[Molecular genetic characteristics of a family which coinheritance of rare-88 C>G (HBB:c.-138 C>G) ß-thalassemia mutation with α-thalassemia and review of the literature].

The molecular genetic characteristics of a family with rare -88 C>G (HBB: c.-138 C>G) ß-thalassemia gene mutation were studied using cohort study. The cohort study was conducted from June to August 2022 by Prenatal Diagnosis Center of Sanya Women and Children's Hospital Managed by Shanghai Children's Medical Center. The phenotype and genotype were analyzed by hematological cytoanalyzer, automatic electrophoretic analysis system, and next-generation sequencing (NGS). And then, Sanger sequencing was used to verify the rare gene results. The results showed that the proband, her father, her uncle and her younger male cousin had discrete microcytosis (MCV 70.1 fl, 71.9 fl, 73.1 fl and 76.6 fl, respectively) and hypochromia (MCH 21.5 pg,22.0 pg,22.6 pg and 23.5 pg, respectively), elevated hemoglobin A2 level (5.3%, 5.4%, 5.4% and 5.5%, respectively), slightly elevated or normal fetal hemoglobin (Hb F), but no anemia. The proband was identified to have co-inherited ɑ-thalassemia (Hb Westmead gene heterozygous mutation, ɑwsɑ/ɑɑ) and ß-thalassemia with a rare -88 C>G (HBB: c.-138 C>G) heterozygous mutation (ß-88 C>G/ßN). Her mother had the same α-thalassemia as the proband. Her father, her uncle and her younger male cousin had the same rare -88 C>G heterozygous mutations as the proband. While her grandmother and younger brother were not carrier of thalassemia. In conclusion, 4 cases of rare -88 C>G(HBB:c.-138 C>G) heterozygous mutation had been detected in a Chinese family. Carriers of this beta-thalassemia are clinically asymptomatic. This study enriches the knowledge of the thalassemia mutation spectrum in Chinese people and provides valuable information for genetic counseling, prenatal diagnosis, and prevention of thalassemia, providing a scientific basis for improving the quality of birth population and preventing birth defects.

[Impact of regional positive lymph node ratio of gallbladder carcinoma on prognosis].

Objective: To study the impact of regional positive lymph node ratio (LNR) on prognosis of patients with gallbladder carcinoma. Methods: The clinicopathological and survival data of 53 patients with gallbladder carcinoma who underwent radical resection with regional lymph node metastasis in Ningbo University Affiliated Lihuili Hospital from May 2012 to December 2020 were collected, and receiver operating characteristic curve (ROC) was used to determine the optimal cut-off value of LNR for predicting postoperative survival status in patients with gallbladder carcinoma. According to the critical value, the patients were divided into low LNR group and high LNR group. The clinicopathological features and prognosis of the two groups were compared. Log rank test was used for univariate analysis of prognostic factors in patients with gallbladder carcinoma, and Cox proportional hazards model was used for multivariate analysis. Results: A total of 417 regional lymph nodes were dissected in 53 patients, of which 144 lymph nodes were positive, with a positive rate of 34.5%. The optimal cut-off value of LNR for predicting postoperative survival status of patients with gallbladder carcinoma was 0.33. According to this cut-off value, patients were divided into low LNR group (LNR≤0.33, 28 cases) and high LNR group (LNR>0.33, 25 cases). The recurrence rates were 64.3% (18/28) and 88.0 % (22/25) in low LNR group and high LNR group, respectively. The median recurrence-free survival (RFS) was 8 and 7 months, respectively (P=0.032). In the low LNR group, the 1-, 3-, and 5-year survival rates were 56.2%, 38.4%, and 32.0%, respectively, and the median overall survival (OS) was 16 months. In the high LNR group, the 1-, 3-, and 5-year survival rates were 37.9%, 5.4%, and 0, respectively, and the median OS was 9 months. The postoperative survival rate of patients in the low LNR group was better than that in the high LNR group (P=0.008). Univariate analysis showed that LNR was even associated with RFS and OS in patients with gallbladder carcinoma (P<0.05). Multivariate analysis showed that LNR>0.33 was an independent risk factor for postoperative RFS (HR=1.977, 95% CI: 1.045-3.740), but not for OS (HR=1.561, 95% CI: 0.685-3.553). Conclusion: On the basis of clearing a sufficient number of regional lymph nodes, patients with gallbladder carcinoma with regional LNR>0.33 are more likely to relapse after operation, but the predictive value of LNR>0.33 OS is insufficient.

[Efficacy of in-situ full-left/full-right split liver transplantation for adult recipients using the living donor liver transplantation technique:a single-center report of 25 cases].

Objective: To evaluate the efficacy of in-situ full size split liver transplantation(fSLT) for adult recipients using the living donor liver transplantation(LDLT) technique and to compare the characteristics of the left hemiliver graft (LHG) and the right hemiliver graft(RHG)transplantation. Methods: Deceased donor and recipient data of 25 consecutive cases of fSLT at Department of Hepatopancreatobiliary Surgery, Ningbo Medical Center Lihuili Hospital from March to December 2021 was retrieved and the patients divided into two groups:LHG group and RHG group. Among the 13 donors,11 were male and 2 were female,aged (M(IQR))38(19) years(range: 25 to 56 years),with height of 168(5) cm(range:160 to 175 cm) and weight of 65(9) kg(range: 50 to 75 kg). The median age of the 25 recipients was 52(14) years(range:35 to 71 years),17 were male and 8 were female,15 had primary liver cancer and 10 had benign end-stage liver disease,model for end-stage liver disease score was 10(9) points(range:7 to 23 points). Of the 25 recipients,10 recipients had previously undergone hepatobiliary surgery. The follow-up period was to January 2022. Demographic,clinicopathological,surgical outcomes and postoperative complications were evaluated and compared between the two groups. Continuous quantitative data were compared using Mann-Whitney U test. Classification data were expressed as frequencies,and were compared between groups using χ2 test or Fisher exact probability method. Results: Using LDLT technique,in-situ full-left/full-right liver splitting was performed and 13 viable pairs of hemiliver grafts were harvested with acquisition time of 230(53) minutes(range:125 to 352 minutes) and blood loss of 250(100) ml(range:150 to 1 000 ml). A total of 25 hemiliver grafts(13 LHG and 12 RHG) were allocated to patients listed for liver transplantation in our center by China Organ Transplant Response System. In the LHG group(13 cases),there were more females and more patients with benign end-stage liver disease than in the RHG group(12 cases)(P<0.05). The body weight and graft weight of recipients in the LHG group were lower than that in RHG group(both P<0.05). There were no significant differences in other baseline data between the two groups(all P>0.05). The graft to recipient weight ratio(GRWR) was 1.2(0.4)%(range:0.7% to 1.9%) for 25 recipients,1.1(0.5)%(range:0.7% to 1.6%)for the LHG group and 1.3(0.5)%(range:0.9% to 1.9%)for the RHG group. There was no significant difference between the two groups (P>0.05). Sharing patterns of hepatic vessels and the common bile duct are as follows:all the trunk of middle hepatic vein were allocated to the LHG group. The proportion of celiac trunk,main portal vein and common bile duct assigned to LHG and RHG was 10∶3 (P=0.009), 9∶4 (P>0.05) and 4∶9 (P=0.027),respectively. The vena cava of 12 donors in early stage retained in LHG and that of last one was shared between LHG and RHG (P<0.01). The median cold ischemia time of 25 hemiliver grafts was 240(90) minutes(range:138 to 420 minutes). For the total of 25 fSLT,the median anhepatic phase was 50(16) minutes(range:31 to 98 minutes) and the operation time was 474(138)minutes(range:294 to 680 minutes) with blood loss of 800(640) ml(range:200 to 5 000 ml). There were no significant differences in all of operation data between two groups. In the LHG group,3 patients with GRWR≤0.8% had postoperative small-for-size syndrome which improved after treatment. Postoperative Clavien-Dindo grade≥Ⅲ complications were observed in 6 cases(24.0%),4 cases(4/13) in the LHG group and 2 cases(2/12) in the RHG group,respectively. The difference was not statistically significant. Among them,5 cases improved after re-operation and intervention,1 case in LHG group died of secondary infection 2 weeks after operation,and the mortality was 4.0%. Analysis of serious postoperative complications and death has suggested that conventional caval interposition should not be used for LHG transplantation. Conclusion: Relying on accurate donor-recipient evaluation and the apply of LDLT technique,the morbidity and mortality of in-situ fSLT in adults is acceptable.

[Evaluations of total hilar en bloc resection and reconstruction in comparison with portal vein resection and reconstruction in treatment of perihilar cholangiocarcinoma].

Objective: To evaluate the feasibility and efficacy of total hilar en bloc resection and reconstruction(THERR) and portal vein resection and reconstruction(PVRR) in treatment of perihilar cholangiocarcinoma(PHC). Methods: Data of a total of 101 consecutive patients with PHC who underwent bile duct resection with various types of hepatectomies from June 2013 to December 2019 at Department of Hepatopancreatobiliary,Lihuili Hospital were retrospectively analyzed. Patients who underwent PHC resection combined with THERR or PVRR were identified and grouped accordingly. Fourteen patients(6 males, 8 females, aged (64.3±9.7)years old) underwent hepatectomy combined with THERR, 19 patients(11 males, 8 females, aged (63.8±8.6)years old) underwent hepatectomy combined with PVRR. Indications and surgical procedures of THERR and PVRR were reported. The clinicopathological characteristics and operation data, as well as the short and long-term outcomes of patients of the two groups were compared by Student's t-test and the χ2 test or Fisher exact test, respectively. The actual survivals rates were calculated by using the Kaplan-Meier method, and compared using the Log-rank test. Results: There were no statistically significant differences between the two groups in respect to age,sex and whether they had preoperative biliary drainage or not. The types of combined hepatectomy carried out predominately between the two groups were statistically different with the left side being predominant in the THERR group(10/14,P=0.010) and right side in PVRR group(12/19,P=0.001). There were no significant differences between the two groups in respect to whether they received preoperative portal vein embolization,intraoperative blood loss,curative degree,number of lymph node dissections, and whether there was lymphatic metastasis or not. However, both the times of operation and continuous Pringle maneuver were statistically longer in the THERR group((586±158)minutes and (32.5±7.3)minutes)than those in the PVRR group((453±88)minutes and (12.4±3.8)minutes),respectively(t=3.087,P=0.004;t=10.325,P<0.01). One patient in the THERR group died of liver failure 9 days postoperative, the cumulative 1-, 3- and 5-year survival rates were 84.9%, 57.1% and 37.0% for the THERR group and 81.9%, 37.8% and 30.2% for the PVRR group, respectively. There was no statistically significant differences between the two groups(χ²=0.150,P=0.698). Conclusions: Compared to the role of PVRR in the treatment of PHC, THERR is a novel and technically demanding procedure that is feasible in selected patients for the treatment of advanced PHC with invasion of both the hepatic artery and portal vein. However,due to the small size of this primary study,the value of THERR needs further evaluation.

[Split-thickness scalp and allogenic acellular dermal matrix in repairing deep wounds of hands in patients with extremely extensive burns].

Objective: To explore the clinical effects of split-thickness scalp and allogenic acellular dermal matrix (ADM) in repairing deep wounds of hands in patients with extremely extensive burns. Methods: Six patients with extremely extensive burns complicated by deep burn in hands were admitted to our department from December 2014 to December 2017, including 4 males and 2 females, aged 21 to 58 years. Their total burn areas were from 85% to 95% total body surface area (TBSA), and the sum of deep-partial thickness and full-thickness wounds was larger than 50% TBSA. Under general anesthesia, deep wounds of 10 dorsal hands were repaired by combined transplantation of split-thickness scalp and allogenic ADM in 4 to 6 weeks after injury. The skin grafting range was beyond the metacarpophalangeal joint, including partial dorsal digital deep wounds. Survival of skin grafts on dorsal hands, follow-up of hand shape and functional recovery were observed and recorded. Results: Six patients were successfully treated. There was no infection on the dorsal deep wounds of 10 hands after surgery, and the grafts survived well. During follow-up of 1 to 2 years after operation, there were no recurrent tension blisters or dorsal extension deformity of the metacarpophalangeal joints in 10 hands, the shape and function of hands recovered well, and hand Carroll scores were from 90 to 99 points, with functional levels of Ⅴ and Ⅵ. Conclusions: The combined transplantation of split-thickness scalp and allogenic ADM is a good method for repairing deep hand wounds in patients with extremely extensive burns, which can alleviate the cicatrix hyperplasia and contracture of healed hand wounds, and improve the shape and function of hands.

Standardized MR Perfusion Scoring System for Evaluation of Sequential Perfusion Changes and Surgical Outcome of Moyamoya Disease.

BACKGROUND AND PURPOSE: Simple-but-precise evaluation of cerebral perfusion is crucial for the treatment of Moyamoya disease. We aimed to develop a standardized scoring system for MR perfusion suitable for Moyamoya disease evaluation and investigate the postoperative serial changes and outcome predictors. MATERIALS AND METHODS: From January 2013 to December 2016, patients diagnosed with Moyamoya disease and receiving indirect revascularization were recruited prospectively. Clinical data and serial imaging studies were analyzed. The TTP maps were standardized using cerebellar reference values. We developed a scoring system of standardized TTP maps: 14 points for each hemisphere with higher points indicating better perfusion. RESULTS: In total, 24 children (4-17 years of age, 41 hemispheres) and 20 adults (18-51 years of age, 34 hemispheres) were included. The mean preoperative TTP scores were higher in children (7.34 ± 3.90) than in adults (4.88 ± 3.24). The standardized TTP maps revealed dynamic improvement with an increase in the corresponding scores at the 1-, 3-, and 6-month postoperative follow-ups; the scores stabilized after 6 months. The mean improvement in the 6-month scores of the pediatric and adult groups was 4.15 ± 3.55 and 6.03 ± 3.04, respectively. The 6-month TTP score improvements were associated with Matsushima grades. If we took score improvement as the outcome, the preoperative TTP score was the only significant predictor in multivariable analysis. CONCLUSIONS: The standardized TTP maps and scoring system facilitated the quantification of the sequential perfusion changes during Moyamoya disease treatment. The preoperative perfusion status was the only predictor of indirect revascularization outcome.

Giant gastrointestinal stromal tumour of the stomach: a case report with surgical treatment.

We report a case of giant gastrointestinal stromal tumour of the stomach in a 71-year-old woman, with emphasis on its going through surgical resection. The physical examination and radiological findings revealed that a giant mass occupied most of the abdominal cavity. The patient underwent an en-block resection of the mass, partial resection of the distal stomach and Billroth II gastrojejunostomy. The pathological diagnosis was gastrointestinal stromal tumour. The patient had a long-term disease-free survival. We emphasise that complete surgical resection is the only effective radical treatment approach for giant gastrointestinal stroma of the stomach. In some cases, we still have the opportunity for resection of these tumours because of their expansive growth, even though the lesions are very large.

Magnetism-Driven Ferroelectricity in Double Perovskite Y2NiMnO6.

We report the discovery of multiferroic behavior in double perovskite Y2NiMnO6. X-ray diffraction shows that the material has a centrosymmetric crystal structure of space group P2(1)/n with Ni(2+)/Mn(4+) ordering. This result is further confirmed by aberration-corrected scanning transmission electron microscopy combined with atomic resolution electron energy loss spectroscopy. The appearance of ferroelectric polarization coincides with the magnetic phase transition (∼67 K), which indicates that the ferroelectricity is driven by magnetism, and this is further confirmed by its strong magnetoelectric (ME) effect. We proposed the origin of the ferroelectricity is associated with the combination of Ni(2+)/Mn(4+) charge ordering and the ↑↑↓↓ spin ordering. When compared with other known magnetic multiferroics, Y2NiMnO6 displays several attractive multiferroic properties, including high polarization (∼145 µC/m(2)), a high multiferroic transition temperature (∼67 K), and strong ME coupling (∼21%).

Protogenin prevents premature apoptosis of rostral cephalic neural crest cells by activating the α5ß1-integrin.

The bones and connective tissues of the murine jaws and skull are partly derived from cephalic neural crest cells (CNCCs). Here, we report that mice deficient of protogenin (Prtg) protein, an immunoglobulin domain-containing receptor expressed in the developing nervous system, have impairments of the palatine and skull. Data from lineage tracing experiments, expression patterns of neural crest cell (NCC) marker genes and detection of apoptotic cells indicate that the malformation of bones in Prtg-deficient mice is due to increased apoptosis of rostral CNCCs (R-CNCCs). Using a yeast two-hybrid screening, we found that Prtg interacts with Radil, a protein previously shown to affect the migration and survival of NCCs in zebrafish with unknown mechanism. Overexpression of Prtg induces translocation of Radil from cytoplasm to cell membrane in cultured AD293 cells. In addition, overexpression of Prtg and Radil activates α5ß1-integrins to high-affinity conformational forms, which is further enhanced by the addition of Prtg ligand ERdj3 into cultured cells. Blockage of Radil by RNA interference abolishes the effect of ERdj3 and Prtg on the α5ß1-integrin, suggesting that Radil acts downstream of Prtg. Prtg-deficient R-CNCCs display fewer activated α5ß1-integrins in embryos, and these cells show reduced migratory ability in in vitro transwell assay. These results suggest that the inside-out activation of the α5ß1-integrin mediated by ERdj3/Prtg/Radil signaling is crucial for proper functions of R-CNCCs, and the deficiency of this pathway causes premature apoptosis of a subset of R-CNCCs and malformation of craniofacial structures.

Reclamation of the wastewater from an industrial park using hollow-fibre and spiral-wound membranes: 50 m3 d(-1) pilot testing and cost evaluation.

The feasibility of reclaiming effluent from industrial park wastewater treatment plants through a membrane process was evaluated in three phases. In phase 1 we selected nine wastewater treatment plants (WWTPs), each with a design capacity exceeding 10,000 m3 d(-1), and analyzed the corresponding effluent composition. 'Potential recycling percentage', R, ranged from 50% to 80% for the industrial park WWTPs, indicating a high feasibility for the reuse of effluent. In phase 2, a 50 m3 d(-1) pilot plant was installed in one of the selected WWTPs and underwent testing for one year. The quality of the reclaimed water was suitable for general-purpose industrial use. In the two ultrafiltration (UF) modules tested, the hydrophilic polyethersulfone hollow-fibre module was more tolerant to variable properties, and had higher recycling percentages than those of backwashable hydrophobic polyvinylidene difluoride spiral-wound module. Using the spiral-wound UF module helped reduce the cost for producing 1 m3 of reclaimed water (US$0.80) compared with a hollow-fibre module (US$0.88). In phase 3, we evaluated the negative effects of refluxing the reverse osmosis retentate, containing high total dissolved solids and non-biodegradable organics, with the biological treatment unit of the upstream WWTP. Biological compactibility tests showed that the refluxed retentate ratio should be reduced to maintain the conductivity of mixed liquor in the aeration tank at less than 110% of the original value.

Interleukin-1beta is increased in the cerebrospinal fluid of patients with small infarcts.

BACKGROUND AND PURPOSE: Interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) are involved in inflammatory responses during large vessel occlusion in animal models. The aim of this study was to investigate the intrathecal levels of cytokines in patients with acute small infarcts. METHODS: Forty patients with acute minor stroke and 32 non-stroke patients (including 29 age- and gender-matched subjects) who received operations with spinal anesthesia were studied prospectively and underwent measurements of cerebrospinal fluid (CSF) IL-1beta and IL-6 levels. RESULTS: After an age- and gender-matched analysis of 58 patients (29 pairs), the mean intrathecal levels of IL-1beta were 0.80 pg/ml in patients with small infarcts and 0.59 pg/ml in non-stroke patients (P < 0.0001). In addition, the mean CSF levels of IL-6 were 21.54 pg/ml and 7.52 pg/ml in the stroke and control groups, respectively (P = 0.38). These results were consistent with the data without matching. The CSF levels of IL-1beta in the 40 stroke patients were significantly higher than in the 32 non-stroke controls (P < 0.0001). CONCLUSIONS: The proinflammatory cytokine IL-1beta, but not IL-6, remained elevated in the CSF of patients in the acute stage of small infarcts.

Non-random distribution and sensory functions of primary cilia in vascular smooth muscle cells.

Although primary cilia are increasingly recognized to play sensory roles in several cellular systems, their role in vascular smooth muscle cells (VSMCs) has not been defined. We examined in situ position/orientation of primary cilia and ciliary proteins in VSMCs and tested the hypothesis that primary cilia of VSMCs exert sensory functions. By immunofluorescence and electron microscopic imaging, primary cilia of VSMCs were positioned with their long axis aligned at 58.3 degrees angle in relation to the cross-sectional plane of the artery, projecting into the extracellular matrix (ECM). Polycystin-1, polycystin-2 and alpha 3- and beta1-integrins are present in cilia. In scratch wound experiments, the majority of cilia were repositioned to the cell-wound interface. Such repositioning was largely abolished by a beta1-integrin blocker. Moreover, compared to non-ciliated/deciliated cells, ciliated VSMCs showed more efficient migration in wound repair. Lastly, when directly stimulated with collagen (an ECM component and cognate ligand for alpha 3beta1-integrins) or induced ciliary deflection, VSMCs responded with a rise in [Ca(2+)](i) that is dependent on the presence of cilia. Taken together, primary cilia of VSMCs are preferentially oriented, possess proteins critical for cell-ECM interaction and mechanosensing and respond to ECM protein and mechanical stimulations. These observations suggest a role for primary cilia in mechanochemical sensing in vasculature.

How long can patients with mild or moderate Alzheimer's dementia maintain both the cognition and the therapy of cholinesterase inhibitors: a national population-based study.

The aims of this study were to evaluate the duration of acetylcholinesterase inhibitors (AChEI) utilization as well as the patients' cognition maintenance. This study was using panel data from the Bureau of National Health Insurance (BNHI) of Taiwan from 2001 to 2004. Patients with mild or moderate AD were prescribed AChEI (donepezil, rivastigmine, or galantamine). By the regulation of BNHI, if the score of Mini-Mental Status Examination worsened by more than two points or clinical dementia rating (CDR) worsened by one or more grades in the follow-up every half year, the AChEI treatment would be terminated. Kaplan-Meier product-limit method was used to estimate duration of drug utilization. Regression model was performed to analyse the factors affecting the discontinuation of AChEI treatment. Our results showed female are more and younger than male in mild to moderate Alzheimer's dementia. The mean duration of use of AChEI was 432 days. Only 9.6% of patients maintained stable cognition tests results with continued drug refill for more than 3 years. Discontinuation rate in older patients (age > or =76 years) was higher than those in younger age (P = 0.0009). The average duration for AChEI therapy is around 14 months. The elderly are at high risk for treatment discontinuation.

Semicontinuous microcosm study of aerobic cometabolism of trichloroethylene using toluene.

A semicontinuous slurry-microcosm method was applied to mimic trichloroethylene (TCE) cometabolic biodegradation field results at the Que-Jen in-situ pilot study. The microcosm study confirmed the process of aerobic cometabolism of TCE using toluene as the primary substrate. Based on the nucleotide sequence of 16S rRNA genes, the toluene-oxidizing bacteria in microcosms were identified, i.e. Ralstonia sp. P-10 and Pseudomonasputida. The first-order constant of TCE-degradation rate was 0.5 day(-1) for both Ralstonia sp. P-10 and P.putida. The TCE cometabolic-biodegradation efficiency measured from the slurry microcosms was 46%, which appeared pessimistic compared to over 90% observed from the in-situ pilot study. The difference in the TCE cometabolic-biodegradation efficiency was likely due to the reactor configurations and the effective time duration of toluene presence in laboratory microcosms (1 days) versus in-situ pilot study (3 days). The results of microcosm experiments using different toluene-injection schedules supported the hypothesis. With a given amount of toluene injection, it is recommended to maximize the effective time duration of toluene presence in reactor design for TCE cometabolic degradation.

[Development of a rapid subtype-screening assay for the env region of HIV-1 CRF strains in Guangxi.].

BACKGROUND: To develop a simple and rapid subtype-screening assay for the env region of the circulating recombinant form (CRF) of human immunodeficiency virus type 1 (HIV-1) in Guangxi. METHODS: Proviral DNA from HIV-1 positive samples were extracted and subjected to the first round PCR with universal primers for the env region that can detect HIV-1 M group isolates. In the second round PCR, two pairs of subtype-specific primers that were designed to detect subtype C or B'/C and CRF01-AE respectively were added into one tube. The PCR products of different subtypes could be distinguished in agarose-gel electrophoresis. Additionally, all of these samples were sequenced and analyzed phylogenetically. RESULTS: Phylogenetic analysis of the env region of 50 samples showed that 3 samples (6%) were infected with CRF08-BC, 43 (86%) with CRF01-AE, and 4 (8%) remained unclassifiable. Detection of the subtype-specific primer sets revealed that 3 were subtype C or B'/C (100%), 39 were CRF01-AE (90.7%), with an adequate sensitivity (91.3%) and a high specificity (100%). Non-specific bands occasionally appeared but did not interfere with interpretation of the results. The phylogenetic analysis was consistent with subtype-specific primer sets and the consistency rate was 92%. The average reproducibility was 100% for CRF08-BC samples and 93.8% for CRF01-AE samples. CONCLUSION: A simple, rapid and low cost assay was developed for subtype-screening of CRF01-AE in Guangxi.

DNA haplotype analysis of CAG repeat in Taiwanese Huntington's disease patients.

We studied the expanded CAG repeat and adjacent CCG repeat in 53 Huntington's disease (HD) patients and 172 unrelated normal subjects matched to the patients for ethnic origin. The range of the CAG repeat varied from 38 to 109 in the HD patients and from 10 to 29 in the control group. A significant negative correlation was found between the age at onset and the CAG expansion, with no significant influence of the adjacent CCG repeat on the age at onset by multiple regression analysis. Allelic association using CCG repeat and 2 flanking dinucleotide repeat markers within 150 kb of the HD gene revealed linkage disequilibrium for 2 of 3 markers. Haplotype analysis of 24 HD families using these markers identified 3 major haplotypes underlying 87.5% of HD chromosomes. The data suggested frequent haplotypes in the Taiwanese population on which one or more mutational events leading to the disease occurred.

Dimensional analysis for establishing the testing criteria of kinetic study with respirometry.

The kinetic study of a microbial system requires the determination of kinetic parameters under a set of operating variables. Previous researchers indicated that initial conditions, such as S0/X0 and S0/KS may influence the reliability of the parameter estimation. However, little study has been done to identify the sensitivity of system variables to the parameter estimation. This study proposes a novel dimensional analysis and identifies six dimensionless groups: mu(m)/fw, kd/fw, Yg, S0/KS, KS/S0, and 1/(fw theta(c) - 1). By incorporating the SP-moving algorithm proposed by Wu and coworkers in 2001, an algorithm was proposed in this study to perform a sensitivity analysis on the six dimensionless groups. Results of this analysis reveal that S0/X0 is more sensitive than S0/KS, as also evidenced by the fact that gross growth yield (Yg) is sensitive and affecting S0/X0. The analysis also suggests that the theta(c)-based wasting frequency (fw theta(c)) is more sensitive than the daily wasting frequency (fw). A critical minimum value of 1.3 for S0/X0 and a maximum value of 0.1 for S0/KS were suggested to establish the testing criteria for the kinetic study under the respirometric conditions.

A portable, high-speed, vacuum-outlet GC vapor analyzer employing air as carrier gas and surface acoustic wave detection.

Vacuum-outlet GC with atmospheric-pressure air as the carrier gas is implemented at outlet pressures up to 0.8 atm using a low-dead-volume polymer-coated surface acoustic wave (SAW) detector. Increases in the system outlet pressure from 0.1 to 0.8 atm lead to proportional increases in detector sensitivity and significant increases in column efficiency. The latter effect arises from the fact that optimal carrier gas velocities are lower in air than in more conventional carrier gases such as helium or hydrogen due to the smaller binary diffusion coefficients of vapors in air. A 12-m-long, 0.25-mm-i.d. tandem column ensemble consisting of 4.5-m dimethyl polysiloxane and 7.5-m trifluoropropylmethyl polysiloxane operated at an outlet pressure of 0.5 atm provides up to 4 x 10(4) theoretical plates and a peak capacity of 65 (resolution, 1.5) for a 3-min isothermal analysis. At 30 degrees C, mixtures of vapors ranging in vapor pressure from 8.6 to 76 Torr are separated in this time frame. The SAW detector cell has an internal volume of < 2 microL, which allows the use of higher column outlet pressures with minimal dead time. The sensor response is linear with solute mass over at least 2-3 decades and provides detection limits of 20-50 ng for the vapors tested. The combination of atmospheric-pressure air as carrier gas, modest operating pressures, and SAW sensor detection is well-suited for field instrumentation since it eliminates the need for support gases, permits smaller, low-power pumps to be used, and provides sensitivity to a wide range of vapor analytes.

Immunogene therapy of tumors with vaccine based on Xenopus homologous vascular endothelial growth factor as a model antigen.

Overcoming immune tolerance of the growth factors associated with tumor growth should be a useful approach to cancer therapy by active immunity. We used vascular endothelial growth factor (VEGF) as a model antigen to explore the feasibility of the immunogene tumor therapy with a vaccine based on a single xenogeneic homologous gene, targeting the growth factors associated with angiogenesis. To test this concept, we constructed a plasmid DNA encoding Xenopus homologous VEGF (XVEGF-p) and control vectors. We found that immunogene tumor therapy with a vaccine based on XVEGF was effective at both protective and therapeutic antitumor immunity in several tumor models in mice. VEGF-specific autoantibodies in sera of mice immunized with XVEGF-p could be found in Western blotting analysis and ELISA assay. The purified immunoglobulins were effective at the inhibition of VEGF-mediated endothelial cell proliferation in vitro, and at antitumor activity and the inhibition of angiogenesis by adoptive transfer in vivo. The elevation of VEGF in the sera of the tumor-bearing mice could be abrogated with XVEGF-p immunization. The antitumor activity and production of VEGF-specific autoantibodies, significantly elevated IgG1 and IgG2b, could be abrogated by the depletion of CD4(+) T lymphocytes. The observations may provide a vaccine strategy for cancer therapy through the induction of autoimmunity against the growth factors associated with tumor growth in a cross reaction with single xenogeneic homologous gene and may be of importance in the further exploration of the applications of other xenogeneic homologous genes identified in human and other animal genome sequence projects in cancer therapy.

A dual-adsorbent preconcentrator for a portable indoor-VOC microsensor system.

The development and testing of a miniature dual-adsorbent preconcentrator for a microsensor-based analytical system designed to determine complex volatile organic chemical (VOC) mixtures encountered in indoor working environments at low part-per-billion levels is described. Candidate adsorbents were screened for thermal-desorption bandwidth and breakthrough volume against 20 volatile organic vapors and subsets thereof as a function of several relevant variables. A glass capillary (1.1 mm i.d.) packed with 3.4 mg of Carbopack X and 1.2 mg of Carboxen 1000 provides sufficient capacity for a 1-L dry-air sample containing all 20 vapors at concentrations of 100 ppb as well as providing a composite half-height peak width of <3 s at a desorption temperature of 300 degrees C and a flow rate of 4 mL/min. Required adsorbent masses increase to 7.0 and 1.5 mg, respectively, for the same mixture at component concentrations of 1 ppm. Vapor breakthrough volumes for the Carbopack X are unaffected by humidity from 0 to 100%RH, but those for the Carboxen 1000 are significantly reduced, requiring an additional 0.9 mg of the latter to avoid premature breakthrough at the 100 ppb level. Good peak shapes, efficient chromatographic separation of preconcentrated sample mixture components, and detection limits in the low-parts-per-billion range using an integrated surface-acoustic-wave (SAW) sensor are achieved. Preconcentrator design and operating parameters are considered in terms of the vapor bed-residence times and breakthrough volumes in the context of the modified Wheeler equation.