Network-based approach for drug repurposing against mpox.

The current outbreak of mpox presents a significant threat to the global community. However, the lack of mpox-specific drugs necessitates the identification of additional candidates for clinical trials. In this study, a network medicine framework was used to investigate poxviruses-human interactions to identify potential drugs effective against the mpox virus (MPXV). The results indicated that poxviruses preferentially target hubs on the human interactome, and that these virally-targeted proteins (VTPs) tend to aggregate together within specific modules. Comorbidity analysis revealed that mpox is closely related to immune system diseases. Based on predicted drug-target interactions, 268 drugs were identified using the network proximity approach, among which 23 drugs displaying the least side-effects and significant proximity to MPXV were selected as the final candidates. Lastly, specific drugs were explored based on VTPs, differentially expressed proteins, and intermediate nodes, corresponding to different categories. These findings provide novel insights that can contribute to a deeper understanding of the pathogenesis of MPXV and development of ready-to-use treatment strategies based on drug repurposing.

Spatial Distribution Characteristics and Influencing Factors of Rural Governance Demonstration Villages in China.

Rural governance plays a significant role in constructing national governance systems and promoting rural development. An accurate understanding of the spatial distribution characteristics and influencing factors of rural governance demonstration villages is conducive to giving full play to their leading, demonstration and radiating roles and further promoting the modernization of rural governance systems and governance capacity. Therefore, this study uses Moran's I analysis, local correlation analysis, kernel density analysis and a geographic concentration index to analyze the spatial distribution characteristics of rural governance demonstration villages. Moreover, this study proposes a conceptual framework to construct the cognition of rural governance and uses Geodetector and vector data buffer analysis methods to explore the internal influence mechanism of their spatial distribution. The results show the following: (1) The spatial distribution of rural governance demonstration villages in China is unbalanced. The distribution difference between the two sides of the "Hu line" is significant. The peak appears at 30° N and 118° E. (2) The rural governance demonstration villages in China are clustered, which forms a high-density core area, a sub-high density belt, two sub-high-density centers and several single core concentration areas. Additionally, the hot spots of rural governance demonstration villages in China are mostly located on the eastern coast, tending to cluster in places with superior natural conditions, convenient transportation, and excellent economic development. (3) Based on the distribution characteristics of Chinese rural governance demonstration villages, this study proposes a "one core, three axes and multiple centers" spatial structure to optimize the distribution of rural governance demonstration villages. (4) A rural governance framework system consists of a governance subject subsystem and influencing factor subsystem. The results of Geodetector show that under the mutual leading role of the three governance subjects, the distribution of rural governance demonstration villages in China is the result of multiple factors. Among them, nature is the basic factor, economy is the key factor, politics is the dominant factor, and demographic is the important factor. The interaction network formed by general public budget expenditure and total power of agricultural machinery affects the spatial distribution pattern of the rural governance demonstration villages in China.

Climatic Variability Caused by Topographic Barrier Prevents the Northward Spread of Invasive Ageratina adenophora.

Ageratina adenophora (Spreng.) R.M.King & H.Rob. is one of the most threatening invasive alien plants in China. Since its initial invasion into Yunnan in the 1940s, it spread rapidly northward to southern Mount Nyba in Sichuan, which lies on the eastern edge of the Qinghai-Tibet Plateau. During fieldwork, we found an interesting phenomenon: A. adenophora failed to expand northward across Mount Nyba, even after the opening of the 10 km tunnel, which could have served as a potential corridor for its spread. In this work, to explore the key factors influencing its distribution and spread patterns, we used a combination of ensemble species distribution models with the MigClim model. We found that the temperature annual range (TAR), precipitation of driest month (PDM), highway density (HW), and wind speed (WS) were the most predominant factors affecting its distribution. The north of Mount Nyba is not suitable for A. adenophora survival due to higher TAR. The spatial-temporal dynamic invasion simulation using MigClim further illustrated that the northward invasion of A. adenophora was stopped by Mount Nyba. Overall, Mount Nyba may act as a topographic barrier that causes environmental differences between its south and north sides, preventing the northward invasion of A. adenophora. However, other suitable habitats on the northern side of the mountain still face challenges because A. adenophora is likely to invade via other routes. Therefore, long-term monitoring is needed to prevent human-induced long-distance spread events.

Effect of Surgical Mask on Setup Error in Head and Neck Radiotherapy.

PURPOSE: With the widespread prevalence of Corona Virus Disease 2019 (COVID-19), cancer patients are suggested to wear a surgical mask during radiation treatment. In this study, cone beam CT (CBCT) was used to investigate the effect of surgical mask on setup errors in head and neck radiotherapy. METHODS: A total of 91 patients with head and neck tumors were selected. CBCT was performed to localize target volume after patient set up. The images obtained by CBCT before treatment were automatically registered with CT images and manually fine-tuned. The setup errors of patients in 6 directions of Vrt, Lng, Lat, Pitch, Roll and Rotation were recorded. The patients were divided into groups according to whether they wore the surgical mask, the type of immobilization mask used and the location of the isocenter. The setup errors of patients were calculated. A t-test was performed to detect whether it was statistically significant. RESULTS: In the 4 groups, the standard deviation in the directions of Lng and Pitch of the with surgical mask group were all higher than that in the without surgical mask group. In the head-neck-shoulder mask group, the mean in the Lng direction of the with surgical mask group was larger than that of the without surgical mask group. In the lateral isocenter group, the mean in the Lng and Pitch directions of the with surgical mask group were larger than that of the without surgical mask group. The t-test results showed that there was significant difference in the setup error between the 2 groups (p = 0.043 and p = 0.013, respectively) only in the Lng and Pitch directions of the head-neck-shoulder mask group. In addition, the setup error of 6 patients with immobilization open masks exhibited no distinguished difference from that of the patients with regular immobilization masks. CONCLUSION: In the head and neck radiotherapy patients, the setup error was affected by wearing surgical mask. It is recommended that the immobilization open mask should be used when the patient cannot finish the whole treatment with a surgical mask.

Study on the Effect of 4D-CT Special Reconstruction Images for Evaluation of the Cardiac Structure Dose in Radiotherapy for Breast Cancer.

Objective: To study the dosimetric effect on special reconstruction images obtained from an electrocardiograph-gated four-dimensional computed tomography (ECG 4D-CT) series and compare it with the accumulation dose assessment of ECG 4D-CT. Methods: Fifteen patients underwent ECG 4D-CT scans to obtain a 4D-CT series. The 20 phase images of 0-95% were reconstructed at intervals of 5% of the cardiac cycle by the 4D-CT series. The 4D-CT series was specially reconstructed, and the maximum intensity projection (MIP), minimum intensity projection (MinIP), average intensity projection (AIP), and sum intensity projection (SIP) were obtained. The left ventricular muscle (LV) and the anterior descending branch of the left coronary artery (LAD) were delineated on all series. The intensity-modulated radiation therapy (IMRT) plan for left breast cancer was designed on the basis of the 0% phase, and the accumulative dose (Dose-acc) of 20 phases was obtained by deformation registration. The dose-volume indexes of the LV and LAD were compared based on different CT series. Results: The dose-volume indices of V5, V30, V40, Dmax, and Dmean of the LV on MIP images were 3.8, 2.0, 0.9, 3.8, and 1.7%, respectively (relative to the Dose-acc). There was no significant difference in V5 or Dmax between the MIP and Dose-acc (P > 0.05). The change rates of Dmax on the MinIP, SIP, and AIP images were 2.5, 3.1, and 1.5%, respectively (relative to the Dose-acc) (P < 0.05). Conclusion: In the dose-volume evaluation of the LV, V30, V40, and Dmean obtained by MIP were essentially the same as those obtained by the Dose-acc and can be used instead of the 4D-CT series to evaluate dose-volume indexes.

Dose accumulation in IMRT for left breast cancer determined by applying deformation registration.

BACKGROUND: To evaluate the dosimetric changes in the target volume and organs at risk (OARs) of patients with left breast cancer (LBC) who underwent intensity-modulated radiation therapy (IMRT) based on a deformation registration (DF) method. METHODS: Sixteen patients with LBC treated with 6 MV X-ray IMRT were retrospectively analyzed. All targets included the lymph node drainage area and chest wall. All patients underwent CT for simulation of the primary positioning and repositioning. Primary and secondary treatment plans were developed using primary positioning CT (CT1) and repositioning CT (CT2) images to obtain plan1 and plan2, respectively. Rigid and DF of the dose distribution of plan2 to CT1 were applied; the results were then added to the dose distribution of plan1, yielding planrig and plandef, respectively. The dosimetric differences between the target and OAR volumes of the four plans were compared. RESULTS: The clinical target volume of CT2 was 8.74% less than that of CT1. The planned target volume of CT2 was 11.20% less than that of CT1. The Dice similarity coefficients (DSCs) of the heart, left lung and right lung were significantly lower after than before DF (0.94±0.01 vs. 0.89±0.05, 0.96±0.01 vs. 0.91±0.03, and 0.96±0.01 vs. 0.92±0.03, respectively; t=3.83, 7.28, and 6.70, P<0.05, respectively). There were no significant differences in the dose-volume indices of the heart or left lung between plan1 and the other plans, while the dose-volume indices were higher in planrig than in plandef. CONCLUSIONS: Because of small changes in the target and OAR volumes during radiotherapy, we suggest the first IMRT plan could be used to evaluate the dose-volume indices of the lungs and heart for these patients.

Prevalence, risk factors, and prognosis of orthostatic hypotension in diabetic patients: A systematic review and meta-analysis.

BACKGROUND: Orthostatic hypotension (OH) is a major clinical sign of cardiovascular autonomic dysfunction in diabetic patients. Our aim was to quantitatively evaluate the prevalence and risk factors of OH in patients with diabetes mellitus (DM) and assess its prognosis. METHODS: A comprehensive search of the PubMed, Embase, China National Knowledge Infrastructure, VIP Chinese Journal, Wanfang, and SINOMED databases was conducted for related published work up to September 25, 2016, and manually searched eligible studies from the references in accordance with the inclusion criteria. RESULTS: We included 21 studies in the analysis, with a total sample size of 13,772. The pooled prevalence of OH in DM was 24% (95% confidence interval [CI]: 19-28%). Potential risk factors, that is, glycosylated hemoglobin A (HbA1c) (odds ratio [OR], 1.13, 95% CI, 1.07-1.20), hypertension (OR, 1.02, 95% CI, 1.01-1.02), and diabetic nephropathy (OR, 2.37, 95% CI, 1.76-3.19), were significantly associated with OH in DM. In addition, the prognosis of OH in DM was associated with higher risk of total mortality and cardiovascular events. CONCLUSION: The pooled prevalence of OH in DM appears high. HbA1c, hypertension, and diabetic nephropathy are risk factors for OH in DM. OH indicates poor prognosis in diabetic patients. Attention should be focused on diabetic patients with the stated risk factors to prevent OH.

BLCAP arrests G1/S checkpoint and induces apoptosis through downregulation of pRb1 in HeLa cells.

BLCAP (bladder cancer-associated protein) gene exhibited tumor suppressor function in different tumors and is regarded as a candidate tumor suppressor gene; however, the mechanism by which BLCAP exerts its function remains elusive. This study investigated the functional association between BLCAP and proliferation or apoptosis in cervical cancer cells, to identify the functional motifs of BLCAP. The BLCAP-shRNA expression vector based on pRNA-U6.1/Hygro plasmid was used to specifically inhibit BLCAP activity in HeLa cells. The optimal shRNA plasmid was selected to knock down BLCAP expression and the biological effects were investigated. The effects on cell cycle and apoptosis were detected by flow cytometric or Annexin V-FITC staining analysis. The gene expression profiles of HeLa cells transfected with blcap-wt and BLCAP-shRNA were analyzed using human signal pathway gene Oligochips. The levels of protein expression and interaction of BLCAP with Rb1 proteins were determined by western blotting and Co-IP assays. The site-specific mutagenesis assay was used to identify amino acid residues important for BLCAP. Significantly differentially expressed genes were found by gene Oligo chips analysis. These genes were all correlated with proliferation, cell cycle and apoptosis. The results of western blotting and Co-IP assays confirmed that overexpression of BLCAP could interact with Rb1 and inhibit Rb1 phosphorylation. Further investigation revealed that SAXX mutation in the key regions of BLCAP suppressed the function of BLCAP and significantly increased the level of phosphorylated Rb1 protein. Here our findings suggested that the functional association of BLCAP and Rb1 might play important roles in proliferation and apoptosis of HeLa cells. It suggested that BLCAP could be a novel therapeutic target for cervical cancer.

[Analysis of risk factors for bleeding after thyroid surgery].

OBJECTIVE: To identify risk factors for bleeding after thyroid surgery, and discuss the potential relevance between the bleeding timing and sources. METHODS: A total of 2568 cases that underwent thyroid operation from June 2012 to June 2013 were collected and analysed retrospectively the risk factors for postoperative bleeding by Cox and the potential relevance between the bleeding timing and sources. RESULTS: Among 2568 patients, 40 patients occurred postoperative bleeding Indentified risk factors were extent resection (P=0.0435) and surgeon (P=0.0071). Thyroid bed and strap muscles/sternocleidomastoid were the most common sources of bleeding after surgery within 6 hours; while thyroid bed was the most common source of bleeding after surgery between 6 and 8 h; wound errhysis was the most common source during 8-24 h after the operation; thyroid bed and strap muscles/sternocleidomastoid were the most common sources within 24 h after thgroid surgery. CONCLUSIONS: Bleeding after thyroid surgery is a rare but potentially life-threatening complication, and the extent of resection and the surgeon are risk factors. Thyroid bed and strap muscles/sternocleidomastoid are the most common sources of bleeding after surgery within 24 hours.

ABCC2 polymorphisms and survival in the Princess Margaret cohort study and the NCIC clinical trials group BR.24 trial of platinum-treated advanced stage non-small cell lung cancer patients.

BACKGROUND: The drug transporter ABCC2 is upregulated in non-small cell lung cancer (NSCLC) and implicated in platinum resistance. We evaluated the association between germline polymorphisms in the ABCC2 gene and survival outcomes of platinum-treated advanced NSCLC patients. MATERIAL AND METHODS: Ten candidate and tagging germline polymorphisms in the ABCC2 gene were genotyped in a discovery cohort of 170 platinum-treated stage IV NSCLC patients from the Princess Margaret Cancer Centre. Associations with overall survival were assessed using multivariate Cox proportional hazard models adjusted for prognostic variables. To validate our results, we analyzed the association of the two top polymorphisms in the ABCC2 gene on survival outcomes of 219 stage IIIB-IV NSCLC patients enrolled on the NCIC Clinical Trials Group BR.24 clinical trial. RESULTS: Only one polymorphism was validated across both cohorts for an association with overall survival: the A allele of the ABCC2 polymorphism, rs8187710 (4544G>A), was associated with adverse overall survival (adjusted hazard ratio [aHR] 2.22; 95% CI: 1.2-4.0; p=0.009) among our stage IV NSCLC patients. A significant association with overall survival (aHR 1.73; 95% CI: 1.0-2.9; p=0.036) was observed for the same ABCC2 polymorphism in the BR.24 validation cohort. No other ABCC2 polymorphisms were associated with outcome. CONCLUSION: The ABCC2 polymorphism, rs8187710 (4544G>A), is associated with overall survival in platinum-treated advanced NSCLC patients. Additional studies are needed to evaluate the predictive versus prognostic nature of this relationship, and to explore the functional effect of this polymorphism on the pharmacokinetics of platinum drugs.

Two BRM promoter insertion polymorphisms increase the risk of early-stage upper aerodigestive tract cancers.

Brahma (BRM) has a key function in chromatin remodeling. Two germline BRM promoter insertion-deletion polymorphisms, BRM-741 and BRM-1321, have been previously associated with an increased risk of lung cancer in smokers and head and neck cancer. To further evaluate their role in cancer susceptibility particularly in early disease, we conducted a preplanned case-control study to investigate the association between the BRM promoter variants and stage I/II upper aerodigestive tract (UADT) cancers (i.e., lung, esophageal, head and neck), a group of early-stage malignancies in which molecular and genetic etiologic factors are poorly understood. The effects of various clinical factors on this association were also studied. We analyzed 562 cases of early-stage UADT cancers and 993 matched healthy controls. The double homozygous BRM promoter variants were associated with a significantly increased risk of early stage UADT cancers (adjusted odds ratio [aOR], 2.46; 95% confidence interval [CI], 1.7-3.8). This association was observed in lung (aOR, 2.61; 95% CI, 1.5-4.9) and head and neck (aOR, 2.75; 95% CI, 1.4-5.6) cancers, but not significantly in esophageal cancer (aOR, 1.66; 95% CI, 0.7-5.8). There was a nonsignificant trend for increased risk in the heterozygotes or single homozygotes. The relationship between the BRM polymorphisms and early-stage UADT cancers was independent of age, sex, smoking status, histology, and clinical stage. These findings suggest that the BRM promoter double insertion homozygotes may be associated with an increased risk of early-stage UADT cancers independent of smoking status and histology, which must be further validated in other populations.

Resistance to bleomycin in cancer cell lines is characterized by prolonged doubling time, reduced DNA damage and evasion of G2/M arrest and apoptosis.

BACKGROUND: To establish, characterize and elucidate potential mechanisms of acquired bleomycin (BLM) resistance using human cancer cell lines. Seven BLM-resistant cell lines were established by exposure to escalating BLM concentrations over a period of 16-24 months. IC50 values and cell doubling times were quantified using a real time cytotoxicity assay. COMET and γ-H2AX assays, cell cycle analysis, and apoptosis assessment further investigated the mechanisms of BLM resistance in these cell lines. RESULTS: Compared with parental cell lines, real time cytotoxicity assays revealed 7 to 49 fold increases in IC50 and a mean doubling time increase of 147 % (range 64 %-352%) in BLM-resistant sub-clones (p<0.05 for both). Higher maintenance BLM concentrations were associated with higher IC50 and increased doubling times (p<0.05). Significantly reduced DNA damage (COMET and γ-H2AX assays), G2/M arrest, and apoptosis (p<0.05 for each set of comparison) following high-dose acute BLM exposure was observed in resistant sub-clones, compared with their BLM-sensitive parental counterparts. Three weeks of BLM-free culturing resulted in a partial return to BLM sensitivity in 3/7 BLM-resistant sub-clones (p<0.05). CONCLUSION: Bleomycin resistance may be associated with reduced DNA damage after bleomycin exposure, resulting in reduced G2/M arrest, and reduced apoptosis.

Distribution of HLA-A, -B and -DRB1 genes and haplotypes in the Tujia population living in the Wufeng Region of Hubei Province, China.

BACKGROUND: The distribution of HLA alleles and haplotypes varies widely between different ethnic populations and geographic areas. Before any genetic marker can be used in a disease-associated study it is therefore essential to investigate allelic frequencies and establish a genetic database. METHODOLOGY/PRINCIPAL FINDINGS: This is the first report of HLA typing in the Tujia group using the Luminex HLA-SSO method HLA-A, -B and -DRB1 allelic distributions were determined in 124 unrelated healthy Tujia individuals, and haplotypic frequencies and linkage disequilibrium parameters were estimated using the maximum-likelihood method. In total 10 alleles were detected at the HLA-A locus, 21 alleles at the HLA-B locus and 14 alleles at the HLA-DRB1 locus. The most frequently observed alleles in the HLA-I group were HLA-A*02 (35.48%), A*11 (28.23%), A*24 (15.73%); HLA-B*40 (25.00%), B*46 (16.13%), and B*15 (15.73%). Among HLA-DRB1 alleles, high frequencies of HLA-DRB1*09 (25.81%) were observed, followed by HLA-DRB1*15 (12.9%), and DRB1*12 (10.89%). The two-locus haplotypes at the highest frequency were A*02-B*46A (8.47%), followed by A*11-B*40 (7.66%), A*02-B*40 (8.87%), A*11-B*15 (6.45%), A*02-B*15 (6.05%), B*40-DRB1*09 (9.27%) and B*46-DRB1*09 (6.45%). The most common three-locus haplotypes found in the Tujia population were A*02-B*46-DRB1*09 (4.84%) and A*02-B*40-DRB1*09 (4.03%). Fourteen two-loci haplotypes had significant linkage disequilibrium. Construction of a neighbor-joining phylogenetic tree and principal component analysis using the allelic frequencies at HLA-A was performed to compare the Tujia group and twelve other previously reported populations. The Tujia population in the Wufeng of Hubei Province had the closest genetic relationship with the central Han population, and then to the Shui, the Miao, the southern Han and the northern Han ethnic groups. CONCLUSIONS/SIGNIFICANCE: These results will become a valuable source of data for tracing population migration, planning clinical organ transplantation, carrying out HLA-linked disease-associated studies and forensic identification.

HLA-B*07 is a high risk allele for familial cervical cancer.

AIM: The purpose of this study was to investigate 50 women from eight families with familial cervical cancer in Wufeng County, Hubei Province, China, a region with a high incidence of cervical cancer. Eighty-nine healthy women, of similar age, location and ethnicity, were selected as a control group. METHODS: Blood samples were collected from both groups, and HLA-A, HLA-B, and HLA-DRB1 genotypes were profiled with the Multi-Analyte Profiling system (xMAP) (Luminex HLA-SSO) using a WAKFlow HLA typing kit. Results were analyzed with Luminex HLA typing software and showed good stability, reproducibility and specificity. RESULTS: We found several high risk alleles in women with familial cervical cancer, that associated with the highest risk being HLA-B*07 (OR = 8.7, 95% CI = 1.8-41.1). CONCLUSIONS: HLA-B*07 is a high risk allele for cervical cancer, and has strong potential for use as a molecular biomarker.

Expression and clinical significance of epidermal growth factor receptor and type 1 insulin-like growth factor receptor in nasopharyngeal carcinoma.

OBJECTIVES: Epidermal growth factor (EGF) and type 1 insulin-like growth factor (IGF-1) receptors play an important role in the growth and apoptosis of nasopharyngeal carcinoma (NPC). They were separately found to be associated with prognosis in patients with NPC. To date, their expression correlation and clinicopathologic significance have never been specifically addressed in NPC. METHODS: Seventy-five patients with NPC and 21 noncancerous nasopharyngeal epithelial samples were accrued between 1998 and 2006 in a single hospital. The expressions of EGF and IGF-1 receptors were detected by immunohistochemical staining in the 75 NPC samples and the 21 noncancerous samples. Furthermore, the messenger RNA and protein expressions were assessed by real-time quantitative polymerase chain reaction and the Western blot technique, respectively, in NPC cell lines and normal nasopharyngeal epithelial cells. RESULTS: The 5-year survival rates, assessed by the Kaplan-Meier method, were 71.4% and 66.6% in the EGF and IGF-1 receptor protein-negative groups, respectively, whereas they were only 28.6% and 33.3% in the receptor protein-positive groups. The levels of these two proteins significantly correlated with each other, and the overexpression rates of EGF and IGF-1 receptors were 65.3% and 56% in nasopharyngeal samples, respectively. Furthermore, both protein expressions were significantly higher in NPC patients with cervical lymph node or distant metastasis than in NPC patients without lymph node or distant metastasis. Recurrence more often appears in cases positive for both proteins than in cases negative for both proteins. The expression levels of the receptor messenger RNA and proteins were higher in several NPC cell lines than in normal nasopharyngeal epithelial cells. CONCLUSIONS: These findings demonstrate that both receptor proteins may play an important role in the invasion, metastasis, and recurrence of NPC. Both receptors are valuable markers for assessing the prognosis of NPC. Their expression at such high frequencies provides the basis of combined targeted therapy with specific pharmacologic inhibitors to enhance the effects of radiotherapy and chemotherapy.

[Distribution of HLA-A, -B allele and haplotype polymorphism in the Tu nationality of Hubei province in China].

OBJECTIVE: To investigate the genetic diversity in Chinese populations. And HLA-A, -B alleles and haplotypes of 190 unrelated healthy individuals of Tu nationality from Wufeng county Hubei province were identified for the associated studies of HLA gene polymorphism and disease. METHODS: The high-resolution typing methods--sequence-based typing(SBT) was used to define the most polymorphism of exons 2 and 3 of the HLA-A, -B locus alleles. The allele and haplotype frequencies were calculated by maximum likelihood estimation with Arlequin software. RESULTS: HLA-A, -B alleles were found to be in Hardy-Weinberg equilibrium(P>0.05). A total of 26 HLA-A and 41 HLA-B alleles were detected. The most frequent alleles were A*0201(0.16053), A*110101(0.14737), A*24020101(0.14211), B*4001(0.14737), B*4601(0.13947), followed by A*0207(0.08947), A*0206(0.08158), B*1301(0.07632), B*5801(0.08947), B*1501(0.09737). The frequencies of following alleles to be A*330301(0.05526), B*1502(0.05526), B*3501(0.05263) were all higher than 0.05. The extensive HLA-A-B haplotypes were observed, and the most common haplotypes were A*0202-B*4001(0.04196), A*0201-B*4601(0.03625). CONCLUSION: In the present study, we first analyzed the HLA-A, B gene typing with SBT, all of these results will be the basic and reference data for Tu race, and also will have the applications available to trace the population migration, clinical organ transplantation, disease-associated study, HLA genetic feature and forensic identification.

[Separation of 3-substituted-(R,S)-beta-alanine derivatives by high performance liquid chromatography].

3-Substituted-(R,S)-beta-alanines derivatized by 1-fluoro-2,4-dinitro-5-L-valinamide (Marfey's reagent) were successfully separated by reversed-phase high performance liquid chromatography. The separations were performed with gradient elution. The mobile phase A was acetonitrile containing 0.1% (v/v) trifluoroacetic acid and the mobile phase B was 0.1% (v/v) trifluoroacetic acid aqueous solution. Thirty-two pairs of 3-substituted-(R,S)-beta-alanine derivatives, phenyl, substituted phenyl, naphthyl, substituted pyridyl and thienyl, were separated. The mobile phase A content was changed from 35% to 75% in 20 min. All (R-L)-diastereomers were eluted prior to the (S-L) ones. Substituents with larger hydrophobic parameters (pi) gave longer retention times (tR) for their derivatives than those with smaller ones except for 3-hydroxyphenyl and 4-hydroxyphenyl substituents. The positioning of the substituents on benzene ring of beta-alanines (beta-Ala) also influenced tR and separation. 4-Subsituted-phenyl-(R,S)-beta-Ala derivatives gave longer tR and better separation than 2-substituted isomers. The enantiomer excess values of R- and S-beta-Ala were also determined.

Characterization of a new type HPV16 E7 variant isolated from cervical cancer highest incidence area in Hubei Province of China.

AIM: To investigate the variation and biological properties of HPV16 E7 isolated from cervical cancer biopsy samples from highest incidence area in HuBei province of China. METHODS: HVP16 E7 sequences isolated from the cervical cancer biopsies of 10 local patients were amplified, sequenced and compared with prototype E7 gene. Then the variant gene was cloned into different vectors to study the antigenicity, expression and immunogenicity of its protein by Western blot, immunofluorescence and genetic immunization in vitro or in vivo. RESULTS: The results showed that 7 of 10 samples had the same mutations which led to a nonsense mutation at codon 43 of E7 sequence. The truncated E7 protein could be recognized by standard E7 monoclonal antibody in Western blot and expressed in NIH3T3 cells. In the blood sera of mice immunized intramuscularly by the plasmid DNA expressing the variant E7 gene specific E7 antibodies could be detected at week 2, 3, 5 and 6 after inoculation. However, no specific lymphoproliferation after E7 protein stimulation in vitro was detected by MTT colorimetric assay in comparison to the prototype E7 protein. CONCLUSION: HPV16 E7 gene may show variation in China and the variant protein could be expressed and induce host humoral immune response, but could not elicit special cellular-immune response against it. These data might hold the key for future development of HPV16 vaccine in HuBei province of China.