[Related factors of negative conversion time of nucleic acid in children with COVID-19].

Objective: To explore the related factors of negative conversion time (NCT) of nucleic acid in children with COVID-19. Methods: A retrospective cohort study was conducted. A total of 225 children who were diagnosed with COVID-19 and admitted to Changxing Branch of Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from April 3rd to May 31st 2022 were enrolled in the study. The infection age, gender, viral load, basic disease, clinical symptoms and information of accompanying caregivers were retrospectively analyzed. According to age, the children were divided into<3 years of age group and 3-<18 years of age group. According to the viral nucleic acid test results, the children were divided into positive accompanying caregiver group and negative accompanying caregiver group. Comparisons between groups were performed using Mann-Whitney U test or Chi-square test. Multivariate Logistic regression analysis was used to analyze the related factors of NCT of nucleic acid in children with COVID-19. Results: Among the 225 patients (120 boys and 105 girls) of age 2.8 (1.3, 6.2) years, 119 children <3 years and 106 children 3-<18 years of age, 19 cases were diagnosed with moderate COVID-19, and the other 206 cases were diagnosed with mild COVID-19. There were 141 patients in the positive accompanying caregiver group and 84 patients in the negative accompanying caregiver group.Patients 3-<18 years of age had a shorter NCT (5 (3, 7) vs.7 (4, 9) d, Z=-4.17, P<0.001) compared with patients <3 years of age. Patients in the negative accompanying caregiver group had a shorter NCT (5 (3, 7) vs.6 (4, 9) d,Z=-2.89,P=0.004) compared with patients in the positive accompanying caregiver group. Multivariate Logistic regression analysis showed that anorexia was associated with NCT of nucleic acid (OR=3.74,95%CI 1.69-8.31, P=0.001). Conclusion: Accompanying caregiver with positive nucleic acid test may prolong NCT of nucleic acid, and decreased appetite may be associated with prolonged NCT of nucleic acid in children with COVID-19.

[Analysis of withdrawal status and influencing factors in patients receiving methadone maintenance treatment in Hubei province].

Objective: To analyze the withdrawal in patients receiving methadone maintenance treatment (MMT) and its related influencing factors in Hubei province. Methods: The patients receiving MMT in clinics in Hubei province were selected from June 2006 to December 2021. The general demographic data, drug abuse history, and MMT information were collected. The survival data of patients with MMT were analyzed by the Kaplan-Meier method, and the Cox proportional hazards model was used for multivariate analysis. Results: A total of 26 716 patients receiving MMT were included in this study, and the gender ratio between men and women was 3.34∶1(20 557∶6 159). The duration of MMT was 0.01-15.72 years, and the median duration was 2.21 (95%CI: 2.16-2.26) years. At the end of the follow-up, the withdrawal proportion was 86.75% (23 175/26 716). MMT's 0-year, 2-year, 4-year, 9-year and 14-year cumulative probabilities appeared as 67.61%, 40.24%, 30.03%, 15.49% and 6.56%, respectively. Results from the Cox proportional hazards regression model showed that the factors of the withdrawal risk were higher in patients receiving MMT, including minority nationality (HR=1.66,95%CI:1.52-1.82), having jobs (HR=1.05, 95%CI:1.01-1.08), no history of compulsory isolation or detoxification (HR=1.04, 95%CI:1.01-1.09) and the enrollment in 2016-2021 (HR=1.46,95%CI:1.35-1.58). The factors of the withdrawal risk were lower in patients receiving MMT, including 60-year-olds or above (HR=0.56,95%CI:0.42-0.75), college degree or above education level (HR=0.83, 95%CI:0.75-0.91), outpatient services of other cities (HR=0.90, 95%CI:0.87-0.93), drug use for 20 years or more (HR=0.72, 95%CI:0.66-0.80), 90 mg or more per daily dosage (HR=0.73,95%CI:0.69-0.78) and the enrollment in 2011-2015 (HR=0.93,95%CI:0.89-0.97). Conclusions: The withdrawal proportions of patients receiving MMT were high in Hubei province. The withdrawal influencing factors were complex. The daily dose was an essential factor that can be intervened under the safe MMT condition, and a higher dose should be appropriately prescribed.

[Progress on economic evaluation of pneumococcal vaccine].

Review the literature on the economic evaluation of PCV-10, PCV-13 and PPSV-23 for providing references for decision-making and research in China. A total of 17 literatures were included, and the basic information to descriptive characteristics, methodology, modeling and the results were extracted. The 14 studies adopted Markov model, 2 studies adopted decision tree model and 1 study adopted probabilistic model. The cost including vaccine price and administration costs, direct medical expenses and indirect lost. All the 17 studies use QALY as the outcome, some studies also use LYG as the outcome. 9 of 13 studies (69.2%) involving people over the age of 50 concluded that pneumococcal vaccination was cost-effective. To provide effective references for decision-makers, China should collect the relevant epidemiological parameters, vaccine effect of pneumococcal disease in the Chinese population and carry out the economic evaluation of pneumococcal vaccination.

Expression of ICAM-1 in placental tissues and the association between its gene polymorphisms and pathogenesis of preeclampsia.

OBJECTIVE: The aim of this study was to explore the expression of intercellular adhesion molecule-1 (ICAM-1) in placental tissues of patients with preeclampsia, and to elucidate the association between its polymorphisms and pathogenesis of preeclampsia. PATIENTS AND METHODS: A total of 100 preeclampsia patients (Preeclampsia group) and 100 normal puerperae (Control group) were selected as research objects. The protein expression of ICAM-1 in placental tissues was detected via Western blotting and immunohistochemical staining. The single nucleotide polymorphisms (SNPs) rs134568, rs128343, and rs201931 in the promoter region of ICAM-1 were typed via conformation difference gel electrophoresis. Chi-square test was used to detect whether the distribution frequency of ICAM-1 genotype was in agreement with Hardy-Weinberg equilibrium. The associations of ICAM-1 alleles and polymorphic sites with pathogenesis of preeclampsia were analyzed as well. Finally, the correlation between GG genotype of ICAM-1 rs134568 and clinicopathological features of preeclampsia was analyzed. RESULTS: The protein expression of ICAM-1 in placental tissues was significantly higher in Preeclampsia group than that in Control group (p<0.05). ICAM-1 SNPs rs134568, rs128343 and rs201931 all met Hardy-Weinberg equilibrium (p>0.05). According to gene correlation analysis, ICAM-1 rs134568 polymorphism and alleles were associated with the pathogenesis of preeclampsia (p<0.05). However, ICAM-1 rs128343 and rs201931 polymorphisms and alleles had no associations with the pathogenesis of preeclampsia (p>0.05). Besides, systolic blood pressure, serum creatinine level and plasma albumin level showed no statistically significant differences between people with GG genotype of ICAM-1 rs134568 in Preeclampsia group and those in Control group (p>0.05). CONCLUSIONS: ICAM-1 expression increased significantly in placental tissues of patients with preeclampsia. In addition, rs134568 in the promoter region of ICAM-1 was associated with the pathogenesis of preeclampsia.

[Development of a hyperkalemia risk assessment model for patients with chronic kidney disease].

Objective: To investigate risk factors for hyperkalemia among chronic kidney disease (CKD) patients and establish a risk assessment model for predicting hyperkalemia events. Methods: Clinical data of CKD patients (stage 3 to 5) hospitalized between May 2017 and June 2020 from 14 hospitals were retrospectively collected and divided into training dataset and validation dataset through balanced random sampling. Multivariate logistic regression analysis was used to analyze risk factors for hyperkalemia in CKD patients and the factors were scored. Receiver operating characteristic (ROC) curve was plotted and the area under the curve (AUC) was calculated. Meanwhile, the cut-off value with the best sensitivity and specificity were used to verify the accuracy of the model in validation dataset. Results: A total of 847 CKD patients were enrolled and further divided into training dataset (n=675) and validation dataset (n=172). There were 555 males and 292 females, with a mean age of (57.2±15.6) years. Multivariate logistic regression analysis showed that age, CKD stage, history of heart failure, history of serum potassium ≥5.0 mmol/L, diabetes, metabolic acidosis, and use of medications that increase serum potassium levels were risk factors for causing hyperkalemia in patients with CKD. Risk assessment model was established based on these risk factors. The AUC of the ROC curve was 0.809. Using 4 as the cut-off value, the sensitivity and specificity for predicting hyperkalemia events reached 87.1% and 57.0%, respectively. Conclusion: The model established in the current study can be used for predicting hyperkalemia events in clinical practices, which offers a new way to optimize serum potassium management in patients with CKD.

[Application of autologous cartilage in rhinoplasty for patients with traumatic nasal contraction and nasal dysfunction].

Objective: To explore the application and effect of autologous cartilage in rhinoplasty for patients with traumatic hernia and nasal dysfunction. Methods: From January 2017 to April 2019,30 patients with nasal trauma admitted to Department of Otorhinolaryngology Head and Neck Surgery, Daping Hospital, Army Medical University were treated for 6 days to 14 years. All patients were accompanied by different degrees of nasal deformity and nasal ventilatory disorders. They were classified into three categories: light, medium and heavy. All of them were open at the same time. Rhinoplasty and nasal septum deviation correction. For mild to moderate nasal deformity and nasal ventilatory disorders, ear cartilage was used to modify the tip of the nose and strengthen the nasal septal cartilage support to modify the deformity of the nasal back. For severe nasal deformity and nasal ventilatory disorders, we engraved the costal cartilage into a "Y" shaped monolithic stent or flank costal cartilage to form a 2+1 or 4+1 stent, and the costal cartilage stent and the septal cartilage tail end were sutured. Reshape the nasal septum frame support and the nasal column to avoid collapse of the nose. Results: Three patients had a septal hematoma after operation and recovered normally after cleansing. All patients were followed up for 2 months to 2 years, and no complications such as nasal septum perforation and nasal bridge collapse occurred. The postoperative cure rate was 60.0% (18/30). All patients had a grade I nasal drop after surgery, and the effective rate was 100% (30/30). The patient's preoperative measurement deviation was (6.85±2.43) mm, the postoperative measurement deviation was (2.4±1.58) mm, the preoperative nasal appearance VAS score was (1.93±1.31), and the postoperative nasal appearance VAS score was (6.60±1.16), the difference in facial appearance deviation and VAS score before and after surgery was statistically significant (P<0.05). All patients had improved nasal ventilation after operation. The preoperative nasal ventilation VAS score was (1.97±1.07), the postoperative nasal ventilation VAS score was (6.53±1.04), and the difference between preoperative and postoperative nasal ventilation VAS scores was statistically significant (P<0.05). Conclusions: Autologous cartilage is effective in rhinoplasty in patients with traumatic nasal contraction and nasal dysfunction. One stage open rhinoplasty and nasal septum deviation surgery are performed to shorten the treatment time and improve the nasal appearance and nasal ventilation function. The patients got satisfaction.

[Application of chromosomal analysis for 29 cases of fetuses with nasal bone absence or hypoplasia].

Objectives: To explore the chromosomal analysis for fetuses with nasal bone absence or hypoplasia. Methods: This was a retrospective study on 29 pregnancies nasal bone absence or hypoplasia which underwent prenatal diagnosis, including chorionic villi (CV), amniotic fluid or cordocentesis.The indication of the procedures and results were evaluated. Result: Fifteen (51.7%, 15/29) of chromosome abnormities were diagnosed with indication of fetal nasal bone absence or hypoplasia, including eight with trisomy 21, two with trisomy 13, two with X chromosome aneuploidy and three with pathogenic copy number variations (CNVs) respectively.The incidence of chromosomal abnormalities in fetus with nasal bone hypoplasia (2/5, 40%) was lower than that in fetus with nasal bone absence (13/29, 54.2%). The incidence of chromosomal abnormalities in fetus with non-syndromic nasal bone absence or hypoplasia (4/13) was lower than that in fetus with syndromic nasal bone absence or hypoplasia (11/16). Conclusion: The incidence of chromosomal abnormalities in fetus with nasal bone absence or hypoplasia was higher and prenatal diagnosis was recommended for all types of the disease.For the fetus with nasal bone absence or hypoplasia, it is recommended to give priority to CNVs for prenatal diagnosis.

Protective effects ROS up-regulation on premature ovarian failure by suppressing ROS-TERT signal pathway.

OBJECTIVE: Premature ovarian failure (POF) refers to the condition of pre-onset ovarian function failure, and is one commonly occurred disease in gynecology. Its pathogenic mechanism, however, is still unclear. Early study found decreased activity of telomerase reverse transcriptase (TERT). As an important factor to suppress TERT, oxidative stress has not been studied in POF. We, thus, investigated the role of reactive oxygen species (ROS)-TERT in POF. MATERIALS AND METHODS: Rat POF model was induced by a single intraperitoneal injection of cyclophosphamide plus 12 mg/kg busulfan. Level of follicle stimulating hormone (FSH) and inhibin B was measured by enzyme-linked immunosorbent assay (ELISA), along with hematoxylin and eosin (HE) staining to confirm successful generation of models. Western blot was applied to measure TERT expression, and N-acetyl-cysteine (NAC) or TERT small interfere RNA (siRNA) was injected to suppress ROS or TERT level, followed by HE staining to observe POF condition. RESULTS: In POF model, ovary tissues showed atrophy, less follicles, and more follicular atresia, plus mesenchymal hyperplasia. FSH and inhibin B level were significantly up-regulated and down-regulated, respectively (p<0.05). In POF rat, ROS level was elevated (p<0.05) whilst TERT level was decreased. NAC inhibited ROS level and enhanced TERT expression. In contrast, TERT siRNA further aggravated POF condition. CONCLUSIONS: ROS up-regulation inhibits TERT expression, suppresses TERT activity and facilitates POF. The ROS-TERT pathway may work as the target for treating POF.

Arachidonic acid attenuates brain damage in a rat model of ischemia/reperfusion by inhibiting inflammatory response and oxidative stress.

The aim of the present study was to study the effects of arachidonic acid (ARA) in a rat brain ischemia/reperfusion model induced by middle cerebral artery occlusion (MCAO). A total of 50 rats were randomly divided into five groups: control group, MCAO group, MCAO + ARA 0.3 g/kg group, MCAO + ARA 1 g/kg group, and MCAO + ARA 3 g/kg group. The MCAO + ARA groups received ARA by intraperitoneal injection daily for 14 consecutive days, while the rats in the control and MCAO groups were given equivalent volume of saline. We detected the Morris water maze test and pathological changes to investigate the ischemia/reperfusion injury. The protein levels of tumor necrosis factor-alpha and interleukin-6 in the hippocampus were detected by enzyme-linked immunosorbent assay kits. In addition, the activities of superoxide dismutase, glutathione peroxidase, and malondialdehyde were assayed in hippocampus homogenates to evaluate the oxidative stress after ischemia/reperfusion. The results indicated that ARA administration decreased biochemical parameters of inflammation and oxidative stress. Morris water maze test and histopathological examination further verified the protective effects of ARA on ischemia/reperfusion injury rats. These findings demonstrated that ARA could protect MCAO-induced brain injury rats by inhibition of inflammation and oxidative stress, suggesting that it may have potential as a therapy for cerebral ischemia/reperfusion injury.

Development and characterization of microsatellite markers for Solen grandis using Illumina sequencing approach.

Solen grandis is an important economic and overexploited bivalve species. In order to perform its fine-scale genetic analyses, 105 pairs of microsatellites with polymorphism were identified through Illumina Hiseq platform and bioinformatic assembly technology in this study. The estimated fragment size ranged from 100 to 268 bp and the number of alleles per locus varied between 2 and 23. Observed and expected heterozygosities varied from 0.0667 to 1.0000 and 0.0966 to 0.9492, respectively. Fourteen loci deviated significantly from Hardy-Weinberg equilibrium after Bonferroni correction. These microsatellite markers developed in this study would be helpful for future genetic studies on S. grandis and closely related species.

Blood folic acid, vitamin B12, and homocysteine levels in pregnant women with fetal growth restriction.

Deficiencies in nutrients such as folic acid and vitamin B12 may play a role in fetal growth restriction (FGR). However, whether folic acid, vitamin B12, or homocysteine is associated with FGR in Chinese populations remains unclear. This study investigated the relationship between these nutrient deficiencies and FGR in pregnant Chinese women. We selected 116 mother and infant pairs, and categorized the neonates into the FGR, appropriate for gestational age, and large for gestational age groups. Birth weight, body length, head circumference, body mass index (BMI), and Rohrer's body index of the newborns were measured. Serum folic acid, vitamin B12, and homocysteine levels were measured in mothers during the first three days of their hospital stay. Results showed that the FGR group exhibited reduced folic acid and vitamin B12 levels and elevated homocysteine levels than those in the other two groups. Folic acid and vitamin B12 levels were positively correlated with birth weight, head circumference, and BMI, whereas homocysteine level was negatively correlated with these variables. The FGR ratio in the folic acid and vitamin B12 deficiency group was higher than that in the sufficiency group (χ2 = 4.717 and 4.437, P = 0.029 and 0.035, respectively). In addition, elevated homocysteine was associated with FGR (χ2 = 5.366, P = 0.021). In conclusion, we found that folic acid and vitamin B12 deficiency was associated with elevated homocysteine levels, which may increase susceptibility to FGR.

Significant prognostic values of nuclear genes encoding mitochondrial complex I subunits in tumor patients.

In cancer biology, it remains still open question concerning the oncogenic versus oncosuppressor behavior of metabolic genes, which includes those encoding mitochondrial complex I (CI) subunits. The prognostic value of nuclear genome mRNAs expression of CI subunits is to be evaluated in the tumor patients. We used the Kaplan Meier plotter database, the cBio Cancer Genomics Portal, and the Oncomine in which gene expression data and survival information were from thousands of tumor patients to assess the relevance of nuclear genome mRNAs level of CI subunits to patients' survival, as well as their alterations in gene and expression level in tumors. We presented that the relative expression level of overwhelming majority of the nuclear genes of CI subunits with survival significance (overall survival, relapse free survival, progression free survival, distant metastasis free survival, post progression survival, and first progression), had consistent effects for patients in each type of four tumors separately, including breast cancer, ovarian cancer, lung cancer, and gastric cancer. However, in gene level, frequent cumulative or individual alteration of these genes could not significantly affect patients' survival and the overexpression of the individual gene was not ubiquitous in tumors versus normal tissues. Given that reprogrammed energy metabolism was viewed as an emerging hallmark of tumor, thus tumor patients' survival might potentially to be evaluated by certain threshold for overall expression of CI subunits. Comprehensive understanding of the nuclear genome encoded CI subunits may have guiding significance for the diagnosis and prognosis in tumor patients.

[Scalp surface skin grafts in reconstruction of external auditory meatus in congenital aural atresia].

OBJECTIVE: To investigate the clinical application of scalp skin grafts in reconstruction of external auditory meatus in congenital aural atresia. METHODS: We conducted a retrospective study on 85 patients of congenital aural atresia, all of whom were unilateral, operated from March of 2008 to December of 2010 in ENT department of the Sun Yat-Sen Memorial Hospital. The patients enrolled in the study were between 6 to 37 years old (median age 12 years), 55 male and 30 female. Scalp surface graft in ipsilateral temporal region was harvested to cover the bony external auditory meatus. RESULTS: All of these scalp split-thickness skin grafts survived without necrosis, no restenosis was found in these external auditory meatus. Neither scar nor alopecia was found in the skin-harvesting region, and hairs grew well. Granulations occurred in 27 cases in the first to sixth month posteroperatively, 20 cases recovered after local treatment. In the first year, 30 cases obtained hearing improvement more than 15 dB, 36 cases gained more than 25 dB and 19 cases gained more than 35 dB. Totally 8 patients were lost in the 4 to 5 years of follow-up, 70 cases (70/77, 90.9%) developed new external auditory meatus, 7 cases (7/77, 9.1%) suffered from stenosis in different degrees, but no atresia was found in these patients. CONCLUSION: Scalp split-thickness skin grafts has significant clinical advantage in meatoplasty of congenital aural atresia.

Anti-osteoporosis activity of red yeast rice extract on ovariectomy-induced bone loss in rats.

Osteoporosis is the most common bone disease, affecting millions of people worldwide and leading to significant morbidity and high costs. Monacolin K, an extract of red yeast rice (RYR, Hongqu), plays important roles in the management of dyslipidemia, coronary heart disease, and diabetes. Our study aimed to investigate the protective effect of monacolin K on ovariectomy-induced bone loss in rats. Fifty female Sprague-Dawley rats were randomly divided into a sham-operated and five ovariectomized (OVX) groups: OVX with vehicle, OVX with fluvastatin, and OVX with RYR extract of three graded doses. Bone mineral density (BMD), biochemical markers, and cell viability were analyzed by dual energy X-ray absorptiometry, enzyme-linked immunosorbent assay, and 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Gene expression was evaluated by real-time polymerase chain reaction amplification and western blot. Our results showed that administration of RYR extract markedly increased the bone mineral density in OVX rats. Moreover, RYR extract decreased the levels of bone turnover markers, including osteocalcin and tartrate resistant acid phosphatase activity. The MMT assay revealed that RYR extract treatment significantly improved the osteoblast viabilities in a dose-dependent manner (P < 0.05). At the molecular level, we further demonstrated that RYR extract enhanced the expression of Bmp2 and Bmp4 both at the mRNA and protein levels. Collectively, these data suggested RYR extract could protect against osteoporosis in ovariectomized rats, most likely through activation of BMP2/4 expression.

Observed impacts of insulin therapy on callus cell transforming growth factor-beta 1 expression in diabetic rats.

The expression of transforming growth factor-beta 1 (TGF-ß1) inside the callus cells of diabetic rats and the impact of insulin therapy on its expression and biomechanics was investigated. The rats were randomly divided as follows: an insulin therapy group (IT), a diabetic model group (DM), and a non-diabetic control group (NC). Bone specimens from each group were extracted at different times for immunohistochemical observation of the expression of TGF-ß1. Concurrently, the destruction torque and torsional stiffness were detected at different times. One to four weeks after fracture, TGF-ß1 was widely expressed in fractured callus cells and periosteal proliferating cells, while the expression inside diabetic cells was significantly reduced. The expression of TGF-ß1 decreased over the first 68 weeks, and the mature bone cells never expressed TGF-ß1. The destruction torque (Nm) detected in the 6th week revealed that there was a statistically significant difference between the DM, NC, and IT groups (P < 0.01). In conclusion, TGF-ß1 expression was significantly reduced inside the callus cells of diabetic rats. Insulin therapy increased TGF-ß1 expression inside the callus cells of diabetic rats and improved the biomechanical characteristics of the callus.

Whole exome sequencing implicates PTCH1 and COL17A1 genes in ossification of the posterior longitudinal ligament of the cervical spine in Chinese patients.

Ossification of the posterior longitudinal ligament (OPLL) of the cervical spine is a complex multifactorial disease. Patients with OPLL commonly present with symptoms in their 40s or 50s. The genetic basis of OPLL remains poorly understood. Exome capture combined with massively parallel DNA sequencing has been proposed as an efficient strategy to search for disease-causing genes of both monogenic and multigenic disorders. To identify candidate pathogenic genes associated with OPLL, we performed whole exome sequencing (WES) on two unrelated southern Chinese OPLL patients. The entire DNA coding region of the candidate genes was amplified by PCR and Sanger sequenced. The common single nucleotide polymorphisms were analyzed by association studies. WES revealed p.T265S/PTCH1, p.P1232L/PTCH1, and p.T902S/COL17A1 mutants in the two female cases with mixed OPLL. These were confirmed by Sanger sequencing. p.P1232L/PTCH1, p.N1374D/COL17A1 and p.T902S/COL17A1 were subsequently identified in three males with continuous OPLL and one female with mixed OPLL. The association studies indicated that the SNPs rs805698 and rs4918079 in COL17A1 were significantly associated with OPLL. This study suggests that WES may be a practical approach to revealing significant genetic involvement in OPLL. Variants of the PTCH1 and COL17A1 genes may contribute to the development of OPLL.

Prophylactic thyroidectomy for MEN 2-related medullary thyroid carcinoma based on predictive testing for RET proto-oncogene mutation and basal serum calcitonin in China.

INTRODUCTION: Early and normative surgery is the only curative method for multiple endocrine neoplasia type 2 (MEN 2)-related medullary thyroid carcinoma (MTC). AIMS: To study the timing of prophylactic total thyroidectomy (TT) for MEN 2-related MTC with different RET mutations in a Chinese population, and to compare the sensitivity and accuracy of fully-automated chemiluminescence immunoassay (FACLIA) and radioimmunoassay (RIA) for serum calcitonin (Ct). METHODS: We collected 24 asymptomatic individuals from 8 unrelated Chinese families with MEN 2, and analyzed RET mutation and Ct levels. Then we performed TT on 17 of the 24 individuals, including TT (2/17), TT with bilateral level VI lymph-node dissection (B-LND(VI); 12/17) and TT with B-LND(VI) + modified unilateral/bilateral/local neck dissection (3/17). RESULTS: Histopathology revealed bilateral/unilateral MTC in 15/17 (88.2%; median diameter, 1.0 cm) and bilateral C-cell hyperplasia in 2/17 (11.8%; p.V292M/R67H/R982C and p.C618Y). Lymph-node metastasis/fibro-adipose tissue invasion (p.C634R) or solely fibro-adipose tissue invasion (p.C634Y) were found in 2/17 (11.8%). Elevated pre-surgical Ct (pre-Ct) was identified by FACLIA in 17/17 (median age, 24.0), while pre-Ct by RIA was found in only 6/15 (P < 0.001). The median follow-up was 22.0 months, during which 16/17 had no abnormality (one p.C634R individual had elevated Ct), and another 7 carriers still had consistently undetectable Ct by FACLIA. CONCLUSIONS: Our study highlights the importance and feasibility of individualized prophylactic TT for MEN 2-related MTC, based on predictive integrated screening of RET and pre-Ct levels. Besides, we recommend FACLIA to measure Ct for earlier diagnosis, treatment and follow-up monitoring of MTC.

Co-delivery of LETM1 and CTMP synergistically inhibits tumor growth in H-ras12V liver cancer model mice.

As hepatocellular carcinoma (HCC) is one of the most common tumors worldwide, development of novel therapeutic approaches for HCC is urgently needed. Two different genes, LETM1 and CTMP, which target mitochondrial functions, were chosen and linked using 2A-peptide sequence. Successful self-cleavage of 2A-peptide induced synergistic antitumor effect in the liver of H-ras12V, the HCC model mice, by simultaneous activation of LETM1 (Leucine zipper/EF hand-containing transmembrane-1) and CTMP (carboxyl-terminal modulator protein). Overexpression of LETM1 and CTMP significantly reduced the incidence of tumorigenesis, which were confirmed by gross and microscopic observations. Morphological changes in mitochondria, such as swelling and loss of cristae, were significant, and the prolonged activation of defects in mitochondrial function led to mitochondria-mediated apoptosis. Furthermore, with CTMP as a direct binding partner of Akt1, and LETM1 as a binding partner of CTMP, LETM1-2A-CTMP downregulated the Akt1 pathway at both Ser473 and Thr308 sites of phosphorylation. Proliferation and angiogenesis, which are important in cancer prognosis, were reduced in tumor sites after introduction of LETM1-2A-CTMP. Taken together, the results indicate that introduction of the mitochondria-targeting genes, LETM1 and CTMP, and self-processing capacity of 2A-peptide sequence exerts an antitumor effect in liver of H-ras12V mice, suggesting its potential as a tool for gene therapy.

Suppression of tumor growth in xenograft model mice by programmed cell death 4 gene delivery using folate-PEG-baculovirus.

Cancer gene therapy using tumor suppressor genes is considered to be an attractive approach for arresting cell growth and inducing apoptosis. Programmed cell death 4 (Pdcd4) is a tumor suppressor gene, which prevents tumorigenesis and tumor progression. To address the issue of whether expression of PDCD4 protein induces apoptosis in cancerous cells, the Pdcd4 gene was delivered using folate-PEG-baculovirus. Folate-PEG-baculovirus containing Pdcd4 gene (F-P-Bac-Pdcd4) was constructed by attachment of F-PEG to the baculovirus surface using chemical modification. The F-P-Bac-Pdcd4 showed enhanced transduction efficiency, efficiently expressed PDCD4 protein, and induced apoptosis in human epidermal carcinoma (KB) cells as compared with an unmodified baculovirus. In a tumor xenograft study, injection of F-P-Bac-Pdcd4 into tumors established from the KB cell line by subcutaneous implantation significantly suppressed tumor growth and induced apoptosis. Thus, this study shows a new baculovirus-mediated tumor suppressor gene delivery system for cancer therapy.

Degradable polyethylenimines as DNA and small interfering RNA carriers.

Gene therapy is a powerful approach in the treatment of a wide range of both inherited and acquired diseases. Nonviral delivery systems have been proposed as safer alternatives to viral vectors because they avoid the inherent immunogenicity and production problems that are seen when viral systems are used. Many cationic polymers, including high-molecular-weight polyethylenimine (PEI) have been widely studied as gene-delivery carriers, both, in vitro and in vivo. However, interest has recently developed in degradable polymeric systems. The advantage of degradable polymer is its low in-vivo cytotoxicity, which is a result of its easy elimination from the cells and body. Degradable polymer also enhances transfection of DNA or small interfering RNA (siRNA) for efficient gene expression or silencing, respectively. This review paper summarizes and discusses the recent advances with degradable PEIs, such as cross-linked and grafted PEIs for DNA and siRNA delivery.