Rapid diagnosis of Mycobacterium marinum infection using targeted nanopore sequencing: a case report.

Mycobacterium marinum (M. marinum) is a non-tuberculous mycobacterium (NTM) that can cause infectious diseases in aquatic animals and humans. Culture-based pathogen detection is the gold standard for diagnosing NTM infection. However, this method is time-consuming and has low positivity rates for fastidious organisms. Oxford Nanopore MinION sequencing is an emerging third-generation sequencing technology that can sequence DNA or RNA directly in a culture-independent manner and offers rapid microbial identification. Further benefits include low cost, short turnaround time, long read lengths, and small equipment size. Nanopore sequencing plays a crucial role in assessing drug resistance, clinical identification of microbes, and monitoring infectious diseases. Some reports on Mycobacterium tuberculosis (MTB) using nanopore sequencing have been published, however, there are few reports on NTM, such as M. marinum. Here, we report the use of nanopore sequencing for the diagnosis of M. marinum.

[Case-control study on cortical bone trajectory screw and pedicle screw internal fixation for the treatment of senile patients with lumbar tuberculosis].

OBJECTIVE: To compare clinical effect of cortical bone trajectory (CBT) screw and pedicle screw (PS) internal fixation in treating senile patients with lumbar tuberculosis. METHODS: From January 2014 to January 2017, 42 senile patients with lumbar tuberculosis were divided into CBT group and PS group, 21 patients in each group. In CBT group, there were 12 males and 9 females, aged from 64 to 81 years old with an average of (72.52±9.25) years old, T value of bone mineral density was (-2.69±0.17) g / cm3, posterior CBT screw internal fixation and anterior debridement, interbody fusion with bone grafting was performed. In PS group, there were 11 males and 10 females, aged from 63 to 85 years old with an average of (71.42±9.81) years old, T value of bone mineral density was (-2.70±0.21) g / cm3, PS internal fixation and anterior debridement, interbody fusion with bone grafting were performed. Length of posterior incision, intraoperative bleeding volume, operation time, time of bone graft fusion and complications between two groups were compared. Level of erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), segment kyphotic Cobb angle before and after operation were compared, VAS score was used to evaluate pain releasing, JOA score was applied to evaluate clinical effect. RESULTS: All patients were followed up from 12 to 21 months with an average of (15.00±3.57) months. No reoccurrence of lumbar tuberculosis and screw loosing occurred. There were statistical difference in length of incision, intraoperative bleeding volume, operation time between two groups (P<0.05). Level ofESR and CRP between two groups at 2 weeks, 1 month and 6 months after operation were improved after operation, while there were no differences between two groups (P>0.05). There were no statistical differences in complications, time of bone graft fusion and segment kyphotic Cobb angle at 1 week after operation between two groups (P>0.05). There was difference in Cobb angle at 12 months after operation (P<0.05). For VAS score, there were no difference between two groups before operation and 3 months after operation(P>0.05), but VAS score at 3 months after operation were improved after operation between two groups (P<0.05).For JOA score, there were no difference between two groups before operation, 3 and 12 months after operation, and JOA score at 3 and 12 months after operation were improved than that of before operation between two groups (P<0.05). CONCLUSION: Both of CBT screw internal fixation and PS screw internal fixation could achieve satisfying results for the treatment of elderly patients with lumbar tuberculosis. PSinternal fixation has a long fixation but great trauma. However, CBT screw internal fixation only needs to fix adjacent segments of the lesion to reduce the fixation range, which has advantages of less trauma and strong screw holding force.

Case Report: Management of Multiple Deep-Tissue Cellulitis Without Sling Removal After an Anti-incontinence Procedure in a Female With Diabetes Mellitus.

The transobturator suburethral tape procedure is emerging as a preferred surgical option in the management of stress urinary incontinence. This procedure, also called tension-free vaginal tape transobturator (TVT-O) procedure, has fewer risks of injury to the bladder, similar effectiveness, and shorter surgery duration compared with the older tension-free vaginal tape (TVT) procedure. In this study, we report the case of a female patient with type 2 diabetes mellitus who developed emergency ketoacidosis and severe cellulitis after a TVT-O procedure, which was successfully managed without sling removal and open drainage of abscesses after multi-point puncture drainage, guided by ultrasound and appropriate antibiotic administration. The patient showed appropriate urinary continence with controlled diabetes mellitus 24 months after treatment. In conclusion, cellulitis from the pelvic floor to the associated thigh after TVT-O procedure in a diabetic patient can be managed conservatively if no sling exposure is confirmed. However, these patients should be closely observed and followed up during the perioperative period, especially for synthetic sling use.

[Feasibility study of preoperative percutaneous catheter drainage in the treatment of lumbar tuberculosis with psoas abscess].

OBJECTIVE: To investigate the effect of preoperative percutaneous drainage in lumbar tuberculosis with psoas abscess, through compare the clinical efficacy of two different methods. METHODS: Thirty-six patients with lumbar tuberculosis complicated with psoas abscess in corresponding with the inclusion criteria were admitted in Zhejiang Province Traditional Chinese Medicine Hospital from January 2015 to January 2017. The patients were divided into group A and group B according to the different therapeutic methods. All patients were given the standard anti tuberculosis treatment, and percutaneous catheter drainage was used in group A after admission immediately for psoas abscess before operation, while group B with no special treatment. At 3 weeks after anti tuberculosis treatment, 2 patients were excluded in group B, which had to postpone operation because of no reduction in erythrocyte sedimentation rate(ESR) and C reactive protein(CRP). Finally 34 patients were enrolled in this study. There were 18 cases in group A, including 10 males and 8 females, aged from 24 to 73 years old with an average of(42.5±10.2) years. There were 16 cases in group B, including 9 males and 7 females, aged from 23 to 75 years old with an average of(42.3±9.8) years. All patients were treated with posterior pedicle screw fixation, anterior debridement and bone graft fusion. Anterior surgery bleeding volume, operation time, anterior incision length, postoperative anus exhaust time, VAS score, Cobb angle, ESR, CRP changes and postoperative sinus formation were compared between two groups. RESULTS: All patients were followed up for 6 to 21 months with an average of 13 months. At the last follow-up, no mixed infection, tuberculosis recurrence, pedicle screw loosening and nail withdrawal were found. The average operative time, anterior surgery bleeding volume and anterior incision length of group A were less than that of group B(P<0.05). Postoperative anal exhaust time of group A was lower than that of group B(P<0.05). Improvement degree of ESR, CRP at 3 weeks after anti tuberculosis and 1 week after operation, group A was better than group B(P<0.05), and there was no significant difference between two groups in 1 month and 6 months after operation(P>0.05). There was 1 case sinus formation in group A and 5 cases in group B, there was significant difference in sinus formation rate between two groups (P<0.05). VAS score and Cobb angle at 1 month after operation was obvious improved(P<0.05), but there was no significant difference between two groups (P>0.05). Spinal cord nerve injury was obvious improved after operation, but there was no significant difference between two groups. CONCLUSIONS: Preoperative percutaneous catheter drainage is a safe and feasible method for the treatment of lumbar tuberculosis with psoas abscess. It can increase the effect of anti tuberculosis before operation, reduce the surgical trauma and reduce the incidence of postoperative complications. It can be recommended in clinical application.

[Clinical application of modified inverted L-shape incision by extraperitoneal approach to lumbosacral tuberculosis].

OBJECTIVE: To explore the clinical effects of one-stage posterior transpedicular screw system internal fixation combined with anterior debridement and bone grafting with modified inverted L-shape incision by extraperitoneal approach in treating multiple lumbosacral tuberculosis. METHODS: The clinical data of 15 patients with multiple lumbosacral tuberculosis underwent operation from February 2008 to December 2014 were retrospectively analyzed. There were 9 males and 6 females with an average of (47.0±13.9) years old. The lesions involved L4-S1 in 12 cases, L4-S2 in 1 case, L3-S1 in 2 cases. Five cases complicated with nerve root symptoms and 2 cases with cauda equina symptoms. All patients were treated with posterior transpedicular screw system internal fixation combined with anterior L4,5, L5S1 debridement and bone grafting with modified inverted L-shape incision by extraperitoneal approach. Operation time, blood loss, incision length, first passage of gas by anus were recorded. The condition of bone fusion and focus absorption were observed by lumbar CT and MRI; and ESR and CRP were regularly rechecked. RESULTS: Fifteen patients were followed up for 18-24 months with an average of (20.0±2.73) months. All lumbosacral pain obtained improvement, and no hardware loosening, breaking, or bone graft block loosening was found. The operative time of anterior-posterior approach surgery was 210-250 min with an average of (231.0±12.1) min; the blood loss was 320-705 ml with an average of(495.0±130.3) ml; the incision length was 15-21 cm with an average of (16.4±3.4) cm, and the extended length of inverted L-shape incision was 6 to 9 cm with an average of (7.1±2.6) cm. The time of first passage of gas by anus was 14 to 40 h with an average of (24.1±7.4) h after operation. All bone graft obtained fusion at final follow-up, and spinal cord symptoms got recovery, ESR and CRP restored normal level with no recurred at 3 months after drug withdrawal. CONCLUSIONS: The treatment of multiple lumbosacral tuberculosis with posterior transpedicular screw system internal fixation combined with anterior debridement and bone fusion with modified inverted L-shape incision by extraperitoneal approach is feasible and practical. This method has advantages of little trauma, good exposure, less complications and high security.

[Mid-term outcome of surgical operation for thoracolumbar tuberculosis].

OBJECTIVE: To investigate the mid-term outcome of operation for thoracolumbar tuberculosis. METHODS : Twenty-eight patiens with thoracolumbar tuberculosis underwent one stage anterior debridement,interbody fusion with bone graft and posterior pedicle screw internal fixation treatment from July 2006 to July 2011. There were 17 males and 11 females. Total 17 patients had nerve injuries ,including 6 cases of grade B, 5 cases of grade C, 6 cases of grade D according to Frankel classification. The poisoning symptoms of tuberculosis and recovery of spinal function were observed. The bone fusion and recovery of [umbar function were evaluated. RESULTS: All the patients were followed up ,and the duration ranged from 39 to 85 months (mean 57 months). The clinical symptoms were controlled gradually, and the thoracolumbar back pain was alleviated after operation. Among the 17 patients with complications of nerve injuries, 3 patients were improved from preoperative grade B to postoperative grade D, 3 patients were improved from preoperative grade B to postopertive E, 5 patients with preoperative grade C and 6 patients with preoperative D were almostly recovered to normal after operation. According to JOA scoring system for curative effect evaluation, the excellent and good rate at the 3rd month, the 1st year, the 3rd year and the 5th year after operation were 67.86% ,82.14% ,85.71% ,89.29% and 91.30% respectively. The results at the 6th month and the 1st year had no statistical differences compared to the results at the 3rd month (P > 0.05); but the results at the 3rd year and the 5th year were better than that at 3 months after operation (P < 0.05); and the results between 3 yesrs and 5 years after operation had no statistical differences (P < O.05). The degeneration of adjacent segments were evaluated according to the California University (Universith of California at Los Angeles , UCLA) score. The degeneration rate was 53.57% (15/28) at the 3rd year after surgery, which was better than that before surgery. Twenty-three patients were followed up for 5 years ,and the degeneration rate was 86.96% (20/23) ,which was better than those of before surgery and 3 years after surgery. CONCLUSION: The surgical treatment for thoracolumbar spinal tuberculosis can achieve the thorough debridement, reconstruction of spinal stability, recovery of lumbar function and promote the functional recovery of the spinal cord, which is an effective method of treatment. However, the mid term follow-up showed that more severe degenerative changes were found in the postoperative adjacent segment.

Haplotype data of 23 Y-chromosome markers in Minnan Han Chinese and comparison with those of 12 Y-chromosome markers.

We genotyped 23 Y-STR loci (DYS576, DYS389I, DYS448, DYS389II, DYS19, DYS391, DYS481, DYS549, DYS533, DYS438, DYS437, DYS570, DYS635, DYS390, DYS439, DYS392, DYS643, DYS393, DYS458, DYS385a/b, DYS456, and GATA-H4) in a sample of 109 unrelated male Chinese people residing in Minnan area and compared the results with those from our previous study on 12 Y-STR. The haplotype diversity and the discrimination capacity of the 23 Y-STR reached 0.9903 and 0.9725, respectively, and the genetic diversity for each locus ranged from 0.321 (DYS391) to 0.955 (DYS385). Besides, we observed a strong correlation between the number of Y-STR markers and the substantial improvement of forensic parameters used to discriminate between individuals. The results indicated that these highly polymorphic Y-STR markers were useful for human identification in forensic cases and paternity tests within the Minnan Han Chinese population.

Cross-modality PET/CT and contrast-enhanced CT imaging for pancreatic cancer.

AIM: To explore the diagnostic value of the cross-modality fusion images provided by positron emission tomography/computed tomography (PET/CT) and contrast-enhanced CT (CECT) for pancreatic cancer (PC). METHODS: Data from 70 patients with pancreatic lesions who underwent CECT and PET/CT examinations at our hospital from August 2010 to October 2012 were analyzed. PET/CECT for the cross-modality image fusion was obtained using TureD software. The diagnostic efficiencies of PET/CT, CECT and PET/CECT were calculated and compared with each other using a χ(2) test. P < 0.05 was considered to indicate statistical significance. RESULTS: Of the total 70 patients, 50 had PC and 20 had benign lesions. The differences in the sensitivity, negative predictive value (NPV), and accuracy between CECT and PET/CECT in detecting PC were statistically significant (P < 0.05 for each). In 15 of the 31 patients with PC who underwent a surgical operation, peripancreatic vessel invasion was verified. The differences in the sensitivity, positive predictive value, NPV, and accuracy of CECT vs PET/CT and PET/CECT vs PET/CT in diagnosing peripancreatic vessel invasion were statistically significant (P < 0.05 for each). In 19 of the 31 patients with PC who underwent a surgical operation, regional lymph node metastasis was verified by postsurgical histology. There was no statistically significant difference among the three methods in detecting regional lymph node metastasis (P > 0.05 for each). In 17 of the 50 patients with PC confirmed by histology or clinical follow-up, distant metastasis was confirmed. The differences in the sensitivity and NPV between CECT and PET/CECT in detecting distant metastasis were statistically significant (P < 0.05 for each). CONCLUSION: Cross-modality image fusion of PET/CT and CECT is a convenient and effective method that can be used to diagnose and stage PC, compensating for the defects of PET/CT and CECT when they are conducted individually.

Mitochondrial DNA haplogroup confers genetic susceptibility to nasopharyngeal carcinoma in Chaoshanese from Guangdong, China.

Recent studies have shown association of mtDNA background with cancer development. We analyzed mitochondrial DNA (mtDNA) control region variation of 201 patients with nasopharyngeal carcinoma (NPC) and of 201 normal controls from Chaoshan Han Chinese to discern mtDNA haplogroup effect on the disease onset. Binary logistic regression analysis with adjustment for gender and age revealed that the haplogroup R9 (P = 0.011, OR = 1.91, 95% CI = 1.16-3.16), particularly its sub-haplogroup F1 (P = 0.015, OR = 2.43, 95% CI = 1.18-5.00), were associated significantly with increased NPC risk. These haplogroups were further confirmed to confer high NPC risk in males and/or individuals ≥ 40 years of age, but not in females or in subjects <40 years old. Our results indicated that mtDNA background confers genetic susceptibility to NPC in Chaoshan Han Chinese, and R9, particularly its sub-haplogroup F1, is a risk factor for NPC.

Dominant contribution of northern chinese to the paternal genetic structure of Chaoshanese in South China.

We investigated the Y chromosome of various Chinese populations to determine the patrilineal origin of the Chaoshanese population. Admixture analysis of six specific Y short tandem repeat (STR) loci in 6,292 individual samples taken from 51 populations, including Chaoshanese and Minnanese of our earlier studies, showed that over 85% of the Chaoshanese Y chromosomes were derived from the Central China Han (M (RH): 0.8614; M (BE): 1.1868 ± 0.2054), and a very small portion were from the southern aborigines. These results support a Central China Han origin of the Chaoshanese and additionally reveal that males from the Central China Han were the predominant contributor to the patrilineal genetics of the Chaoshanese. A phylogenetic tree and analysis of molecular variance signified a strong association between Y chromosomes of Chinese populations and their linguistic affiliations, revealing a coevolution of Y chromosome diversity and languages in East Asia.

Polymorphisms of HLA-A and HLA-B genes in genetic susceptibility to esophageal carcinoma in Chaoshan Han Chinese.

Esophageal carcinoma (EC) occurs at high rate in Chaoshan region of southern China. Human leukocyte antigen (HLA) polymorphism has been implicated in risk for various cancers. To investigate the impact of HLA-A and HLA-B polymorphisms on susceptibility to EC, a case-control study was conducted among 206 patients with esophageal squamous cell carcinoma and 524 controls from Chaoshan Han population. HLA-A and HLA-B polymorphisms were genotyped by polymerase chain reaction-sequence-specific primers. Genotypic association tests for dominant, recessive, and additive models, and haplotypic association were calculated using unconditional logistic regression. A*11 was identified in a recessive model as an only allele strongly associated with EC risk (odds ratios [OR]=2.10, 95% confidence interval [CI]=1.33-3.31) even after correction for multiple test. The haplotypes A*02-B*46 (OR=1.53, 95% CI=1.04-2.24) and A*11-B*51 (OR=2.29, 95% CI=1.20-4.40) showed association with increased risk for EC, whereas A*11-B*58 (OR=0.00, 95% CI=0.00-0.82) was associated with decreased risk, though the significance of these haplotypes was lost after correction. This is a first association study at genetic level identifying HLA-A and HLA-B-related variations in genetic susceptibility to EC among Chaoshan population. The variation pattern is likely to be EC-specific because it is different from that observed for nasopharyngeal carcinoma in the same study population and might, at least in part, explain the high rate of EC in this ethnic group.

STR polymorphisms of the Henan population and investigation of the Central Plains Han origin of Chaoshanese.

Allele frequencies for 15 short tandem repeat (STR) loci were obtained from a Chinese Han population in Henan province of middle China. No deviation from Hardy-Weinberg equilibrium was observed for the STR loci except for D3S1358. The 15 STR loci are potentially useful for paternity testing and forensic casework in the Henan population. A phylogenetic tree based on CODIS STR allele frequencies of 25 Han populations revealed noticeable but far less clear distinctions between southern and northern Chinese populations; the Henan Han population was located at an intermediate position between south and north Chinese Han populations, relatively closer to Chaoshan and Minnan Han. Moreover, admixture analysis showed a large proportion of Central Plains Han origin in Chaoshanese and Minnanese. Admixture and phylogenetic analysis also reflected the genetic similarity shared by these two groups.

Suppression of gastric cancer growth by baculovirus vector-mediated transfer of normal epithelial cell specific-1 gene.

AIM: To study the inhibitory effect of baculovirus-mediated normal epithelial cell specific-1 (NES1) gene therapy on gastric cancer (GC) in vitro and in vivo. METHODS: We first constructed recombinant baculovirus vectors and then transfected them into gastric cancer cells (SGC-7901). Efficiency of the baculovirus for gene transfer into SGC-7901 cells and cell growth curves were detected by fluorescence microscopy, Western blot and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro, respectively. The therapeutic effect of this gene therapy on GC was confirmed in xenografted nude mice. Tumor growth was determined by tumor volume, and expression of NES1 in tumor was analyzed by immunohistochemistry. RESULTS: Baculovirus vectors were successfully transfected into SGC-7901 cells. SGC-7901 cells transfected with the NES1 gene inhibited cell growth. In the Bac-NES1 treated group, tumor growth was significantly reduced with a high level of NES1 expression CONCLUSION: Baculovirus-mediated NES1 gene can be used in gene therapy for GC.

[Experimental study of baculovirus-mediated transfection of spiral ganglion cells in rats].

OBJECTIVE: To study the feasibility and the characteristics of recombinant baculovirus as spiral ganglion cells (SGC) gene transfer vector. METHODS: After the generation of baculovirus- green fluorescent protein( Bac-GFP) according to Bac-to-Bac baculovirus expression system, SGC were infected by Bac-GFP with different multiplicities of infection (MOI) and different concentrations of sodium butyrate. The transfection cell rate and mean fluorescence strength (MFS) were detected by fluorescence microscopy and flow cytometry. Toxicity effects of recombinant baculovirus vectors and sodium butyrate on SGC were determined by spectroscopic measurement of 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-diphenytetrazoliumromide (MTF). RESULTS: Baculovirus was able to infect primary SGC cultures. The dose-response characteristics of Bac-GFP were determined on SGC, and the expression level could be up-regulated by sodium butyrate. Infection with Bac-GFP in the absence or presence of sodium butyrate (< or =10 mmol/L) was considered to be non-cytotoxic to primary SGC. GFP had been expressed in SGC at 6 h post-infection and the highest numbers of cells expressing GFP were observed at approximately 48 h post-infection. CONCLUSIONS: Baculovirus is a novel and promising tool for gene transferring into the cochlear nervous system both for studies of the function of foreign genes and the development of gene therapy strategies.

Genetic link between Chaoshan and other Chinese Han populations: Evidence from HLA-A and HLA-B allele frequency distribution.

The genetic polymorphism of HLA-A and HLA-B loci was investigated in 505 Chaoshanese using PCR-SSP method. Among the HLA-A alleles detected, A*11 (35.64%) was most frequent, followed by A*02 (31.78%). Of 34 HLA-B alleles tested, 30 were observed, in which B*60 (21.68%), B*46 (14.46%), and B*58 (10.69%) were highly predominant. Comparison was made with other nine Chinese Han ethnic groups covering the Mainland China, Taiwan, Hong Kong, and Singapore. The high frequent alleles found in Chaoshanese were also common in other Chinese groups compared though the frequency levels varied from group to group. The phylogenic tree analysis based on the HLA-A and -B allele frequencies of all the 10 Chinese ethnic groups revealed that Chaoshanese, while clustering in general with the southern China-related Han Chinese, had the highest affinity to the Mainland Minnanese, but separated distinctively from the northern Han Chinese. The study, however, was yet to confirm the hypothesis of the Central Plains Han origin of Chaoshanese. Interestingly, the alleles (B*46, B*38, and B*58) and the related haplotypes (A*02-B*46 and A*33-B*58) that are positively associated with nasopharyngeal carcinoma (NPC), a disease prevailing predominantly among southern Chinese, were always at much higher frequencies among southern Chinese than among northern Chinese, whereas A*31 and B*13, the two alleles with highly protective effects for NPC, and the associated haplotype A*30-B*13 were predominantly high in northern Chinese. The different genetic background between northern and southern China may explain, at least partially, the prevalence of NPC among southern Chinese.

[An analysis for genetic polymorphism of twelve Y short tandem repeat loci in Chaoshan populations].

OBJECTIVE: To study the genetic polymorphism of twelve short tandem repeat loci of Y chromosome in Chaoshan Han populations. METHODS: Twelve Y short tandem repeat loci, of which the template DNAs were extracted from blood samples of 121 unrelated Chaoshan Han individuals, were amplified by using the PowerPlex Y system. The PCR products were genotyped with ABI PRISM 3100 genetic analyzer. RESULTS: From the loci DYS19, DYS437, DYS389I, DYS389II, DYS438, DYS439, DYS393, DYS391, DYS390, and DYS392, 4-7 alleles were checked out; from the loci DYS385, 35 alleles were observed. The overall observed haplotype diversity reached 0.9976. DYS391 was the locus with the lowest gene diversity (0.4445), while DYS385 had the highest gene diversity (0.9525). CONCLUSION: The 12 Y-STR loci are highly polymorphic and suitable to forensic casework, population genetics and anthropology studies.

[Manual microdissection of defined cells and RNA extraction for gene expression analysis of esophageal carcinoma progress].

OBJECTIVES: To isolate cells of interest from heterogeneous tissue blocks to obtain accurate representations of molecular alterations acquired by neoplastic cells so as to meet the demands of further study on gene expression patterns of the esophageal carcinoma (EC) evolution. METHODS: Blocks of EC were stored at -70 degrees C as close as possible to the time of surgical resection. The tissue block was embedded in OCT and frozen sections of 35 microns in thickness were cut in a cryostat under strict RNAse-free conditions. Individual frozen sections were mounted on plain glass slides and 30-gauge needle attached to a 1 ml syringe was used to microdissect defined cells in the sections. The procured cells were used for total RNA extraction. RESULTS: An optimized protocol of manual microdissection was developed successfully whereby regions with an area as small as 1/25 mm2 could be accurately dissected. The RNA recovered from procured cells was of high quality suitable for subsequent applications of molecular analysis as assessed of 18S and 28S rRNAs by electrophoresis on agarose gel. CONCLUSIONS: It is believed that manual microdissection is capable to procure defined cell populations from complex primary tissues, thus allowing investigation of tissue-, cell-, and function-specific gene expression patterns. The technique is simple, easy to perform, versatile, and of particular usefulness when laser capture microdissection (LCM) is practically unavailable.

Detection of human papillomavirus in esophageal carcinoma.

The aim of the study was to assess the prevalence of human papillomavirus (HPV) in the esophagus in the coastal region of Eastern Guangdong, Southern China, an area with a high incidence of esophageal carcinoma. Fresh surgical resection esophageal specimens were obtained from 176 esophageal carcinoma patients admitted to the Tumor Hospital of Shantou University Medical College. The samples were subjected to polymerase chain reaction (PCR) to detect HPV infection using consensus and type-specific primers for HPV type 6, 11, 16, and 18. The incidence rate was 65.5%, 69.1%, and 60% in tissues of cancerous, paracancerous and normal mucosa, respectively. Further analysis of the distribution of HPV types in the three sections of tissues showed that the high-risk HPV types 16 and 18 were found mainly in the cancer cells (43.2%), whereas the low-risk HPV types 6 and 11 were seen mainly in the normal mucosa (52.3%). The total infection rate of the high-risk HPV types 16 and HPV 18 was the highest in cancerous tissues (54.5%), followed by paracancerous tissues (19.5%), and the lowest in normal mucosa (11.7%). There was high incidence of HPV infection in the esophageal epithelium in Eastern Guangdong, Southern China, where esophageal carcinoma is prevalent. HPV was seen in the normal, paracancerous and cancerous tissues, with the high-risk HPV type 16 and 18 more common in cancerous tissues. The results indicate that the high incidence of esophageal carcinoma in this area is associated with HPV infection.