The expression of UNC5D is abnormal in the early stage of colorectal tumors associated with its proliferation and migration.

OBJECTIVE: Colorectal adenomas are an important precancerous lesion of colorectal adenoma with a high incidence. This study aims to explore new prognostic targets for colorectal adenomas through bioinformatics techniques. MATERIALS AND METHODS: In this study, data from 29 colonic adenomas and 38 normal colonic mucosa in GSE37364 were analyzed to screen for differentially expressed genes (DEGs). Then, batch survival analysis, construction of risk model, mutation analysis, Cox regression analysis and expression analysis were performed on DEGs to determine the hub genes of this study. Finally, immune correlation analysis and cell experiments were carried out on the hub gene to explore its potential mechanism. RESULTS: In our study, a total of 431 up-regulated and 809 down-regulated differentially expressed genes (DEGs) were identified. Among these, Unc-5 Netrin Receptor D (UNC5D) emerged as a pivotal gene associated with colorectal adenoma. Notably, UNC5D expression levels were found to be significantly higher in normal tissues compared to colorectal adenoma tissues. Furthermore, our analysis demonstrated that UNC5D showed promising diagnostic potential for patients with colon adenocarcinoma. In vitro experiments revealed that the overexpression of UNC5D had a profound impact on the behavior of colorectal tumor cells. Specifically, it led to a substantial reduction in the proliferation, motility, and invasion of these tumor cells. Additionally, UNC5D was shown to exert control over STAT1/STAT3 phosphorylation, which in turn regulated the expression of PD-L1 in response to interferon (IFN) stimulation. These findings highlight the significant role of UNC5D in modulating immune responses and the development of colorectal adenoma. UNC5D emerges as a potential diagnostic biomarker and an attractive immunotherapeutic target in the context of colorectal malignancies. These results call for further exploration of UNC5D-based strategies for the diagnosis and treatment of colorectal adenoma and adenocarcinoma. CONCLUSIONS: In addition to having the potential to be used as a diagnostic biomarker and an immunotherapeutic target in colorectal malignancies, UNC5D is necessary for the growth of colorectal adenomas. Additionally, UNC5D controlled STAT1/STAT3 phosphorylation to suppress the growth of colorectal cancers by regulating IFN-induced PD-L1 expression.

[Comparative study of next generation sequencing and immunohistochemistry on molecular classification of endometrial carcinoma].

Objective: To investigate the differences in molecular classification of endometrial carcinoma (EC) between various technical methods and to explore molecular classification schemes suitable for Chinese population. Methods: The study used a comprehensive scheme of next generation sequencing (NGS) and immunohistochemistry for molecular classification of 254 EC cases that were obtained at Department of Pathology, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, China from April 2021 to March 2022. According to the recommended threshold of Sanger sequencing which was approximate-20% variant allele fraction (VAF), NGS data were extracted to simulate the results of Sanger sequencing. Results: The 254 EC patients had a mean age of 51 years (range, 24 to 89 years). Combination of POLE (9-14 exons), TP53 total exons and microsatellite instability (MSI) detection was a better single scheme than NGS alone, while combination of MSI fragment analysis and conventional immunohistochemistry was the best solution and seemed best aligned with TCGA data and recent studies. POLE ultramuted type, mismatch repair defect type, TP53 mutant type and non-specific molecular characteristic type accounted for 11.4% (29/254), 31.5% (80/254), 22.4% (57/254) and 34.6% (88/254) of the cases, respectively. If Sanger sequencing was adopted for POLE and TP53 detection, the frequencies of these EC types were 9.1% (23/254), 31.5% (80/254), 12.9% (33/254) and 46.6% (118/254), respectively, with greatly increasing non-specific molecular characteristics cases. If POLE was detected by Sanger sequencing and others by immunohistochemistry, they were 9.1% (23/254), 42.2% (92/218), 13.8% (35/254) and 40.9% (105/254), respectively, with increasing the false positive rates of the mismatch repair defect group. Conclusions: Small and medium-sized NGS panels with MSI detection is a better solution than NGS alone. Sanger sequencing is currently available for POLE mutation detection, which is not sensitive enough for TP53 mutation detection, and seems equivalent to the efficiency of TP53 by immunohistochemistry. Further optimization of small and medium-sized NGS panels covering MSI detection and POLE and TP53 full exons may be the best choice for the future to meet national conditions.

[Value of high-risk HPV E6/E7 mRNA in situ hybridization in grading of cervical intraepithelial neoplasia].

Objective: To investigate the value of high-risk HPV E6/E7 mRNA in situ hybridization in the grading of cervical squamous intraepithelial neoplasia (CIN). Methods: A total of 261 cases with diagnosis of CIN and cervical squamous cell carcinoma (SCC) at west China University Second Hospital, Sichuan University from July 2019 to June 2020 were collected, including 60 cases of CIN1, 41 cases of CIN2, 51 cases of CIN3, 72 cases of SCC, and 37 cases of normal cervical control tissue (10 HPV negative and 27 HPV positive). All pathologic tissues were made into tissue microarrays, and HE staining, HPV E6/E7 mRNA in situ hybridization (ISH) and p16 immunohistochemical (IHC) staining were performed respectively. The staining was assessed by light microscopy, and the positive rate and positive pattern were analyzed statistically. Results: HPV mRNA ISH in CIN1 mainly showed spot staining in predominantly basal to mid-epithelial layers (≤BME) with a diffuse nuclear signals in the superficial layer (supD), that is, the pattern of ≤BME+supD; in CIN2, it mainly showed spot staining in predominantly basal to above mid-epithelial but not the full layer (>BME) and some cases with supD staining, that is, the pattern of>BME+supD; In CIN3, the mainly pattern was >BME, and the spot staining was distributed throughout the epithelium. In CIN1, CIN2 and CIN3, there were significantly statistical differences among the above three staining patterns (P<0.05). Conclusions: HPV mRNA ISH contributes to the accurate diagnosis and grading of CIN, and has better specificity than p16 IHC staining.

[Clinical follow-up analysis of nusinersen in the disease-modifying treatment of pediatric spinal muscular atrophy].

Objective: To explore the clinical efficacy of disease-modifying drug nusinersen on children with spinal muscular atrophy. Methods: The baseline and longitudinal clinical data of 15 children who were treated with nusinersen in the Children's Hospital, Zhejiang University School of Medicine from October 2019 to October 2021 were retrospectively collected. The general data (gender, age, genotype, and clinical classification, etc.), motor function, nutritional status, scoliosis and respiratory function were analyzed. Wilcoxon rank-sum test was used for comparing multi-system conditions before and after treatment. Results: The age of 15 cases (7 males, 8 females) was 6.8 (2.8, 8.3) years, with 2 cases of type 1, 6 cases of type 2, and 7 cases of type 3 respectively, and the course of disease was 55.0 (21.0, 69.0) months. After 9.0 (9.0, 24.0) months of treatment, the motor function scale evaluations of the Hammersmith neurological examination section 2 (13.0 (7.0, 23.0) vs. 18.0 (10.0, 25.0) scores, Z=-2.67, P=0.018) of 15 children, the Hammersmith functional motor scale expanded (38.0 (18.5, 45.5) vs. 42.0 (23.0, 51.0) scores, Z=-2.38, P=0.018), and the revised upper limb module (27.0 (19.5, 32.0) vs. 33.0 (22.5, 35.5) scores, Z=-2.52, P=0.012) of children with type 2 and 3 had significantly improved. Thirteen patients achieved clinically significant motor function improvement, and 2 of them had kept stable scale scores. Subjective reports also indicated that the muscle strength and daily exercise ability of these children improved after treatment, and no serious adverse reactions were reported. Supplemented by the multi-disciplinary team management, the levels of some indicators such as Cobbs angle of scoliosis and forced vital capacity all had significantly improved (all P<0.05). Conclusions: Nusinersen can improve the motor function of patients with 5q spinal muscular atrophy, which is also proved safe to be used in children. The drug treatment supplemented by the multi-disciplinary team management is helpful to improve the multi-system function of the children with spinal muscular atrophy.

[Idiopathic pleuroparenehymal fibroelastosis: five case reports and review of literature].

Objective: To raise the awareness of idiopathic pleuroparenehymal fibroelastosis (iPPFE) through investigating the clinical, radiographic and pathological features. Methods: Five cases of iPPFE proved by pathology. The clinical data were studied respectively, and the relevant literature was reviewed. Results: All the cases of iPPFE were manifested by cough and dyspnea. The patients including 3 males and 2 females, aged from 30 to 70 years Chest CT scan showed pleural thickening, subpleural consolidation in both upper lungs complicated with tractive bronchiectasis.Computed tomography-guided percutaneous lung biopsy or surgical lung were performed and the same pathological showed pleura and subpleural dense elastic and collagen fibers. The elastic fibers stain was also positive,which was consistent with PPFE. One patient received low-dose corticosteroid, two received pirfenidone therapy, the others received no treatment. Three patients were stable during the follow-up. Conclusions: iPPFE has characteristic pathological features. However, the number of clinically reported cases is low due to missed diagnosis or misdiagnosed. Improving the understanding of features of iPPFE is helpful for the dianosis, therapy, and prognosis of this disease.

Activation of the NLRP3 and AIM2 inflammasomes in a mouse model of Schistosoma mansoni infection.

Schistosomiasis is an inflammatory disease that occurs when schistosome species eggs are deposited in the liver, resulting in fibrosis and portal hypertension. Schistosomes can interact with host inflammasomes to elicit host immune responses, leading to mitochondrial damage, generation of high levels of reactive oxygen species (ROS) and activation of apoptosis during inflammation. This study aims to examine whether ROS and NF-κB (p65) expression elicited other types of inflammasome activation in Schistosoma mansoni-infected mouse livers. We examine the relationship between inflammasome activation, mitochondrial damage and ROS production in mouse livers infected with S. mansoni. We demonstrate a significant release of ROS and superoxides and increased NF-κB (p65) in S. mansoni-infected mouse livers. Moreover, activation of the NLRP3 and AIM2 inflammasomes was triggered by S. mansoni infection. Stimulation of HuH-7 hepatocellular carcinoma cells with soluble egg antigen induced activation of the AIM2 inflammasome pathway. In this study, we demonstrate that S. mansoni infection promotes both NLRP3 and AIM2 inflammasome activation.

Ligustilide, a major bioactive component of Angelica sinensis, promotes bone formation via the GPR30/EGFR pathway.

Angelica sinensis (Oliv.) Diels is a widely-used traditional Chinese herbal medicine in treating osteoporosis. Ligustilide (LIG) is the main component of A. sinensis and is considered to be the most effective biologically active ingredient in this plant. LIG has been found to have multiple pharmacological activities, such as anti-atherosclerosis, neuroprotection, anticancer, anti-inflammatory and analgesic. However, little is known regarding its anti-osteoporotic effects. The aims of this study were to investigate any protective effect of LIG on bone formation. The results showed that LIG significantly ameliorated inhibition of bone formation in zebrafish caused by prednisolone. LIG promoted osteoblast differentiation, including that of the pre-osteoblastic cell line MC3T3-E1 and bone marrow mesenchymal stem cells. LIG greatly improved the viability of MC3T3-E1 cells exposed to H2O2, attenuated H2O2-induced apoptosis and increased the expression of Bcl-2. Furthermore, LIG treatment lead to marked activation of phosphorylated EGFR and ERK1/2. These effects could be obviously inhibited by blocking GPR30 signaling with the specific inhibitor G15. Collectively, the results reveal that GPR30 is a positive switch for LIG to increase bone formation via regulation of EGFR, and these results provide evidence for the potential of LIG to treat osteoporosis.

[Association between C1114G polymorphism in the regulator of G protein signaling 2 and vasovagal syncope in children].

Objective: To analyze the distribution of the regulator of G protein signaling 2 (RGS2) gene C1114G polymorphism in children with vasovagal syncope (VVS) and the associated clinical classification groups, and to explore the association between RGS2 C1114G and VVS. Methods: This was a prospective case-control study. A head-up tilt test (HUT) was performed in 300 children visiting Children's Hospital Affiliated to Shanghai Jiaotong University from August 2010 to December 2015 for unexplained syncope. A total of 150 children with positive HUT and a diagnosis of VVS were enrolled and assigned to the VVS group. The VVS group was further divided into 3 subgroups based on characteristics of the heart rate and blood pressure measured during the HUT. A total of 150 children with negative HUT were enrolled and assigned to the HUT-negative group. A total of 150 healthy children were enrolled as the normal control group for genetic polymorphism detection. The clinical characteristics of patients in the VVS group and the HUT-negative group were recorded. Peripheral blood samples of each case were collected. RGS2 C1114G polymorphism was evaluated using high-resolution melting curve and polymerase chain reaction together with gene sequencing. The genotype and allele frequency were analyzed and compared among different groups (VVS, HUT-negative, and normal control) and VVS subgroups. Comparisons among groups were performed using Chi-square test. Results: Patients in the VVS group (48 males and 102 females, aged (10.1±3.2) years) were more frequently female (68.0% vs. 57.3%;χ(2)=5.090, P=0.024) compared with patients in the HUT-negative group (67 males and 83 females, aged (10.8±2.2) years). No significant difference was found regarding the distribution of the CC genotype, CG genotype and GG genotype among the VVS group (n=98, 65.3%; n=36, 24.0%; n=16, 10.7%), the HUT-negative group (n=112, 74.7%; n=28, 18.7%; n=10, 6.7%) and the normal control group (n=108, 72.0%; n=31, 20.7%; n=11, 7.3%) (χ(2)=3.632, P=0.458). There was no significant difference in the frequencies of C allele and G allele in the VVS group (n=232, 77.3%; n=68, 22.7%), the HUT-negative group (n=252, 84.0%; n=48,16.0%) and the normal control group (n=247, 82.3%; n=53, 17.7%) (χ(2)=4.659, P=0.097). The 150 children in the VVS group were further divided into the mixed-response subgroup (n=83), vasodepressor-response subgroup (n=42) and cardioinhibitory-response subgroup (n=25). The CC genotype, CG genotype and GG genotype in the mixed-response subgroup, the vasodepressor-response subgroup and the cardioinhibitory-response subgroup were (n=65, 78.3%; n=16, 19.3%; n=2, 2.4%), (n=20, 47.6%; n=11, 26.2%; n=11, 26.2%) and (n=13, 52.0%; n=9, 36.0%; n=3, 12.0%), respectively. The frequencies of C allele and G allele in the mixed-response subgroup, the vasodepressor-response subgroup, and the cardioinhibitory-response subgroup were (n=146, 88.0%; n=20, 12.0%), (n=51, 60.7%; n=33, 39.3%) and (n=35, 70.0%; n=15, 30.0%), respectively. The percentages of the GG genotype and G allele were significantly higher in the vasodepressor-response subgroup than the other two subgroups (χ(2)=21.698, 25.345, all P=0.000). Conclusions: No significant association was found between RGS2 C1114G polymorphism and VVS in children. Due to the higher distribution of GG genotype and G allele in the vasopressor-response subgroup, RGS2 C1114G may be associated with the regulation of blood pressure during the onset of VVS in children.

[Comparison between computer aided simulation and dental model orthognathic surgery for the treatment of patients with mandibular excess and facial asymmetries].

Objective: To evaluate the results of the orthognathic surgery with computer aided simulation-three-dimensional(3D) printed surgical guide and dental model surgery in the treatmemt of patients with mandibular excess and facial asymmetries. Methods: The coordinate system was built in ProPlan CMF 2.0 software, and the horizontal plane consisted of PoL, PoR, OrL, middle sagittal plane through nasion point and basion point and the plane perpendicular to the horizontal plane, coronoid plane through nasion point and the plane perpendicular to the horizontal plane and middle sagittal plane. The orientation of maxilla and mandibular distal segment was calculated by each triangle(U1-U6L-U6R, L1-L6L-L6R, Me-M5L-M5R) before and after orthognathic surgery. A total of 60 mandibular excess patients with facial asymmetries were enrolled in this retrospective study. They were divided into two groups, group Ⅰ with computer aided simulation, group Ⅱ with dental model surgery. The difference of maxillary occlusal plane roll and yaw angle, mandibular occlusal plane roll and yaw angle, and mandibular body plane roll and yaw angle were calculated. Statistical analysis was performed with SPSS 17.0 software. Results: The yaw angle of mandibular occlusal plane of the dental model surgery and computer aided simulation was 0.36°± 0.48° and 0.84° ± 0.36° (P=0.043), respectively. The roll angle of mandibular occlusal plane of the dental model surgery and computer aided simulation was 0.26°±0.79° and 0.54°±0.40°(P=0.032), respectively. The yaw angle of mandibular body plane of the dental model surgery and computer aided simulation was 0.60°± 1.04° and 0.23°±0.52°(P=0.008), respectively. The roll angle of mandibular body plane of the dental model surgery and computer aided simulation was 0.82° ± 0.72° and 0.53° ± 0.37° (P=0.028), respectively. The changes in computer aided simulation group were more obvious than that in the dental model surgery group, but the difference was not significant in the yaw angle of maxillary occlusal plane and the roll angle of maxillary occlusal plane between the two groups(P >0.05). Conclusions: It was more effective to correct mandibular asymmetry by computer aided simulation than dental model surgery.

Minimizing radiation exposure during percutaneous nephrolithotomy.

Given the recent trends in growing per capita radiation dose from medical sources, there have been increasing concerns over patient radiation exposure. Patients with kidney stones undergoing percutaneous nephrolithotomy (PNL) are at particular risk for high radiation exposure. There exist several risk factors for increased radiation exposure during PNL which include high Body Mass Index, multiple access tracts, and increased stone burden. We herein review recent trends in radiation exposure, radiation exposure during PNL to both patients and urologists, and various approaches to reduce radiation exposure. We discuss incorporating the principles of As Low As reasonably Achievable (ALARA) into clinical practice and review imaging techniques such as ultrasound and air contrast to guide PNL access. Alternative surgical techniques and approaches to reducing radiation exposure, including retrograde intra-renal surgery, retrograde nephrostomy, endoscopic-guided PNL, and minimally invasive PNL, are also highlighted. It is important for urologists to be aware of these concepts and techniques when treating stone patients with PNL. The discussions outlined will assist urologists in providing patient counseling and high quality of care.

TRAF2 is a biologically important necroptosis suppressor.

Tumor necrosis factor α (TNFα) triggers necroptotic cell death through an intracellular signaling complex containing receptor-interacting protein kinase (RIPK) 1 and RIPK3, called the necrosome. RIPK1 phosphorylates RIPK3, which phosphorylates the pseudokinase mixed lineage kinase-domain-like (MLKL)-driving its oligomerization and membrane-disrupting necroptotic activity. Here, we show that TNF receptor-associated factor 2 (TRAF2)-previously implicated in apoptosis suppression-also inhibits necroptotic signaling by TNFα. TRAF2 disruption in mouse fibroblasts augmented TNFα-driven necrosome formation and RIPK3-MLKL association, promoting necroptosis. TRAF2 constitutively associated with MLKL, whereas TNFα reversed this via cylindromatosis-dependent TRAF2 deubiquitination. Ectopic interaction of TRAF2 and MLKL required the C-terminal portion but not the N-terminal, RING, or CIM region of TRAF2. Induced TRAF2 knockout (KO) in adult mice caused rapid lethality, in conjunction with increased hepatic necrosome assembly. By contrast, TRAF2 KO on a RIPK3 KO background caused delayed mortality, in concert with elevated intestinal caspase-8 protein and activity. Combined injection of TNFR1-Fc, Fas-Fc and DR5-Fc decoys prevented death upon TRAF2 KO. However, Fas-Fc and DR5-Fc were ineffective, whereas TNFR1-Fc and interferon α receptor (IFNAR1)-Fc were partially protective against lethality upon combined TRAF2 and RIPK3 KO. These results identify TRAF2 as an important biological suppressor of necroptosis in vitro and in vivo.

Prevalence and genotype of Giardia duodenalis from faecal samples of stray dogs in Hualien city of eastern Taiwan.

Giardia duodenalis is a zoonotic protozoan parasite that causes diarrhea through waterborne transmission or fecal-oral infection. The cysts are chlorine-resistant and, therefore, can pollute drinking water and induce a pandemic disease. In this study, we aimed to detect G. duodenalis infection in stray dogs in Hualien, Taiwan. We collected faecal samples from 118 dogs and amplified DNA sequences of the ß-giardin gene by nested polymerase chain reactions (nested PCR). Eleven of the 118 faecal samples tested positive for the parasite. The genotype analysis of the 11 samples indicated that 7 samples belonged to assemblage C and four samples belonged to assemblage D. Our study provided a better understanding of the infection rate and genotypes of G. duodenalis in dogs from Hualien City, and human infection could not be induced by this zoonotic infection pathway in Hualien City.

Efficacy and safety of ipilimumab in metastatic melanoma patients surviving more than 2 years following treatment in a phase III trial (MDX010-20).

BACKGROUND: In a phase III trial (ClinicalTrials.gov registration ID: NCT00094653), ipilimumab significantly improved survival versus a vaccine control in pretreated patients with metastatic melanoma. Here, we characterize outcomes of those patients who survived ≥ 2 years. METHODS: Patients were randomized (3 : 1 : 1) to receive ipilimumab 3 mg/kg + gp100 vaccine, ipilimumab 3 mg/kg + placebo, or gp100 vaccine alone. Baseline demographic data, duration of survival, responses, and safety among patients with ≥ 2 years' survival were analyzed. RESULTS: Among 676 randomized patients, 474 and 259 patients had at least 2 or 3 years of potential follow-up, respectively, and were eligible for analysis. Among these, 94 (20%) and 42 (16%) survived ≥ 2 and ≥ 3 years, respectively. Survival rates at 2 and 3 years were 25% (24 of 95) and 25% (13 of 53) with ipilimumab alone and 19% (54 of 284) and 15% (24 of 156) with ipilimumab plus gp100. Safety among patients with ≥ 2 years' survival was comparable with the overall study population, with the onset of new ipilimumab-related toxic effect (all grades) reported in 6 of 78 (8%) patients. CONCLUSIONS: Ipilimumab results in survival of ≥ 2 years in one-fifth of pretreated patients with 2 years potential follow-up in a phase III trial. New onset, low-grade events starting after administration of the last dose were infrequent. TRIAL REGISTRATION ID: NCT00094653.

Missing data and censoring in the analysis of progression-free survival in oncology clinical trials.

Progression-free survival (PFS) is increasingly used as a primary endpoint in oncology clinical trials. However, trial conduct is often such that PFS data on some patients may be partially missing either due to incomplete follow-up for progression, or due to data that may be collected but confounded by patients stopping randomized therapy or starting alternative therapy prior to progression. Regulatory guidance on how to handle these patients in the analysis and whether to censor these patients differs between agencies. We present results of a reanalysis of 28 Phase III trials from 12 companies or institutions performed by the Pharmaceutical Research and Manufacturers Association-sponsored PFS Expert Team. We show that analyses not adhering to the intention-to-treat principle tend to give hazard ratio estimates further from unity and describe several factors associated with this shift. We present illustrative simulations to support these findings and provide recommendations for the analysis of PFS.

The coding-synonymous polymorphism rs1045280 (Ser280Ser) in beta-arrestin 2 (ARRB2) gene is associated with tardive dyskinesia in Chinese patients with schizophrenia.

BACKGROUND: Tardive dyskinesia (TD) is a severe and potentially irreversible adverse effect of long-term antipsychotic treatment. Typical antipsychotics are commonly binding to the dopamine receptor D2 (DRD2), but the occurrence of antipsychotic-induced TD is rather delayed; therefore, the development of TD may be associated with mediators or signalling complexes behind DRD2, such as beta-arrestin 2 (ARRB2), an important mediator between DRD2 and serine-threonine protein kinase (AKT) signal cascade. METHODS: A case-control study to evaluate the association between rs1045280 (Ser280Ser) and antipsychotic-induced TD was performed amongst 381 patients (TD/non-TD = 228/153). RESULTS: There was a significant difference in the genotype distribution between TD and non-TD groups (P = 0.025); furthermore, the allelic analysis indicated that patients with T allele had increased risk of TD occurrence (OR(T) = 1.58, 95% CI = 1.14-2.19, P = 0.007). CONCLUSIONS: To the best of our knowledge, this is the first study reporting a positive association between the SNP rs1045280 and TD in schizophrenic patients.

Tetracycline release from tripolyphosphate-chitosan cross-linked sponge: a preliminary in vitro study.

BACKGROUND AND OBJECTIVE: The aim of this study was to design a tripolyphosphate-chitosan cross-linked tetracycline-containing (TPP-TC) sponge that slowly releases tetracycline, for future periodontal applications. MATERIAL AND METHODS: Chitosan sponge was made by freezing and drying 2.5% chitosan solution. Tripolyphosphate-chitosan cross-linked (TPP) sponge was made by immersing the chitosan sponge in tripolyphosphate solution and air drying it. Tetracycline-containing chitosan (TC) sponge was prepared by freezing and drying a mixture of chitosan and tetracycline. TPP-TC sponge was made by immersing the TC sponge in tripolyphosphate solution. The weight, thickness and diameter of the four chitosan sponges were recorded. Their surface microstructures were inspected using scanning electron microscopy. The amount of tetracycline released from the sponges was analyzed by spectrophotometry. Antimicrobial activities of the residual sponges were tested against Staphylococcus aureus and Escherichia coli. RESULTS: The topography of the scaffolds was intact after the addition of tetracycline. However, increased surface irregularities were noted. In sponges with tripolyphosphate, intensified surface folding was observed. The weight of the sponges increased after tripolyphosphate and tetracycline were added, but their thicknesses and diameters decreased after cross-linking. Tetracycline was detected in the solution containing TPP-TC sponges until day 11. On day 7, the tetracycline released from TC sponges was less than that released from TPP-TC sponges. Bacterial growth was inhibited by sponges containing tetracycline. The inhibitory effect of the TPP-TC sponges was detectable until day 11. CONCLUSION: Our data showed that TPP-TC sponge was suitable as a slow-release device for tetracycline and that it maintained antimicrobial effects against the bacteria tested for up to 11 d.

The DRD2 TaqI-B polymorphism and its relationship to smoking abstinence and withdrawal symptoms.

The dopamine receptor D2 (DRD2) gene has polymorphisms that have been linked to regulation of the dopamine system and to an increased prevalence of smoking. The present study examined the relationship of the DRD2 TaqI-A and -B polymorphisms with short-term clinical outcome (abstinence and withdrawal symptoms), collected from daily (14 pre-quit and 42 post-quit) diary data among smokers (n=116) treated with the nicotine patch plus either venlafaxine or placebo. The results showed that B1/B1 or B1/B2 smokers were slightly less likely to be abstinent on a given day than those homozygous for the TaqI-B2 allele. Significant DRD2 TaqI-B x time interactions were found for several of the withdrawal scales, indicating that those smokers with the B1/B1 or B1/B2 genotypes tended to report more symptoms over time compared to those with the B2/B2 genotype. No interactions or main effects were found for the DRD2 TaqI-A polymorphism. The findings demonstrate that smokers homozygous for the TaqI-B2 allele experience progressive improvement in self-reported withdrawal symptoms while smokers with the TaqI-B1 allele showing little change.