC). LncRNA SOX21-AS1 has been verified as oncogenic molecular in other cancers and tumorigenesis. In present study, our team investigates the clinical characteristic and molecular function in CRC carcinogenesis. Results showed that lncRNA SOX21-AS1 expression was significantly over-expressed in CRC tissue samples and cells. The aberrant over-expression of SOX21-AS1 indicated poor prognosis of CRC patients. In vitro and in vivo validation experiments, SOX21-AS1 silencing inhibited the proliferation, invasion, and decreased the tumor growth of CRC cells. Moreover, miR-145 was proved to be the target of SOX21-AS1, besides, myosin VI (MYO6) was found to be one of the targets of miR-145 using bioinformatics prediction programs and rescue confirmation experiments. In summary, our study reveals the tumorigenic effect of lncRNA SOX21-AS1 in CRC cells via targeting miR-145/MYO6, providing a novel insight for CRC carcinogenesis.

Effect of Xing Nao Kai Qiao Needling on Cognitive and Motor Function of Patients in Acute Stage of Cerebral Infarction / 上海针灸杂志

Objective To observe the effect ofXing Nao Kai Qiao (awakening brain and opening orifice) needling method on the cognitive and motor function of patients in acute stage of cerebral infarction.Method Sixty-four patients in acute stage of cerebra infarction were recruited and randomized into a trial group and a control group, 32 cases in each group. The control group was intervened by Edaravone 30 mg plus Fibrinogenase injection 200 U mixed in 500 mL 0.9% NaCl via intravenous drip, once a day. The trial group was byXing Nao Kai Qiao needling method in addition to the intervention given to the control group, once a day, 5 times a week. The two groups were both given 2-week successive treatment. At the end of the intervention, clinical efficacy, cognitive function, motor function, nervous function, serum C-reactive protein and NO levels, and adverse reactions of the two groups were compared.Result After the treatment, the Chinese Stroke Scale (CSS) score dropped significantly, Loewenstein Occupational Therapy Cognitive Assessment (LOTCA) score, Barthel Index (BI) score and motor function score of both upper and lower limbs increased significantly in both groups (P<0.05); the total effective rate of the trial group was significantly higher than that of the control group (P<0.05); the CSS score was lower, and LOTCA score, BI score, and motor function scores of both upper and lower limbs were higher in the trial group compared with those in the control group, and the differences were statistically significant (P<0.05); on the 1st, 3rd, 7th and 14th day after the intervention, the serum CRP and NO levels were significantly lower in the trial group compared with those in the control group (P<0.05). There was no significant difference in comparing the adverse reaction incidence between the two groups (P>0.05).Conclusion Xing Nao Kai Qiao needling method can produce a significant efficacy in treating patients in acute stage of cerebral infarction, and it can improve the cognitive, motor and nervous functions with high security.

Identification and feature analysis of Stephanoascus ciferrii isolated from 12 cases with chronic suppurative otitis media / 临床检验杂志

Objective To study the laboratory features of Stephanoascus ciferrii isolated from middle ear excretion of the patients with chronic suppurative otitis media for providing the basis of diagnosis for the rare fungus infection.Methods The study was performed for 12 patients who were suffered from chronic suppurative otitis media associated with Stephanoascus ciferrii.The secretions of middle ear were cultured and the macroscopic features of colony and cells of the fungi were observed.The species of the fungi was identified by Vitek 2 Compact system.Results Stephanoascus ciferrii may grow on common media such as blood plate,chocolate plate and sabourauad plate,at 28 to 35 ℃.The form of the colonies displayed cream-colored,rough,hard and wrinkled quality after 1 week incubation.On CHROMagar medium the colonies looked like small mushroom with blue center and white edge.Under microscopy,abundant branched septate mycelium with small ramified chains of oval blastoconidia and the conidia were arranged along the side of hyphae.The fungi strain may assimilate lots of carbohydrates and common amino acid.Vitek 2 Compact automated system should be able to identify Stephanoascus ciferrii.Conclusion Stephanoascus ciferrii is a rare opportunistic pathogenic fungus.In clinical laboratories special attentions should be paid to culture and identification of the fungus from middle ear excretion.

Effect of Xing Nao Kai Qiao Needling on Cognitive and Motor Function of Patients in Acute Stage of Cerebral Infarction / 上海针灸杂志

Objective To observe the effect ofXing Nao Kai Qiao (awakening brain and opening orifice) needling method on the cognitive and motor function of patients in acute stage of cerebral infarction.Method Sixty-four patients in acute stage of cerebra infarction were recruited and randomized into a trial group and a control group, 32 cases in each group. The control group was intervened by Edaravone 30 mg plus Fibrinogenase injection 200 U mixed in 500 mL 0.9% NaCl via intravenous drip, once a day. The trial group was byXing Nao Kai Qiao needling method in addition to the intervention given to the control group, once a day, 5 times a week. The two groups were both given 2-week successive treatment. At the end of the intervention, clinical efficacy, cognitive function, motor function, nervous function, serum C-reactive protein and NO levels, and adverse reactions of the two groups were compared.Result After the treatment, the Chinese Stroke Scale (CSS) score dropped significantly, Loewenstein Occupational Therapy Cognitive Assessment (LOTCA) score, Barthel Index (BI) score and motor function score of both upper and lower limbs increased significantly in both groups (P<0.05); the total effective rate of the trial group was significantly higher than that of the control group (P<0.05); the CSS score was lower, and LOTCA score, BI score, and motor function scores of both upper and lower limbs were higher in the trial group compared with those in the control group, and the differences were statistically significant (P<0.05); on the 1st, 3rd, 7th and 14th day after the intervention, the serum CRP and NO levels were significantly lower in the trial group compared with those in the control group (P<0.05). There was no significant difference in comparing the adverse reaction incidence between the two groups (P>0.05).Conclusion Xing Nao Kai Qiao needling method can produce a significant efficacy in treating patients in acute stage of cerebral infarction, and it can improve the cognitive, motor and nervous functions with high security.

VEGF shRNA enhances the sensitivity of multidrug-resistant leukemia cells to anticancer agent / 中国实验血液学杂志

This study was aimed to explore the effect of vascular endothelial growth factor (VEGF) on sensitivity of leukemia cell line K562/A02 to doxorubicin by using RNA interference, and to investigate its mechanism. The 3 shRNA targeting human vegf gene were synthesized, then transfected into K562/A02 cells by lipofectamine 2000 reagent. RT-PCR was used to detect the expression of vegf and mrp1 at the mRNA level;Western blot was used to analyze the expression of VEGF, MRP1, AKT, P-AKT at the protein level; MTT was used to determine the IC(50) value of transfected cells to doxorubicin (DOX); flow cytometry was used to detect cell apoptosis and intracellular Rho123 retention. The results showed that after vegf shRNA were transfected into K562/A02 cells, the expression of vegf at the mRNA level decreased, and the difference between vegf shRNA2 group or vegf shRNA3 group and HK group was statistically significant (p < 0.05), the greatest decrease was observed in the cells transfected with vegf shRNA3; and the protein level of VEGF was also down-regulated. The IC(50) value of positively transfected group was lower than that of control groups, and the difference between vegf shRNA2 group or vegf shRNA3 group and HK group was significant (p < 0.05). The retention of intracellular Rho123 was enhanced in three positively transfected groups (p < 0.05). Cell apoptosis increased in positively transfected groups, and there was statistically difference between vegf shRNA2 group or vegf shRNA3 group and HK group (p < 0.05). The expression of mrp1 at the mRNA level were decreased, and there were statistical difference between vegf shRNA3 group and HK group (p < 0.05), and the protein level of mrp1 was also down-regulated; the expression of P-AKT at protein level decreased in positively transfected groups, and the greatest decrease was seen in vegf shRNA3 group. It is concluded that the transfection with exogenous vegf shRNA can inhibit the expression of vegf at both mRNA and protein levels, and enhance the sensitivity of K562/A02 cell to doxorubicin, the mechanism of which may be the inhibition of apoptosis and down-regulation of MRP1 by inactivating PI3K/AKT signaling pathway.

Influence of YB-1 protein on the biological behaviour in K562/A02 cells / 中国实验血液学杂志

The aim of this study was to investigate whether the growth, apoptosis and sensitivity to anticancer agent could be altered after introduction of YB-1 shRNA eukaryotic expression vector into the K562/A02 cells, and its possible molecular mechanisms. The recombinant eukaryotic expression plasmids including YB-1 shRNA and the vector-random-sequence were introduced into K562/A02 cells by lipofectamine mediation, and the positive clones were screened by G418. RT-PCR and Western blot were employed to detect the expression of mRNA and protein of YB-1 in leukemia cells, respectively. The proliferative ability of the cells was determined by MTT assay and cell cycle analysis. Apoptosis of K562/A02 cells was assayed by AnnexinV-FITC/PI double labeled flow cytometry. The drug sensitivity to anticancer agent was determined by MTT assay. The expressions of MDR1 gene and P-gp were detected by RT-PCR and flow cytometry respectively. The results indicated that the levels of mRNA and protein of YB-1 decreased dramatically in three groups of positively transfected cells when compared with control cells. The inhibitory rates of 3 different shRNA sequences targeting YB-1 gene were (65.1 ± 2.1)%, (27.4 ± 1.3)% and (67.4 ± 1.6)% respectively. The introduction of exogenous YB-1 shRNA gene into K562/A02 cells resulted in decreased levels of the proliferative ability in K562/A02 cells, and displayed higher at G(1), lower at G(2) and S phase in cell cycle distribution in comparison with the control groups. AnnexinV/PI detection indicated higher AnnexinV(+) ratio in 3 groups of positively transfected cells after being treated with As(2)O(3) of 0.5 µmol/L for 24 hours. The IC(50) values of doxorubicin in 3 groups of positively transfected cells were significantly lower than that in control group. The level of MDR1 gene and P-gp decreased significantly in 3 groups of positively transfected cells. It is concluded that the transfection with YB-1 shRNA gene can inhibit the proliferation of leukemia cells and induce cell apoptosis. The expression of MDR1 mRNA and P-gp decrease after transfection of YB-1 shRNA into K562/A02 cells.

CD44 variant increases the invasive ability of human breast cancer cell line MCF-7 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the impact of a new CD44 variant on invasion of human breast cancer cell line MCF-7, and its possible mechanisms.</p><p><b>METHODS</b>The full length cDNA encoding CD44v17 was obtained from the total RNA isolated from the MCF-7/ADR cells by reverse transcript-polymerase chain reaction (RT-PCR) and subcloned into pMD19-T vector. The CD44v17 gene sequence and reading frame were confirmed by two restriction enzymes and nucleotide sequencing, and then inserted into the eukaryotic expression vector pcDNA3.1. The pcDNA3.1-CD44v17 was transfected into MCF-7 cells by Lipofectamine. The changes of MMP-2 and MMP-9 expression at gene and protein levels were detected by RT-PCR and gelatin zymography, respectively. The number of the cells through the artificial matrix membrane in every group was counted to compare the change of the invasive ability regulated by CD44 variant. The ERK and p-ERK were investigated by Western blotting to approach the molecular mechanisms of MMP-2 and MMP-9 expression regulated by CD44 variant.</p><p><b>RESULTS</b>The new gene sequence was successfully cloned into recombinant vector pcDNA3.1 and identified by the two restriction enzymes. It was confirmed that the reconstructed plasmid contained the sequence of CD44 gene variant which was composed of 1 to 4 exons, 16 to 17 exons, and 1 to 205 bases of 18 exons. The new gene sequence was sent to NCBI for publication and obtained the registered number FJ216964. The up-regulated levels of the CD44 gene mRNA and protein were respectively detected by RT-PCR and flow cytometry in MCF-7 cells transfected with pcDNA3.1-CD44v17. The invasiveness of the cells and the activity of MMP-2 and MMP-9 were clearly activated by hyaluronic acid (HA) treatment and blocked by CD44 neutralizing antibody. Pretreated MCF-7/CD44v17 cells with the neutralizing antibody against CD44 and the inhibitor of MAPKs signaling pathway strongly block the expression of p-ERK.</p><p><b>CONCLUSION</b>A new CD44 gene variant has been found in adriamycin-resistant human breast cancer MCF-7/ADR cells. The expression vector pcDNA3.1-CD44v17 has been cloned and constructed successfully. HA can be integrated with CD44 variant and then regulates the expression of MMP-2 and MMP-9, which increases the invasion ability of MCF-7 cells through the Ras/MAPK signaling pathway.</p>

Effect of YB-1 gene knockdown on human leukemia cell line K562/A02 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the potential effects of YB-1 gene knockdown on gene expression profile, cell growth and apoptosis in leukemia cell line K562/A02.</p><p><b>METHODS</b>The recombinant eukaryotic expression plasmid containing YB-1 short hairpin RNA (shRNA) or random-sequence (HK) were transfected into K562/A02 cells by lipofectamine mediation. cDNA microarray was performed to explore the alteration of gene expression profile when YB-1 gene expression was decreased. Expression of CARD8 and RHOC genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). The proliferative ability of the cells was determined by methyl thiazolyltetrazolium (MTT) assay and cell cycle analysis. Cell apoptosis was assayed by Annexin V-FITC/PI double labeled flow cytometry.</p><p><b>RESULTS</b>The levels of YB-1 mRNA and protein decreased dramatically in three positively transfected cells when compared with untransfected K562/A02 cells or K562/A02-HK thansfected cells. Gene expression profile was altered by transfection of YB-1 shRNA into K562/A02 cells. Among 47,000 genes on the microarray, 252 genes were detected to have changes, with 143 down-regulated and 109 up-regulated. They were functionally related to cell cycle progression, gene replication, metabolism, cell apoptosis, cell signal transduction, etc. An increase in CARD8 gene expression and a decrease in RHOC gene expression have been confirmed by RT-PCR in K562/A02-YBX13 cells. The introduction of exogenous YB-1 shRNA gene into K562/A02 cells resulted in decreased proliferation, higher G1, lower G2 and S ratio in cell cycle distribution in comparison with the control groups. Annexin V/PI detection indicated higher Annexin V+ ratio in the three positively transfected cells 24 hours after cells were treated with 0.5 micromol/L of As2O3.</p><p><b>CONCLUSION</b>Down-regulation of YB-1 gene by shRNA-YB-1 can alter the gene expression profile in K562/A02 cells, leading to change of cell proliferation and apoptosis.</p>

[Study on the mechanism of intrauterine infection of hepatitis B virus].

OBJECTIVE: To study the possible mechanism of intrauterine infection of hepatitis B virus (HBV). METHODS: HBV DNA was examined in amniotic fluid, and vaginal secretion of 59 HBsAg positive mothers and in cord blood of their neonates by PCR. Ten negative hepatitis B virus marker (HBVM) mothers and their neonates were served as control. HBsAg and HBcAg in placenta were examined by avidin biotin complex (ABC) method. RESULTS: The detection rate of HBV DNA in amniotic fluid, vaginal secretion and neonatal cord blood of the study group were 47.5% (28/59), 52.5% (31/59) and 45.8% (27/59) respectively. HBsAg and HBcAg in placenta was distributed in the following descending order: maternal decidual cells, trophoblastic cells, villous mesenchymal cells and villous capillary endothelial cells. But the distribution was in reverse order in 4 placentas. HBsAg and HBcAg were detected in amniotic epithelial cells in 32 mothers. CONCLUSION: The main route of HBV transmission from mother to fetus is transplacental, from maternal side of placenta to fetal side. However, HBV intrauterine infection may take place through other routes.

Studies on intrinsic ionization constants of Fe-Al-Mg hydrotalcite-like compounds.

The theoretical analysis of the intrinsic ionization constant (K(a2)(int)) of Fe-Al-Mg hydrotalcite-like compounds (HTlc) possessing permanent charges was first performed using the double extrapolation method proposed by James et al. The theoretical permanent charge density (sigma(p,T)) of the HTlc sample was calculated from the crystal structure of HTlc, and the influence of sigma(p,T) on the K(a2)(int) was also examined. From the experimental results, these conclusions can be obtained: the zero point of the charge (pH(ZPC)) of Fe-Al-Mg HTlc increases with decreased Fe3+ content and increases with increased Mg2+ in the HTlc. The pK(a2)(int) of Fe-Al-Mg HTlc also increases with decreased Fe3+ and increased Mg2+ content in the sample; furthermore, the pK(a2)(int) of Fe-Al-Mg HTlc increases with decreased sigma(p,T).