Clinicopathological characteristics of myelodysplastic syndromes with del(5q) in Taiwan.

BACKGROUND: Myelodysplastic syndromes (MDS) are a group of clonal haematopoietic stem cell disorders characterised by ineffective haematopoiesis and cytopenia. Studies have reported differences in MDS between Asian and Western countries, but data from Taiwan are scarce. MATERIALS AND METHODS: In this study we analysed the clinical and pathological features of 32 Taiwanese MDS patients with del(5q) (ie, del(5q) alone [Group A, n = 11], del(5q) with one additional cytogenetic abnormality other than monosomy 7 or del(7q) [Group B, del(5q)+1; n = 6], and del(5q) with ≥2 additional cytogenetic abnormalities [Group C, n = 15]). RESULTS: Progression-free survival (PFS) and overall survival (OS) were more favourable for Group A than for Groups B (p < 0.05) and C (p ≤ 0.001). Multivariate analysis showed that age >70 years, thrombocytopenia, and karyotype other than del(5q) alone were poor prognostic factors. Among the patients that had World Health Organization (WHO)-defined MDS with isolated del(5q), one patient (9%) had a typical marrow morphology of 5q minus syndrome with erythroid hypoplasia and four patients (36%) had hypolobated megakaryocytes. In addition, PFS and OS were significantly more favorable for the patients with del(5q) alone than for those with del(5q)+1 (p < 0.05). CONCLUSION: The bone marrow morphology, clinical features, and prognosis of Taiwanese MDS patients with del(5q) were different from those associated with MDS with isolated del(5q) as defined in the current WHO classification. Researchers should compare different geographic regions and racial populations to determine whether geographic and racial differences exist with respect to MDS with del(5q).

[Research advances on the prevention and treatment of burn infection in the elderly].

Infection is a common complication after burns and the major cause of death in patients suffering severe burn injury. The infection of the elderly after burns is more serious due to their decreased immune function that is complicated with factors such as multiple chronic diseases and dysfunction of various organs. In addition, the burn infection in the elderly lacks the specific symptoms and signs, which brings great challenges to its diagnosis and treatment. To effectively prevent and control infection is very important for the treatment of elderly burn patients. Combined the clinical characteristics of burn infection in the elderly, this paper summarized the research advances of prevention and treatment for burn infection in the elderly from fluid resuscitation, wound treatment, antibiotic using, organ protection, nutritional support, and infection prevention, aiming to provide reference for clinical practice.

Pollinators shift to nectar robbers when florivory occurs, with effects on reproductive success in Iris bulleyana (Iridaceae).

Studies have indicated that florivory and nectar robbing may reduce reproductive success of host plants. However, whether and how these effects might interact when plants are simultaneously attacked by both florivores and nectar robbers still needs further investigation. We used Iris bulleyana to detect the interactions among florivory, nectar robbing and pollination, and moreover, their effects on plant reproductive success. Field investigations and hand-pollination treatments were conducted on two experimental plots from a natural population, in which Experimental plot was protected from florivores and Control plot was not manipulated. The flower calyx was bitten by sawflies to consume the nectary, and three bumblebee species were pollinators. In addition, the short-tongued pollinator, Bombus friseanus, was the only robber when there was a hole made by a sawfly. The bumblebee had significantly shortened flower handling time when robbing, as compared to legitimate visits. Pollinator visitation and seed production decreased significantly in damaged flowers. However, seed production per flower after supplementary hand-pollination did not differ significantly between damaged and undamaged flowers. Compared to the Experimental plot, bumblebees visited fewer flowers per plant in a foraging bout in the Control plot. The flowers damaged by florivory allowed B. friseanus to shift to a nectar robber. Florivory and nectar robbing collectively decreased plant reproductive success by consuming nectar resources, which may reduce attractiveness to pollinators of the damaged flowers. However, the changes in pollinator behaviour might be beneficial to the plant by reducing the risk of geitonogamous mating.

Distribution and diversity of bacterial communities and sulphate-reducing bacteria in a paddy soil irrigated with acid mine drainage.

AIMS: To investigate the effects of long-term acid mine drainage (AMD) irrigation on the change in bacterial community and sulphate-reducing bacteria (SRB) in a paddy soil. METHODS AND RESULTS: The bacterial community structures were investigated using 454 pyrosequencing, and 98 931 effective sequences were selected for the bacterial diversity analysis. The known dominant phyla in the paddy soil were Acidobacteria (33·5%), Proteobacteria (19·7%), Nitrospira (2·8%) and Actinobacteria (2·7%). Higher percentage of Acidobacteria than Proteobacteria was detected. The relative abundances of the dominant bacterial lineages were more significantly correlated with the soil pH, the organic matter and the sulphate than the heavy metals. The diversity of the SRB in the surface paddy soil showed that the uncultured SRB groups might play important roles in paddy soils. The other OTUs mainly belonged to six phylogenetic divisions: Desulfobacca, Desulfovibrio, Syntrophobacter, Desulforhopalus, Desulfarculus and Desulfobulbus. The distribution of the absolute abundance and the relative contribution of the SRB along the vertical soil profile were investigated by RT-PCR assays based on the dsrB gene. The abundance of the dsrB gene copy numbers was up to 1·92 × 10(9)  copies g(-1) dry soil, which is slightly higher than the other non-AMD-affected paddy soil. CONCLUSIONS: This study demonstrated that the abundance of SRB is increased by the AMD irrigation while changing the composition and diversity of the bacterial community in the paddy soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This is, to our knowledge, the first attempt to characterize and quantify changes in the diversity and distribution of the microbial community and SRB in the long-term AMD-irrigated paddy soil, which will further increase our understanding of the impact of AMD on sulphur biogeochemical cycling in the paddy soil.

EB-virus latent membrane protein 1 potentiates the stemness of nasopharyngeal carcinoma via preferential activation of PI3K/AKT pathway by a positive feedback loop.

Our previous study reported that Epstein-Barr virus(EBV)-encoded latent membrane protein 1 (LMP1) could induce development of CD44(+/High) stem-like cells in nasopharyngeal carcinoma (NPC). However, the molecular mechanisms that underlie modulation of cancer stem cells (CSCs) in NPC remain unclear. Here, we show that LMP1 induced CSC-like properties through promotion of the expression of epithelial-mesenchymal transition-like cellular markers and through alterations in differentiation markers. Furthermore, LMP1 activated and triggered phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway, which subsequently stimulated expression of CSC markers, development of side population and tumor sphere formation. This suggests that PI3K/AKT pathway has an important role in the induction and maintenance of CSC properties in NPC. Similarly, PI3K/AKT pathway was also activated by phosphorylase in LMP1-induced CD44(+/High) cells. In addition, LMP1 greatly increased expression of miR-21 and downregulated expression of the miR-21 target, PTEN. Overexpression of miR-21 by transfection of miR-21 mimics into LMP1-transformed cells led to phosphorylase-mediated activation of the PI3K/AKT pathway and induction of CSCs. On the contrary, phosphorylation of the PI3K/AKT pathway and the expression of CSC were reversed by an miR-21 inhibitor. The specific inhibitor (Ly294002) of PI3K/AKT pathway significantly decreased expression of miR-21 and CSC markers and upregulated the expression of PTEN, which indicates that miR-21 and PTEN are the downstream effectors of PI3K/AKT and that expression of these two effectors are related to the development of NPC CSCs. Taken together, our novel findings indicate that LMP1, PI3K/AKT, miR-21 and PTEN constitute a positive feedback loop and have a key role in LMP1-induced CSCs in NPC.

Adult T-cell leukaemia/lymphoma can mimic other lymphomas in a non-endemic area: dilemmas in diagnosis and treatment.

BACKGROUND: The diagnosis of Adult T-cell leukaemia/lymphoma (ATL) in non-endemic regions is challenging. AIM: This study analyses the clinicopathologic features and diagnostic processes of ATL patients in Taiwan. METHODS: ATL patients diagnosed and treated at Taipei Veterans General Hospital from 1998 through 2010 were retrospectively identified. The diagnosis of ATL was confirmed by in situ detection of human T-cell leukaemia virus type 1 (HTLV-1) when necessary. Patients' data were reviewed and analysed. RESULTS: Fourteen ATL patients were identified, among whom six (42.9%) had an antecedent diagnosis of other malignant lymphomas before the ATL diagnosis, including two diagnosed with Hodgkin disease (HD), one with peripheral T-cell lymphoma, two with chronic lymphocytic leukaemia and one with angioimmunoblastic T-cell lymphoma. Of the 14 patients, eight (57%) were subclassified as the acute type, three (21.4%) as the lymphoma type, and three (21.4%) as the chronic type ATL. Five of six (83.3%) patients with initial non-ATL misdiagnosis were diagnosed with non-acute type ATL. In particular, a patient with an antecedent diagnosis of HD presented with typical Reed-Sternberg (RS)-like cells harbouring Epstein-Barr virus genomes in affected lymph nodes. The patient progressed to acute type ATL 3 years after the initial diagnosis, and HTLV-1 genomes were identified in the previous RS-like cells. CONCLUSION: In non-endemic areas, such as Taiwan, ATL, particularly the non-acute type, may mimic other lymphomas and easily be misdiagnosed. HTLV-1 serology should be routinely screened in all malignant lymphoma patients. In situ detection of HTLV-1 is helpful in cases with diagnostic dilemmas.

Pollen packaging and dispensing: adaption of patterns of anther dehiscence and flowering traits to pollination in three Epimedium species.

Pollen presentation theory (PPT) predicts that plant species typically pollinated by frequent and wasteful pollinators ought to be much more parsimonious and only gradually release pollen compared to plant species pollinated by infrequent pollinators that are efficient at delivering the pollen they remove. To test PPT, we compare the pollen presentation schedules and pollination systems in three related Epimedium species, having different pollinators. Results showed that differences in anther dehiscence and flowering traits resulted in different pollen packaging schedules. For E. sutchuenense and E. franchetii, a special 'roll-up' movement of the anther wall during anther dehiscence increased pollen removal compared to the dehiscence pattern in E. mikinorii, which lacked the 'roll-up' movement. Investigations revealed that honeybees had a higher pollen removal rate and lower stigmatic pollen load compared to bumblebees. In accordance with PPT, E. sutchuenense presents pollen sequentially and slowly for the frequent and wasteful honeybees. In comparison to E. sutchuenense, E. franchetii had a faster presentation rate and was adapted to the efficient and infrequent bumblebees. However, E. mikinorii was pollinated by both bumblebees and honeybees at high frequency and had the fastest pollen presentation. This pattern could reduce pollen wastage by honeybees and might be an adaptation to its short flower longevity (less than 1 day), to increase the chances of pollen deposition on stigmas. The study indicates that pollen presentation schedules can be a consequence of interactions among anther dehiscence, flowering traits and pollination environments for a given species.

The differential contributions of herkogamy and dichogamy as mechanisms of avoiding self-interference in four self-incompatible Epimedium species.

Self-interference is one of the most important selective forces in shaping floral evolution. Herkogamy and dichogamy both can achieve reductions in the extent of self-interference, but they may have different roles in minimizing self-interference in a single species. We used four self-incompatible Epimedium species to explore the roles of herkogamy and dichogamy in avoiding self-interference and to test the hypothesis that herkogamy and dichogamy may be separated and become selected preferentially in the taxa. Two species (E. franchetii and E. mikinorii) expressed strong herkogamy and weak protogyny (adichogamy), whereas another two species (E. sutchuenense and E. leptorrhizum) expressed slight herkogamy and partial protandry. Field investigations indicated that there was no physical self-interference between male function and female function regarding pollen removal and pollen deposition in all species. Self-pollination (autonomous or facilitated) was greater in species with slight herkogamy than in those with strong herkogamy. Artificial pollination treatments revealed that self-pollination could reduce outcrossed female fertility in all species, and we found evidence that self-interference reduced seed set in E. sutchuenense and E. leptorrhizum in the field, but not in E. franchetii and E. mikinorii. These results indicate that well-developed herkogamy is more effective compared with dichogamy in avoiding self-interference in the four species. In genus Epimedium, herkogamy instead of dichogamy should be selected preferentially and evolved as an effective mechanism for avoiding self-interference and might not need to evolve linked with dichogamy.

How to predict the outcome in mature T and NK cell lymphoma by currently used prognostic models?

To select an appropriate prognostic model in the treatment of mature T- and natural killer (NK) -cell lymphoma (peripheral T-cell lymphoma (PTCL) and NK-/T-cell lymphoma (NKTCL)) is crucial. This study investigated the usefulness of Ann Arbor staging classification International prognostic index (IPI), prognostic index for T-cell lymphoma (PIT) and International peripheral T-cell lymphoma Project score (IPTCLP). Between 2000 and 2009, 176 patients (122 males) with PTCL and NKTCL were diagnosed and treated from a single institute in Taiwan. The correlation between complete response (CR) rate, 3-year overall survival (OS), early mortality rate and four prognostic models was analyzed. Thirty-one patients received hematopoietic stem cell transplantation (HSCT) and were analyzed separately. Three-year OS rate was 34.7%, and anaplastic large-cell lymphoma harbored better outcome than others. IPI score had the lowest Akaike information criterion value (1081.197) and was the best score in predicting OS and early mortality (P=0.009). Ann Arbor stage classification can predict CR rate more precisely (P=0.006). OS was significantly better in patients who received HSCT, even in patients with unfavorable features compared with chemotherapy alone. All prognostic models were useful to evaluate the outcome of patients with PTCL and NKTCL but IPI score did best in predicting OS in PTCL and PIT score in NKTCL. This study also supported the role of HSCT in patients with high-risk or refractory PTCL or NKTCL.

Identification of two novel HLA-B*40 alleles, B*40:137 and B*40:158, in Taiwanese individuals.

Using sequence-based typing method we discovered two new HLA-B*40 variants, B*40:137 and B*40:158, in Taiwanese individuals. The sequence of B*40:137 has three nucleotide (nt) changes from B*40:21 at nt 353 (C→T), nt 355 (C→A) and nt 369 (C→T) resulting two coding changes at residue 94 (T→I) and residue 95 (L→I), whereas the sequence of B*40:158 differs from B*40:01:01 with five nt substitutes at nt 463 (C→A), nt 477 (C→G), nt 499 (T→A), nt 512 (T→G) and nt 527 (T→A) causing five amino acid exchanges at codons 140 (Y→S), 155 (R→S), 168 (S→T), 171 (L→W) and 179 (V→E). Our hypotheses on the generation of the two novel alleles are presented.

An HLA-A*02:01-B*13:01-DRB1*14:01:03 haplotype conserved in Taiwanese and a possible close relationship between DRB1*14:01:03 and DRB1*14:54.

We here report sequence confirmation and analysis of the variant HLA-DRB1*14:01:03 on three voluntary bone marrow donors and the conserved haplotype carrying DRB1*14:01:03 allele in Taiwanese population. In exon 2, the DNA sequence of DRB1*14:01:03 is identical to HLA-DRB1*14:01:01 except a silent nucleotide substitution at position 192. However, sequence specific primer (SSP) reaction pattern of DRB1*14:01:03 matched with the pattern of DRB1*14:54 instead of DRB1*14:01:01, 14:01:02 or 14:01:03. In exon 3, at position 421, DRB1*14:01:03 has an identical nucleotide as DRB1*14:54 but differs from DRB1*14:01:01. We think the discrepancy of the allele assignment by SSP typing protocol and by sequence-specific oligonucleotide probe (SSO) and sequence-based typing methods should be addressed. We assume DRB1*14:54 is probably the parental allele for DRB1*14:01:03.

Behavioural differences between male and female carpenter bees in nectar robbing and its effect on reproductive success in Glechoma longituba (Lamiaceae).

Male and female nectar robbers may show significantly different behaviour on host plants and thus have different impacts on reproductive fitness of the plants. A 4-year study in natural populations of Glechoma longituba has shown that male carpenter bees (Xylocopa sinensis) are responsible for most of the nectar robbing from these flowers, while female bees account for little nectar robbing, demonstrating distinct behavioural differentiation between male and female bees in visiting flowers. The smaller male bee spends less time visiting a single flower than the larger female bee, consequently, the male bee is capable of visiting more flowers per unit time and has a higher foraging efficiency. Moreover, the robbing behaviour of female carpenter bees is more destructive and affects flower structures (ovules and nectaries) and floral life-span more than that of the male bee. According to the energy trade-off hypothesis, the net energy gain for male bees during nectar robbing greatly surpasses energy payout (17.72 versus 2.43 J), while the female bee net energy gain is barely adequate to meet energy payout per unit time (3.78 versus 2.39 J). The differences in net energy gain for male and female bees per unit time in nectar robbing are the likely cause of observed behavioural differences between the sexes. The differences in food resource preference between male and female bees constitute an optimal resource allocation pattern that enables the visitors to utilise floral resources more efficiently.

Single-walled carbon nanotube based pH sensors on a flexible parylene-C substrate.

In this paper, we present a suspended Single-Walled Carbon Nanotube (SWNT) based pH sensor utilizing a low temperature Dielectrophoretic (DEP) assembly process on a flexible parylene-C substrate. Parylene-C, a light weight, flexible and inert material, is compatible with many microfabrication processes. Furthermore, utilizing parylene-C as a flexible substrate, one can readily create a suspended microplatform utilizing an O2 plasma etch process. Suspended nanobridges have larger exposed surface areas and may potentially have enhanced sensitivity for sensing applications. Fabricating these structures on a thin (10 microm) parylene-C substrate allows their utilization as flexible devices or in wearable sensor applications. We have successfully assembled suspended SWNT nanobridges across a spacing of 4 microm. The electrical characterization results from the assembled SWNTs yield ohmic behavior with a measured two-terminal resistance of approximately 17Komega. Furthermore, the conductometric measurements of the SWNT sensors have demonstrated that corresponding to an increase in pH value, the resistance of SWNTs has decreased due to the OH- group that attached on to the wall of the SWNTs and changed the electrical properties of the SWNTs. These novel suspended nanostructures can be used as potential candidates in nanosensor applications.

Hospital-based surveillance and molecular epidemiology of rotavirus infection in Taiwan, 2005-2007.

To determine the distribution of rotavirus strains and facilitate vaccine policy decisions in Taiwan, active hospital-based gastroenteritis surveillance was conducted in three sentinel hospitals. From 1 January 2005 to 31 December 2007, a total of 3435 children less than 5 years old with gastroenteritis were enrolled. The presence of rotavirus was documented by enzyme immunoassay (EIA), and the G and P genotypes were determined by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing methods. Results confirmed that 856 (25%) of these gastroenteritis admissions were EIA-positive for rotavirus and 448 (52%) of the rotavirus positive admissions were less than 2 years old. The most prevalent rotavirus genotypes were G1P[8] (40%), followed by strains G3P[8] (27%), and G9P[8] (17%). These data will help inform decisions as to whether rotavirus vaccine should be considered for inclusion into Taiwan's National Immunisation Programme.

Selective nectar robbing in a gynodioecious plant (Glechoma longituba) enhances female advantage.

Nectar robbing not only affects the reproductive fitness of the plant but it may also potentially affect the pollination dynamics of the associated coflowering individuals. In this study, we established that the nectar robber Xylocopa sinensis robs nectar only from the hermaphrodite ramets of the gynodioecious plant Glechoma longituba but not from the female ramets. In populations with high rates of nectar robbing, this results in hermaphrodite ramets having reduced seed set whereas the female ramets have a slightly increased seed set. We hypothesize that selective nectar robbing confers an advantage to female individuals and thus ensures their maintenance in gynodioecious populations. Results of controlled experiments indicated that the reduction in the amounts of nectar available occasioned by nectar robbing resulted in some legitimate pollinators switching to visiting flowers on female rather than hermaphrodite ramets. This resulted in lower pollination rates and seed set for hermaphrodites and higher pollination rates and seed set for females. This study presents a previously unreported mechanism causing female advantage in gynodioecious plants.

Genetic variation among wild and cultivated populations of the Chinese medicinal plant Coptis chinensis (Ranunculaceae).

To examine if the cultivation process has reduced the genetic variation of modern cultivars of the traditional Chinese medicinal plant, Coptis chinensis, the levels and distribution of genetic variation was investigated using ISSR markers. A total of 214 C. chinensis individuals from seven wild and three cultivated populations were included in the study. Seven ISSR primers were used and a total of 91 DNA fragments were scored. The levels of genetic diversity in cultivated populations were similar as those in wild populations (mean PPL = 65.2% versus PPL = 52.4%, mean H = 0.159 versus H = 0.153 and mean I = 0.255 versus I = 0.237), suggesting that cultivation did not seriously influence genetic variation of present-day cultivated populations. Neighbour-joining cluster analysis showed that wild populations and cultivated populations were not separated into two groups. The coefficient of genetic differentiation between a cultivar and its wild progenitor was 0.066 (G(st)), which was in good accordance with the result by amova analysis (10.9% of total genetic variation resided on the two groups), indicating that cultivated populations were not genetically differentiated from wild progenitors. For the seven wild populations, a significant genetic differentiation among populations was found using amova analysis (45.9% of total genetic variation resided among populations). A number of causes, including genetic drift and inbreeding in the small and isolated wild populations, the relative limited gene flow between wild populations (N(m) = 0.590), and high gene flow between cultivars and their wild progenitors (N(m) = 7.116), might have led to the observed genetic profiles of C. chinensis.

Prevalence of the JAK2-V617F mutation in Taiwanese patients with chronic myeloproliferative disorders.

BACKGROUND: The Janus kinase-2 (JAK-2) V617F mutation has been recently reported in patients with myeloproliferative disorders (MPD), which is believed to underlie growth factor hypersensitivity displayed by haematopoietic progenitors in these disorders. However, its frequency has been rarely determined in Taiwanese patients. METHODS: The frequency of JAK2-V617F mutation in patients with polycythaemia vera, essential thrombocythaemia and idiopathic myelofibrosis (IMF) was determined in the DNA from the peripheral blood leucocytes of 108 patients by genomic polymerase chain reaction and restriction enzyme-based assay. RESULTS: The JAK2-V617F mutation could be detected in 28 of 33 polycythaemia vera patients (85%), 29 of 49 essential thrombocythaemia patients (59%) and 2 of 6 IMF patients (33%), but was not detected in 11 patients with myelodysplastic syndrome or another 10 with other haematological diseases. The presence of the JAK2 mutation was associated with specific MPD disease subtypes (P = 0.007), longer disease duration (P = 0.005), splenomegaly (P = 0.019), a higher white blood cell count (P = 0.002) and a higher haemoglobin level (P = 0.036). However, the overall risk of thrombosis or bleeding was not affected by the presence of the JAK2 mutation (32 vs 17%; P = 0.22). CONCLUSION: The JAK2-V617F mutation can be frequently detected in the Taiwanese patients with MPD disorders and therefore should be incorporated into the initial evaluation of patients suspected of MPD.

Direct assessment of international consistency of standards for ground-level ozone: strategy and implementation toward metrological traceability network in Asia.

An international exercise to directly assess consistency of standards for ground-level ozone in East Asia was conducted as part of the East Asian Regional Experiment 2005 (EAREX 2005) in the framework of the Atmospheric Brown Clouds (ABC) project. Ten organizations collaboratively participated in the intercomparison. Four groups representing Japan, Korea, Hong Kong, and Taiwan made comparisons at the Gosan super observatory, Jeju Island, Korea, in March 2005, with ozone instruments calibrated to their national standards, and four Japanese groups made off-site comparisons with laboratory-level standards. All comparisons generally indicated good agreement with the standard reference photometer (SRP) 35, built by the National Institute of Standards and Technology (USA) and maintained by the National Institute for Environmental Studies (Japan). The assessment was expanded to measurement networks contributing to the World Meteorological Organization's Global Atmospheric Watch (WMO/GAW) program as part of off-site comparisons, and excellent agreement was achieved. These efforts contribute to propagating traceability of the national metrology standards among the atmospheric science community, to ensuring comparability of the existing ozone measurements, and to establishing an integrated network of air quality monitoring in Asia.

Increasing TB case detection through intensive referral of TB suspects by village doctors to county TB dispensaries.

OBJECTIVE: To explore new approaches to increase the detection of tuberculosis cases (TB). DESIGN: Thirty counties participated in the study. Patients with TB symptoms were surveyed and referred by trained village doctors to county dispensaries, designated township health centres or general hospitals for free sputum examination. TB patients and suspects notified by general hospitals were traced by TB staff if they defaulted during the transfer. RESULTS: A total of 12,091 new smear-positive TB cases were detected. The registration rate of new smear-positives increased from 36.2 per 100,000 population before the project to 49.9/100,000 after the project, and the case detection rate under the DOTS strategy reached 86%. Of 43,464 registered TB suspects, 15,363 (35.3%) were referred by village and hospital doctors. The referral rate increased significantly (P < 0.01). Of the 15,363 referred patients, 3870 were diagnosed as new smear-positive TB cases. Among three different microscopy centres, there was a statistically significant difference in the sputum examination rates of TB suspects and in the smear-positive rates among the suspects examined. The follow-up rate was 70.9%, but the follow-up success rate was only 33.1%. CONCLUSIONS: Intensive referral of patients with TB symptoms by village doctors to TB dispensaries is an effective way of increasing detection. At the same time, incentives are necessary for patients and village doctors.