Altered gut microbiota composition in children and their caregivers infected with the SARS-CoV-2 Omicron variant.

BACKGROUND: Gut microbiota alterations have been implicated in the pathogenesis of coronavirus disease 2019 (COVID-19). This study aimed to explore gut microbiota changes in a prospective cohort of COVID-19 children and their asymptomatic caregivers infected with the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) Omicron variant. METHODS: A total of 186 participants, including 59 COVID-19 children, 50 asymptomatic adult caregivers, 52 healthy children (HC), and 25 healthy adults (HA), were recruited between 15 April and 31 May 2022. The gut microbiota composition was determined by 16S rRNA gene sequencing in fecal samples collected from the participants. Gut microbiota functional profiling was performed by using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) software. RESULTS: The gut microbiota analysis of beta diversity revealed that the fecal microbial community of COVID-19 children remained far distantly related to HC. The relative abundances of the phyla Actinobacteria and Firmicutes were decreased, whereas Bacteroidetes, Proteobacteria, and Verrucomicrobiota were increased in COVID-19 children. Feces from COVID-19 children exhibited notably lower abundances of the genera Blautia, Bifidobacterium, Fusicatenibacter, Streptococcus, and Romboutsia and higher abundances of the genera Prevotella, Lachnoclostridium, Escherichia-Shigella, and Bacteroides than those from HC. The enterotype distributions of COVID-19 children were characterized by a high prevalence of enterotype Bacteroides. Similar changes in gut microbiota compositions were observed in asymptomatic caregivers. Furthermore, the microbial metabolic activities of KEGG (Kyoto Encyclopedia of Genes and Genomes) and COG (cluster of orthologous groups of proteins) pathways were perturbed in feces from subjects infected with the SARS-CoV-2 Omicron variant. CONCLUSION: Our data reveal altered gut microbiota compositions in both COVID-19 children and their asymptomatic caregivers infected with the SARS-CoV-2 Omicron variant, which further implicates the critical role of gut microbiota in COVID-19 pathogenesis.

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The global pandemic of coronavirus disease 2019 (COVID-19) has brought great challenges to the traditional medical model. During the outbreak of COVID-19 in Shanghai, China, from March to May, 2022, there was a significant increase in the number of pediatric cases due to high transmissibility, immune escape, and vaccine breakthrough capacity of Omicron variants. The designated hospitals for children with COVID-19 served as a connecting link between children's specialized hospitals and mobile cabin hospitals. From April 7 to June 2, 2022, a total of 871 children with COVID-19 were admitted to Renji Hospital, Shanghai Jiao Tong University School of Medicine (South Branch), a designated hospital for children with COVID-19. Among these patients, 568 (65.2%) were children under 3 years old, 870 (99.9%) were mild or moderate, and 1 was severe. This article reports the experience in the management of pediatric cases in this designated hospital, which included the following aspects: establishing an optimal case-admission process; strengthening multidisciplinary standardized diagnosis and treatment; optimizing the management, warning, and rescue system for severe COVID-19; implementing family-centered nursing care; formulating an individualized traditional Chinese medicine treatment regimen; optimizing the discharge process and strengthening bed turnover; implementing strict whole-process control to reduce the risk of nosocomial infection; constructing a structured medical record system and using information platforms to adapt to the work mode of large-volume cases; conducting scientific research and sharing the experience in diagnosis and treatment.

Expression and clinical significance of lncRNA BC041954 in ovarian cancer.

Ovarian cancer (OC) is a highly lethal disease among all gynecologic malignant tumor types. Accumulating studies have indicated that certain long non-coding RNAs (lncRNAs) serve important roles in the development and progression of OC. In a previous study by our group, lncRNA BC041954 was identified as one of the most upregulated lncRNAs in OC. In the present study, the clinical significance of lncRNA BC041954 in OC was evaluated. The expression of BC041954 was detected in OC and non-tumor tissue (NT) samples using reverse transcription-quantitative PCR. Furthermore, the association between BC041954 and clinicopathological variables was analyzed using the Chi-square test. Survival was determined using Kaplan-Meier analysis. The prognostic significance of BC041954 was evaluated using univariate and multivariate logistic regression analyses. MicroRNA (miRNA)-lncRNA and miRNA-mRNA pairs were used to construct the lncRNA-miRNA-mRNA competing endogenous RNA network with an in-house Perl script. BC041954 expression was increased in 103 OC tissues as compared with that in NT tissues. Upregulated BC041954 expression was significantly associated with the International Federation of Gynecology and Obstetrics stage and distant metastasis. Kaplan-Meier analysis demonstrated that patients with high BC041954 expression had lower overall survival (OS). In the multivariate logistic regression analysis, BC041954 was also identified as an independent poor prognostic factor for OS in patients with OC. The results suggested that overexpression of the lncRNA BC041954 is a poor prognostic indicator in patients with OC.

Erratum: MicroRNA-125b Down-Regulation Mediates Endometrial Cancer Invasion by Targeting ERBB2.

The Figure 2 and Figure 4C were incorrectly published in the article titled MicroRNA-125b down-regulation mediates endometrial cancer invasion by targeting ERBB2. Chao Shang, Yan-ming Lu, Li-rong Meng, Med Sci Monit 2012; 18(4): BR149-155. 10.12659/MSM.882617. The correct Figures are as follows.

Best practice for infection prevention in pediatric respiratory clinics during the COVID-19 epidemic.

During the COVID-19 epidemic, it is important for ensuring infection prevention and control in the pediatric respiratory clinics. Herein, we introduced the practice of infection prevention and control in pediatric respiratory clinics in China. Triage measures for patients who visit respiratory clinics, quality control for pediatric respiratory clinics and other preventive measures for related examinations and treatment have been introduced in this review article.

Indwelling Intrauterine Foley Balloon Catheter for Intraoperative and Postoperative Bleeding in Cesarean Scar Pregnancy.

STUDY OBJECTIVE: Cesarean scar pregnancy (CSP) is a rare type of ectopic pregnancy, and a significant concern in the management of this condition is the control and prevention of bleeding. We aimed to determine the efficacy and value of an indwelling, intrauterine Foley balloon catheter in controlling and preventing intraoperative and postoperative bleeding in patients with CSP. DESIGN: Retrospective case series. SETTING: University-affiliated hospital. PATIENTS: Between January 1, 2015 and May 31, 2017, 70 patients presented with CSP. INTERVENTIONS: All patients underwent uterine curettage under hysteroscopic guidance and ultrasound monitoring. Patients were then assigned to 2 groups: the study group, with an indwelling Foley balloon catheter placed in the uterine cavity during surgery and retained for 24 to 48 hours, and the control group, without catheter placement. Data were collected to compare the 2 groups in terms of intraoperative and postoperative complications, surgical time, and status of menstruation resumption. MEASUREMENTS AND MAIN RESULTS: The average daily volume of postoperative blood loss during the first 3 postoperative days in the study group was 23.1 mL compared with 31.5 mL observed in the control group (p = .041). Moreover, the study group had significantly shorter average duration of bleeding (p = .027) and fewer menstruation abnormalities than the control group. Uterine ultrasonography performed after resumption of menstruation showed that none of the enrolled patients had any intrauterine abnormalities. CONCLUSIONS: The use of an indwelling, intrauterine Foley balloon catheter has positive results in the management of CSP.

Profile and validation of dysregulated long non­coding RNAs and mRNAs in ovarian cancer.

The dysregulation of long non­coding RNAs (lncRNAs) is associated with cancer development. The present study profiled differentially expressed lncRNAs in ovarian cancer (OC) versus normal ovarian tissues (NT) and investigated their potential functions in gene expression. OC tissues from 30 patients and NT specimens from 20 non­tumor patients were collected, and 5 cases of tumor and NT were subjected to lncRNA and mRNA microarray analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed. Reverse transcription­quantitative polymerase chain reaction (RT­qPCR) was used to verify microarray data in all 30 cases. There were 2,870 differentially expressed lncRNAs (795 upregulated and 2,075 downregulated) and 2,658 differentially expressed mRNAs (1,014 upregulated and 1,644 downregulated) in OC. A total of 4 upregulated and 4 downregulated lncRNAs were validated using RT­qPCR. The data demonstrated that, with the exception of ENST00000453838 and ENST00000505048, the lncRNAs were consistent with the microarray data. Another differentially expressed lncRNA (BC041954) was assessed using independent tissue samples, and results further supported the microarray data. Moreover, GO analysis showed that the upregulated genes were involved in the 'development of the cell anatomical structure' (GO: 0048856; P=5.46x10­6), 'embryo and system development' and 'multicellular organismal development' in biological processes. By contrast, the downregulated genes were involved in 'gene expression' (GO: 0010476; P=1.81x10­6), 'nitrogen compound metabolic process', 'kidney development' and the 'cellular nitrogen compound metabolic process'. These differentially expressed lncRNAs could be classified into four classes, namely, the enhancer lncRNA nearby coding gene, HOX cluster, long­intergenic non­coding RNAs (lincRNAs) nearby coding gene and Rinn lincRNAs. Coding­non­coding gene co­expression network analysis showed the interregulation of lncRNAs and mRNAs in OC development. In conclusion, dysregulated lncRNA and mRNA expression could promote OC development. Further study may validate a number as OC markers and provide novel insights into ovarian cancer biology.

Specific siRNA inhibits XIAP expression in human endometrial carcinoma cell apoptosis.

The aim of the study was to investigate the inhibitory effects of RNA interference on XIAP gene expression of human endometrial carcinoma RL95-2 cell and the cell apoptosis. Specific small interference RNA (siRNA) of XIAP was designed and composed. Transfection of siRNA was conducted in endometrial carcinoma cell line RL95-2. The XIAP gene mRNA was assessed by real-time PCR and the change of XIAP protein was assessed with Western Blotting. The cell proliferation and apoptosis was assessed by MTT and flow cytometry methods. After transfection of siRNA specifically targeting XIAP, the relative fold of mRNA transfection in the specific transfection group was (0.04 ± 0.06) and the relative protein expression was (0.590 ± 0.178), which was significantly decreased when compared with the control group (P < 0.05); the cell growth inhibition rate in the transfection group was (47.86 ± 4.46)%, which was significantly increased when compared to the control group (P < 0.05). In vitro experiment showed that synthetic siRNA could effectively inhibit the transfection and expression of XIAP gene of human endometrial carcinoma RL95-2 cell at the mRNA level and protein level, thus significantly promote the apoptosis of endometrial carcinoma cell. The mechanisms involved in the apoptosis still require further investigation.

miR-200c modulates ovarian cancer cell metastasis potential by targeting zinc finger E-box-binding homeobox 2 (ZEB2) expression.

This study was to investigate the effect of miR-200c on regulation of ovarian cancer cell metastasis potential and explore the underlying molecular events. qRT-PCR was used to analyze the level of miR-200c expression in 48 ovarian cancer and 30 normal ovarian tissue samples. pre-miR-200c was used to manipulate miR-200c expression in ovarian cancer cells for detection of changed phenotypes of tumor cells. Bioinformatics analysis was then used to predict target genes of miR-200c and GO and pathway analyses drew the miR-200c-related gene network. Luciferase reporter assay confirmed the target of miR-200c as ZEB2. Western blot was used to detect gene expressions in ovarian cancer cells. Level of miR-200c expression was much higher in ovarian cancer than in normal ovarian tissues, and miR-200c expression was inversely associated with advanced clinical stage and lymph node metastasis of ovarian cancer (p < 0.01). The database search predicted 186 miR-200c-targeting genes, and GO analysis showed that functions of these target genes were enriched in the protein binding and other biological processes. Furthermore, miR-200c expression inhibited ovarian cancer cell ES-2 migration and invasion capacity by suppression of ZEB2 expression (p < 0.01). Overexpression of miR-200c regulated E-cadherin and vimentin expression in ovarian cancer cells. This study demonstrated high miR-200c expression in ovarian cancer tissues and ZEB2 as a targeting gene of miR-200c, which mediated the effects of miR-200c on regulation of ovarian cancer cell migration and invasion capacity and epithelial-to-mesenchymal transition. Thus, targeting of miR-200c or ZEB2 may serve as a potential therapeutic strategy for control of ovarian cancer.

Atopy in children with systemic onset juvenile idiopathic arthritis (SoJIA) is associated with a worse outcome.

CONTEXT: Atopy and systemic onset juvenile idiopathic arthritis (SoJIA) are two potential outcomes of a dysregulated immune system. Although rare, SoJIA causes 60% of the morbidity of JIA patients which exhibit a wide heterogeneity of prognosis and treatment. Co-morbidities can complicate the responses to therapy. OBJECTIVE: To study the influence of co-existing atopy on the prognosis of SoJIA. MATERIALS AND METHODS: Patients diagnosed with SoJIA between Jan 2006 and Sep 2010 were screened, enrolled in this prospective cohort study, and followed for 2 years. Management of SoJIA patients was assessed by ACR Pedi30/50/70 criteria, laboratory variables, and systemic feature score. RESULTS: At disease onset, 61 SoJIA patients (34 male and 27 female) were enrolled and were divided into SoJIA patients with atopy (n = 27) or those without atopy (n = 34). Atopic group at disease onset had significantly higher numbers of affected joints, ferritin levels and IgE serum levels than the non-atopic group. At 3 and 6 months, fewer SoJIA patients with atopy reached the ACR Pedi50 criteria (p < 0.02). During the 2 years of follow-up time, the number of infections and the number of flares were significantly higher in the SoJIA with atopy group (p < 0.01). CONCLUSION: Atopy may exert an adverse influence on SoJIA, as patients with atopy had a more active disease at diagnosis and poorer outcome. This prospective study showed that the TH1/TH2 hypothesis was too simplistic to explain the interaction between atopy and SoJIA.

Suppression of HER-2 via siRNA interference promotes apoptosis and decreases metastatic potential of SKOV­3 human ovarian carcinoma cells.

The aberrant expression of human epidermal growth factor receptor-2 (HER-2) has been detected in ovarian cancer. However, the role of HER-2 in the development of ovarian cancer has not been sufficiently elucidated. The objective of this study was to determine the role of HER-2 in the apoptosis and metastasis of SKOV-3 ovarian cancer cells. SKOV-3 cells were transfected with three double­stranded small interfering RNA (siRNA) molecules that target HER-2. Various sequences were synthesized by T7 transcription in vitro to select the most effective HER-2­silencing siRNA. SKOV-3 cells were examined for growth inhibition using the MTT proliferation assay and apoptosis was assessed using flow cytometry and TUNEL assay. The Matrigel basement memebrane matrix was used to assess invasion and chemotactic mobility, as a model of tumor cell metastasis. Western blot analysis was used to detect the expression of matrix metallopeptidase-9 (MMP-9), E-cadherin, N-cadherin and vimentin. siRNA interference in HER-2 resulted in decreased cell proliferation and invasion and increased apoptosis. Western blot analysis demonstrated a marked increase in E-cadherin and MMP-9 and a reduction in N-cadherin and vimentin protein levels in the SKOV-3 cells. The suppression of HER-2 expression resulted in apoptosis and the inhibition of metastasis of SKOV-3 cells. Therefore, the overexpression of the HER-2 gene can enhance the metastatic potential of SKOV-3 cells by increasing the protein levels of MMP-9. Epithelial-mesenchymal transition may be involved in the HER-2 siRNA-induced invasion and migration of SKOV-3 cells. Taken together, these results suggest that HER-2 functions as an oncogene and may thus be an attractive therapeutic target in SKOV-3 ovarian cancer cells.

RANTES gene polymorphisms and risk of pediatric asthma: A meta-analysis.

Numerous studies have evaluated the association between regulated upon activation, normal T cells expressed and secreted (RANTES) gene polymorphisms (-403G/A and -28C/G) and risk of pediatric asthma. However, the results have been inconsistent. A meta-analysis of the association between RANTES gene polymorphisms and pediatric asthma risk was performed in the current study. A search for published literature was conducted in the Google Scholar, PubMed and the CNKI databases (January 2000 to April 2012) and seven studies were retrieved. The associations between RANTES gene polymorphisms and pediatric asthma risk were estimated by pooled odds ratio (OR) and 95% confidence interval (CI) using a fixed- or random-effects model. Meta-analysis results revealed no significant association between the -403G/A polymorphism and risk of pediatric asthma. In the subgroup analysis by ethnicity, no association was identified between the -403G/A polymorphism and pediatric asthma risk in Caucasian and Asian populations. In the -28C/G group, the meta-analysis indicated a significant association between the -28C/G polymorphism and pediatric asthma susceptibility among the total population (recessive model: OR, 1.34; 95% CI, 1.04-1.72). However, when the subgroup analysis was performed by ethnicity, no significant associations were identified in Asians and Europeans. This result suggests that the -28C/G polymorphism may not be associated with pediatric asthma risk, while the observed increase in the risk of pediatric asthma may be due to racial differences. Additional large-scale studies are required to provide conclusive evidence on the effects of RANTES gene polymorphisms on the risk of pediatric asthma.

MicroRNA-125b down-regulation mediates endometrial cancer invasion by targeting ERBB2.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding nucleotides that regulate mRNA stability and protein expression by imperfect base pairing with the 3'-untranslated region (3'UTR) of target mRNAs. Many miRNAs have been documented to be aberrantly expressed in human cancers, but the role of miRNAs in endometrioid endometrial cancer (EEC) remains poorly understood. The objective of this study was to investigate the effect of miR-125b on EEC development and to explore its molecular mechanism in EEC carcinogenesis. MATERIAL/METHODS: Real-time quantitative PCR was applied to evaluate the expression level of miRNA-125b in EEC and normal endometrium (NE) samples. The invasion ability of miR-125b in EEC HEC1B cells was analyzed by Transwell assay after pre-miR-125b or anti-miR-125b transfection. For the invasion mechanism analysis of miR-125b on HEC1B cells, miRBase, TargetScan, miRanda and PicTar were used to predict the possible target gene of miR-125b. Luciferase activities assay, cotransfection and Western blot were used to reveal that the predicted target genes of miR-125b were direct and specific. RNA interference technology was used to confirm that the invasion inhibition of miR-125b was directly induced by ERBB2. RESULTS: Our study showed that miR-125b was down-regulated in human EEC specimens compared to that in NC specimens. Over-expression of miR-125b in HEC1B cells inhibited EEC invasion and this inhibitory effect on HEC1B cells could be restored by miR-125b knock down. Mechanism analysis revealed that ERBB2 was a direct and specific target of miR-125b. The inhibitory effect on EEC cell invasion was mediated by miR-125b inhibition of the translation of a proto-oncogene, ERBB2. CONCLUSIONS: Aberrantly expressed miR-125b contributes to HEC1B cells invasion partly through directly down-regulating ERBB2 protein expression in EEC. This miRNA signature offers a novel potential therapeutic strategy for EEC.

[Association of interferon-γ gene polymorphism and risk of cervical HPV infection].

OBJECTIVE: To investigate the association of interferon (IFN) γ gene polymorphisms and risk and prognosis of HPV cervical infection. METHODS: PCR-ASP was used for detecting IFN-γ rs2430561 polymorphism in 179 HPV positive patients and 328 HPV negative normal controls. RESULTS: The frequency of A allele of 63.7% (228/358) was significantly higher than the frequency of T allele of 36.3% (130/358) in HPV positive group (P = 0.045). The frequencies were 41.3% (74/179) in AA genotype and 14.0% (25/179) in TT genotype, women carrying AA genotype increased the risk of HPV infection compare with those with TT genotype (OR = 1.784, 95% CI: 1.031 - 3.088, P = 0.039). During follow-up, the rate of HPV positive again in AA genotype was 83.8% (62/74), while TT genotype was 20.0% (5/25). In the analysis of Kaplan-Meier, the cumulative HPV negative rates of AA, TA and TT genotype exhibited significantly different (P = 0.008). The cumulative HPV negative rate of AA genotype was the lowest (1.1% - 5.9%). CONCLUSIONS: IFN-γ rs2430561 polymorphisms confer the susceptibility to HPV infection. Women with AA genotype exhibited higher risk of infection and inclined to be continuous status and recurrence after HPV infection.

KCC1 gene advances cell invasion ability by regulating ERK signaling pathway in endometrial cancer HEC-1B cell line.

INTRODUCTION: Human potassium chloride cotransporter-1 (KCC1) gene is expressed in endometrial cancer and related to metastasis of endometrial cancer. However, whether KCC1 contributes to invasion and metastasis of endometrial cancer has not been thoroughly investigated. The purpose of this study is to research the alternation effect of insulin-like growth factor I (IGF-I) on the expression of KCC1 in endometrial cancer HEC-1B cells and to explore the mechanism of how KCC1 regulates the invasion ability of HEC-1B cells through the extracellular signal-regulated kinase (ERK) signaling pathway. METHODS: First, the inhibitive effect of RNA interference to KCC1 was detected by semiquantitative reverse transcriptase-polymerase chain reaction. Western blot was used to measure expression changes of KCC1 after exposure to IGF-I in the HEC-1B cells. The change in quantity of phosphorylated ERK1/2 (p-ERK1/2) and cell invasion ability also were measured. After RNA interference and treatment with U0126, the quantity of p-ERK1/2 and the cell invasion ability were measured again. RESULTS: After the application of IGF-I on the HEC-1B cells, the expression of KCC1 and p-ERK1/2 increased dramatically, and the cell invasion ability advanced. RNA interference could inhibit the expression of KCC1, and the quantity of p-ERK1/2 and the cell invasion ability decreased even under the effect of IGF-I. Furthermore, after treatment with U0126, the cell invasion ability no longer advanced even under the effect of IGF-I either. CONCLUSIONS: Insulin-like growth factors I can induce the upregulation of KCC1 gene, and KCC1 gene participates in the invasion ability of HEC-1B cells through the ERK signaling pathway.

[Serum galectin-9 in cervical cancer].

OBJECTIVE: To investigate the expression level of Galectin-9 (Gal-9) in the serum of cervical cancer patients and analyze the clinic significance of it. METHODS: Collected sera from 29 patients with cervical cancer, 45 patients with cervical inter-epithelial lesion (CIN) and 26 normal females. Determination of Gal-9 was performed by ELISA, and in the group of patients with cervical cancer, these results were compared with the serum vascular endothelial growth factor (VEGF) and C-reactive protein (CRP). Then selecting four patients from the invasive cervical cancer group performed Gal-9 retrospective immuno-staining analysis. RESULTS: Gal-9 concentration was 27.15 ng/L in the group of patients with cervical cancer that was significantly higher compared with the patients with CIN (21.20 ng/L) and the normal females (17.43 ng/L), and the Gal-9 concentration was significantly correlated with both VEGF and CRP in cervical cancer group. Two of these patients, who had a higher Gal-9 serum level, shown more intense staining and the other two patients, with lower serum level of Gal-9, had moderate immunostaining, suggesting that at least part of serum Gal-9 might be produced by cervical cancer tissue. CONCLUSIONS: Gal-9 might play a role in the progression and invasion of cervical cancer and warrants further study.

[Influence of human epidermal growth factor receptor-2 siRNA on chemosensitivity to cisplatin of human ovarian carcinoma cells: an in vitro experiment].

OBJECTIVE: To investigate the effects of RNA interference (RNAi) targeting human epidermal growth factor receptor 2 (HER-2) gene on the change of chemosensitivity to cisplatin of ovarian carcinoma cells. Methods Three kinds of HER-2 gene targeting siRNA, HER-2 siRNA I-III, were synthesized and the best one (HER-2 siRNA III) was screened. Human ovarian carcinoma cells of the line SKOV3 were cultured randomly divided into 3 groups: HER-2 siRNA III group, transfected with HER-2 siRNA III, non-specific siRNA group, transfected with non-specific siRNA III, and control group, without transfection. Cisplatin of the concentrations of 0, 0.05, 0.2. 0.4, 0.8, 1.0, 2.0, 10, and 20 microg/ml was added into the culture fluid for 24 h. MTT method was used to detect the proliferation rate of the SKOV3 cells. Other SKOV3 cells were divided into 3 groups: siRNA group, transfected with HER-2 siRNA III, cisplatin group, exposed to cisplatin, and HER-2 siRNA III and exposed to cisplatin. Annexin V method and flow cytometry were used to detect the apoptosis of the SKOV3 cells. The HER-2 gene expression was assessed by Western blotting. The chemosensitivity of transfected cells to cisplatin was measured by MTT. Western blotting was used to detect the protein expression of apoptosis related proteins: Bcl-2, surviving, XIAP, and Smac. RESULTS: After exposed to cisplatin, the cell survival rate decreased as the dose of cisplatin increased. The proliferation rate of the SKOV3 cells transfected with HER-2 siRNA III and exposed to 1 microg/ml cisplatin was (58 +/- 5)%, significantly lower than those of the nonspecific siRNA III transfection group [(65 +/- 6)%] and the control group [(68 +/- 3)%, both P < 0.01]. No significant difference in the cell survival rate was found between the control and nonspecific groups (P > 0.05). The apoptosis rates at different time point of the HER-2 siRNA III + cisplatin group were all higher than those of the other 2 groups (all P < 0.01). The protein expression levels of the antiapoptotic proteins Bcl-2, survivin, and XIAP were of the HER-2 siRNA III + cisplatin group were significantly lower than those of the cisplatin group, and the protein expression level of the apoptotic protein Smac of the HER-2 siRNA III + cisplatin group significantly higher than that of the cisplatin group (all P < 0.05). CONCLUSIONS: HER-2 siRNA III induces cell apoptosis significantly and increases the sensitivity of the human ovarian carcinoma cells to cisplatin. The mechanism of induction of cell apoptosis by HER-2 siRNA III + cisplatin may be related to the downregulation of Bcl-2, survivin and XIAP protein and the up-regulation of Smac protein.

[The significance of K-Cl cotransport1 expression in cervical cancer tissues].

OBJECTIVE: To investigate the expression of K-Cl cotransport 1(KCC1) in cervical cancer tissues and to investigate its role in the onset and progression of cervical cancer. METHODS: Specimens of uterus were obtained from 60 patients aged 45.89 (25 approximately 73), 40 with cervical cancer, 10 with cervical intraepithelial neoplasia (CIN), and 10 with chronic cervicitis. Semi-quantitative RT-PCR was used to detect the mRNA expression. Western blotting was used to detect the protein expression of KCC1. Immunofluorescence assay was used to detect the location of KCC1 protein. RESULTS: The mRNA expression and protein expression of KCC1 were both significantly higher in the cervical cancer tissue than those in the CIN and chronic cervicitis tissues (all P < 0.05). The levels of KCC1 in the lowly differentiated cancer tissues (at grades G(2) and G(3)) were significantly higher than those in the highly differentiated cancer tissues (at grade G(1), P < 0.05). Western blotting revealed that the protein expression level of KCC1 was significantly higher in the cervical cancer tissues than in the CIN and chronic cervicitis tissues (both P < 0.05) and the protein expression levels of KCC1 in the cancer tissues at G" 2 and G(3) grades were significantly higher than that in the cancer tissues at grade G(1) (both P < 0.05). There were no significant differences in mRNA and protein expression between the early and terminal cervical cancer tissues. There were no significant differences in the mRNA and protein expression of KCC1 between the cervical cancer cases with or without lymph node metastasis. Immunofluorescence assay showed that KCC1 was located in the cellular membrane in all patients and the KCC1 expression level there was significantly higher in the cervical cancer tissues than in the tissues of CIN and chronic cervicitis. CONCLUSION: The expression of KCC1 is up-regulated in cervical cancer and it may play an important role in the onset and progression of cervical cancer.

[Inhibition of invasive and chemotactic abilities of SKOV3 cells by human epithelial growth receptor-2 small interfering RNA].

OBJECTIVE: To investigate the effects of RNA interference (RNAi) targeting human epithelial growth receptor-2 (HER-2) gene on the invasive and chemotactic ability of SKOV3 cells. METHODS: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the positive control. Lipofectamine 2000 mediated transient transfection was conducted to transmit the siRNA into SKOV3 cells. Three pairs of specifically targeted (HER-2 siRNAI, HER-2 siRNAII, HER-2 siRNAIII) sequence were selected in the coding region of HER-2 mRNA. Transfection of HER-2 siRNA was conducted with lipofectamine 2000 in ovarian carcinoma cell line SKOV3. The HER-2 gene expression was assessed by real-time PCR and western blot assays. Changes of invasive and chemotactic capacity of SKOV3 cells were measured by polycarbonates coated with or without matrigal. RESULTS: Western blot results showed that the expression of GAPDH protein was decreased in specifically transfected cells and with the increase of siRNA dose, the expression of GAPDH protein was decreased. GAPDH protein gray value in control group, different doses (0.5, 1.0, 1.5, 2.0 microg) GAPDH siRNA interference groups were 0.6855 +/- 0.0259, 0.5698 +/- 0.0275, 0.4542 +/- 0.0296, 0.3341 +/- 0.0178 and 0.1816 +/- 0.0180, respectively. There was a significant difference in each group (F = 198.126, P < 0.01). Both HER-2 siRNAII and siRNAIII could inhibit the expression of HER-2 protein. There was a significant difference in inhibition of HER-2 expression between the siRNAIII (0.1562 +/- 0.0067), siRNAII (0.2162 +/- 0.1589) groups and the other groups (F = 69.461, P < 0.01). In comparison of the relative expression levels in each dose (0.5, 1.0, 1.5, 2.0 microg) group, the difference was significant (F = 174.53, P < 0.01). The relative expression of HER-2 mRNA in HER-2 siRNA group after the 1(st), 3(rd), 6(th) interference were 0.0506 +/- 0.0017, 0.0266 +/- 0.0011 and 0.0154 +/- 0.0020, respectively. There was a decreasing trend in expression levels at different times (P < 0.01). The invasion and chemotactic capacity of SKOV3 cells were decreased after transfection of HER-2 siRNA into the cell line (F = 53.707, P < 0.01 vs F = 11.361, P < 0.01). CONCLUSIONS: HER-2 siRNAIII can silence the mRNA expression in a time and dose dependent manner. HER-2 siRNA can inhibit cell invasion and chemotactic capacity of SKOV3 cells. The application of HER-2 siRNA extends the list of available therapeutic modalities in the treatment of human ovarian cancers.