The effect of white grub (Maladera Verticalis) larvae feeding on rhizosphere microbial characterization of aerobic rice (Oryza sativa L.) in Puer City, Yunnan Province, China.

BACKGROUND: Rhizosphere microorganisms are vital in plants' growth and development and these beneficial microbes are recruited to the root-zone soil when experiencing various environmental stresses. However, the effect of white grub (Maladera verticalis) larvae feeding on the structure and function of rhizosphere microbial communities of aerobic rice (Oryza sativa L.) is unclear. RESULTS: In this study, we compared physicochemical properties, enzyme activities, and microbial communities using 18 samples under healthy and M. verticalis larvae-feeding aerobic rice rhizosphere soils at the Yunnan of China. 16 S rRNA and ITS amplicons were sequenced using Illumina high throughput sequencing. M. verticalis larvae feeding on aerobic rice can influence rhizosphere soil physicochemical properties and enzyme activities, which also change rhizosphere microbial communities. The healthy and M. verticalis larvae-feeding aerobic rice rhizosphere soil microorganisms had distinct genus signatures, such as possible_genus_04 and Knoellia genera in healthy aerobic rice rhizosphere soils and norank_f__SC - I-84 and norank_f__Roseiflexaceae genera in M. verticalis larvae-feeding aerobic rice rhizosphere soils. The pathway of the metabolism of terpenoids and polyketides and carbohydrate metabolism in rhizosphere bacteria were significantly decreased after M. verticalis larvae feeding. Fungal parasite-wood saprotroph and fungal parasites were significantly decreased after M. verticalis larvae feeding, and plant pathogen-wood saprotroph and animal pathogen-undefined saprotroph were increased after larvae feeding. Additionally, the relative abundance of Bradyrhizobium and Talaromyces genera gradually increased with the elevation of the larvae density. Bacterial and fungal communities significantly correlated with soil physicochemical properties and enzyme activities, respectively. CONCLUSIONS: Based on the results we provide new insight for understanding the adaptation of aerobic rice to M. verticalis larvae feeding via regulating the rhizosphere environment, which would allow us to facilitate translation to more effective measures.

Genetic Diversity and Virulence Variation of Metarhizium rileyi from Infected Spodoptera frugiperda in Corn Fields.

Metarhizium rileyi is an entomopathogenic fungus that naturally infects the larvae of Spodoptera frugiperda, and has biocontrol potential. To explore more natural entomopathogenic fungi resources, a total of 31 strains were isolated from 13 prefectures in Yunnan Province. All the strains were identified using morphology and molecular biology. The genetic diversity of the 31 isolates of M. rileyi was analyzed using inter-simple sequence repeat (ISSR) techniques. Seven primers with good polymorphism were selected, and fifty-four distinct amplification sites were obtained by polymerase chain reaction amplification. Among them, 50 were polymorphic sites, and the percentage of polymorphic sites was 94.44%. The thirty-one strains were divided into eight subpopulations according to the regions. The Nei's gene diversity was 0.2945, and the Shannon information index was 0.4574, indicating that M. rileyi had rich genetic diversity. The average total genetic diversity of the subpopulations in the different regions was 0.2962, the gene diversity within the populations was 0.1931, the genetic differentiation coefficient was 0.3482 (>0.25), and the gene flow was 0.9360 (<1). The individual cluster analysis showed that there was no obvious correlation between the genetic diversity of the strains and their geographical origin, which also indicated that the virulence of the strains was not related to their phylogeny. Thus, the genetic distance of the different populations of M. rileyi in Yunnan Province was not related to the geographical distance. The virulence of those 32 strains against the 3rd-instar larvae of S. frugiperda were varied with the differences in geographical locations. On the 10th day of inoculation, seventeen strains had an insect mortality rate of 70.0%, and seven strains had an insect mortality rate of 100%. The half-lethal times of the M. rileyi SZCY201010, XSBN200920, and MDXZ200803 strains against the S. frugiperda larvae were less than 4 d. Thus, they have the potential to be developed into fungal insecticidal agents.

Mechanism of the Change in the Intestinal Microbiota of C-Strain Spodoptera frugiperda (Lepidoptera: Noctuidae) after an Interspecific Transference between Rice and Corn.

Spodoptera frugiperda (J.E.Smith) (Lepidoptera: Noctuidae) was first found in 2019 in Yunnan, China, and it was characterized as a corn strain; it was also found on rice strains there, and it damages rice in China, but little is known about the effect of host plant transfer on the intestinal microbiota and the activities of detoxification enzymes in the C-strain (corn strain) S. frugiperda. The intestinal microbiota and the protective enzyme activity of S. frugiperda that were transferred from rice plants were assessed, and the fourth generation of insects transferred from corn were studied; the gene types of S. frugiperda that were transferred from rice plants were tested using mitochondrial Tpi gene sequences. The results showed that the intestinal microbiota in the C-strain S. frugiperda were changed after the host transference, and the diversity and richness of the intestinal bacterial communities of the S. frugiperda feeding on rice were significantly reduced after the transfer of the host from corn. The predominant species of intestinal bacteria of the S. frugiperda on rice transferred from corn were Enterococcus and Enterobacter, with relative abundances of 28.7% and 66.68%; the predominant species of intestinal bacteria of the S. frugiperda that were transferred from rice and feeding on corn were Enterococcus (22.35%) and Erysipelatoclostridium (73.92%); and the predominant species of intestinal bacteria of S. frugiperda feeding on corn was Enterococcus, with a relative abundance of 61.26%. The CAT (catalase) activity of the S. frugiperda transferred from corn onto rice from corn was reduced, the POD (peroxidase) activity was significantly increased after the transfer from corn, and no significant variations were found for the SOD (superoxide dismutase), CarE (carboxylesterase), and GST (glutathione S-transferase) activities of S. frugiperda after the host plant transfer. The results showed that after feeding on rice, the activities of CAT and POD in the in S. frugiperda body changed in order to resist plant secondary metabolites from corn or rice, but there was no significant change in the detoxification enzymes in the body. In summary, switching the host plant between corn and rice induced variations in the intestinal microbiota in C-strain S. frugiperda owing to the strain difference between the C-strain and the R-strain (rice strain), and this was consistent with the results of the activities of detoxification enzymes. The results indicat that changes in intestinal microbiota and physiological enzymes may be important reasons for the adaptive capacity of C-strain S. frugiperda to rice.

Reconstruction of Gut Bacteria in Spodoptera frugiperda Infected by Beauveria bassiana Affects the Survival of Host Pest.

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a migratory agricultural pest that is devastating on a global scale. Beauveria bassiana is a filamentous entomopathogenic fungus that has a strong pathogenic effect on Lepidoptera pests but little is known about the microbial community in the host gut and the dominant populations in fungus-infected insects. B. bassiana AJS91881 was isolated and identified from the infected larvae of Spodoptera litura. The virulence of AJS91881 to the eggs, larvae, pupae and adults of S. frugiperda was measured. Moreover, the gut microbial community diversity of healthy and fungus-infected insects was analyzed. Our results showed that after treatment with B. bassiana AJS91881, the egg hatching rate, larval survival rate and adult lifespan of the insects were significantly reduced, and the pupae rigor rate was significantly increased compared to that of the control group. Additionally, the gut microbial community was reconstructed after B. bassiana infection. At the phylum and genus level, the relative abundance of the Proteobacteria and Serratia increased significantly in the B. bassiana treatment group. The KEGG function prediction results showed that fungal infection affected insect gut metabolism, environmental information processing, genetic information processing, organism systems and cellular processes. Fungal infection was closely related to the metabolism of various substances in the insect gut. Serratia marcescens was the bacterium with the highest relative abundance after infection by B. bassiana; intestinal bacteria S. marcescens inhibited the infection of insect fungi B. bassiana against the S. frugiperda. The presence of gut bacteria also significantly reduced the virulence of the fungi against the insects when compared to the group with the larvae fed antibiotics that were infected with fungal suspension (Germfree, GF) and healthy larvae that were infected with fungal suspension prepared with an antibiotic solution (+antibiotic). In conclusion, the reconstruction of the insect intestinal bacterial community is an indispensable link for understanding the pathogenicity of B. bassiana against S. frugiperda. Most importantly, in the later stage of fungal infection, the increased abundance of S. marcescens in the insect intestine inhibited the virulence of B. bassiana to some extent. The findings aid in understanding changes in the gut microbiota during the early stages of entomopathogenic fungal infection of insects and the involvement of insect gut microbes in host defense mediated by pathogenic fungal infection. This study is also conducive to understanding the interaction between entomopathogenic fungi, hosts and gut microbes, and provides a new idea for the joint use of entomopathogenic fungi and gut bacteria to control pests.

Virulence of Metarhizium rileyi Is Determined by Its Growth and Antioxidant Stress and the Protective and Detoxifying Enzymes of Spodoptera frugiperda.

Spodoptera frugiperda is one of the most destructive crop pests in the world. Metarhizium rileyi is an entomopathogenic fungus specific for noctuid pests and is a very promising prospect in biological control against S. frugiperda. Two M. rileyi strains (XSBN200920 and HNQLZ200714) isolated from infected S. frugiperda were used to evaluate the virulence and biocontrol potential to different stages and instars of S. frugiperda. The results showed that XSBN200920 was significantly more virulent than HNQLZ200714 to eggs, larvae, pupae, and adults of S. frugiperda. In the larvae infected with the two M. rileyi strains, the activity of three protective enzymes (including peroxidase (POD), superoxide dismutase (SOD), catalase (CAT)) and two detoxifying enzymes (including glutathione-S transferase (GST) and carboxylesterase (CarE)) increased firstly and then decreased. The expression levels of protective enzymes and detoxification enzymes in larvae treated with XSBN200920 were greater than with HNQLZ200714. Furthermore, antioxidant stress-related gene (MrSOD and MrCAT family genes) expression in the two strains was measured by RT-qPCR (real-time quantitative PCR). The expression of these genes was significantly higher in the XSBN200920 strain compared to HNQLZ200714. There were also significant differences in the sensitivity of the two strains to the growth of different carbon and nitrogen sources and oxidative stress agents. In addition, the activity expression of antioxidant enzymes on the third day of culturing in XSBN200920 was significantly higher than with HNQLZ200714. In summary, the high virulence of M. rileyi XSBN200920 was not only determined by the expression levels of protective and detoxifying enzymes of the host but also regulated by the growth of entomogenic fungi and the resistance to the oxidative stress against S. frugiperda at different stages and instars. This study provides a theoretical fundament for the systematic control of Spodoptera frugiperda using Metarhizium rileyi.

Temporal and Spatial Distribution Patterns of Spodoptera frugiperda in Mountain Maize Fields in China.

Spodoptera frugiperda (Lepidoptera: Noctuidae) is a major pest of maize worldwide. This pest colonized maize in Shizong, Qujing, Yunnan, China in 2019. To explore the temporal and spatial distribution of S. frugiperda in local fields, "W" type 5-point sampling was performed from 2020 to 2021. The spatial distribution was analyzed using the aggregation index, Iwao's regression, and Taylor's power law. The temporal distribution showed two peaks for both 2020 and 2021 when the density of eggs, larvae, and adults was high throughout the maize growth period. Additionally, 1st and 3rd instar larvae were higher in number during the maize seedling, jointing, and spinning stages. Fourth to 6th instar larvae were higher in number after the tasseling stage. Additionally, the spatial distribution results showed that 1st to 3rd instar larvae were aggregated, while 4th to 6th instar larvae were uniformly distributed in mountain maize fields. This study provides monitoring data for S. frugiperda and clarifies the temporal and spatial distribution characteristics for larvae in mountain maize fields. Further, it also provides guidance for investigation into population dynamics and the development of predictive models for integrated S. frugiperda management.

A multifunctional enzyme portfolio for α-chaconine and α-solanine degradation in the Phthorimaea operculella gut bacterium Glutamicibacter halophytocola S2 encoded in a trisaccharide utilization locus.

Steroidal glycoalkaloids (SGAs) are secondary metabolites commonly found in members of the family Solanaceae, including potatoes, and are toxic to pests and humans. The predominant SGAs in potato are α-chaconine and α-solanine. We previously reported that Glutamicibacter halophytocola S2, a gut bacterium of the pest Phthorimaea operculella (potato tuber moth), can degrade α-chaconine and α-solanine in potatoes, which can improve the fitness of P. operculella to feed on potatoes with a high content of toxic SGAs. Glutamicibacter halophytocola S2 harbored a gene cluster containing three deglycosylase genes-GE000599, GE000600, and GE000601-that were predicted encode α-rhamnosidase (RhaA), ß-glucosidase (GluA), and ß-galactosidase (GalA). However, there is limited information is available on the enzyme activities of the three enzymes expressed by this gene cluster and how they degrade the major toxic α-chaconine and α-solanine. In the current study, each enzyme of this gene cluster was produced by a prokaryotic expression approach and the activity of the recombinant enzymes for their target substrate and α-chaconine and α-solanine were evaluated by EPOCH microplate spectrophotometer and liquid chromatography mass spectrometry (LC-MS). The three enzymes had multifunctional activities, with RhaA and GluA could hydrolyze α-rhamnose, ß-glucose, and ß-galactose, while GalA can hydrolyze ß-glucose and ß-galactose. The degradation of α-chaconine and α-solanine was consistent with the results of the enzyme activity assays. The final product solanidine could be generated by adding RhaA or GluA alone. In conclusion, this study characterized the multifunctional activity and specific degradation pathway of these three enzymes in G. halophytocola S2. The three multifunctional enzymes have high glycosidic hydrolysis activity and clear gene sequence information, which help facilitates understanding the detoxification mechanism of insect gut microbes. The enzymes have a broad application potential and may be valuable in the removal of toxic SGAs from for potato food consumption.

The Acaricidal Potential of a New Agent GC16 for Tetranychus pueraricola (Acari: Tetranychidae) Based on Developmental Performance and Physiological Enzyme Activity.

The spider mite, Tetranychus pueraricola (Ehara & Gotoh; Acari: Tetranychidae), is a serious pest in agriculture and horticulture. Application of chemical pesticides is the main mode of this pest control. Due to pesticide residues and resistance-induced resurgence of pests, there is a need to discover alternatives for spider mite management. GC16 comprises a mixture of calcium chloride (CaCl2, 45%) and lecithin (55%), which was recently found to have acaricidal properties. We evaluated the sublethal effects of GC16 on T. pueraricola using life table and enzyme [catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), carboxylesterase (CarE), glutathione S-transferases (GST), and Ca2+-ATPase (Ca2+-ATP)] activity assays. The results showed that fecundity of T. pueraricola increased at LC30 but decreased at LC50 of GC16. The intrinsic rate of increase (r) of T. pueraricola decreased under the LC30 and LC50 of GC16. GC16 concentration and exposure time significantly influenced the activities of CAT, POD, CarE, GST, and Ca2+-ATP in adult mites. Twelve hours later after the treatment, GST and Ca2+-ATP activities were significantly inhibited by LC30 but enhanced by LC50. Moreover, the demographic parameter r and enzyme activities were negatively correlated. In sum, sublethal amounts of GC16 had an adverse effect on mites, and there was a trade-off between developmental performance and physiological enzyme activity of mites under GC16 stress, and GC16 showed an acaricidal potential for T. pueraricola. This work provides guidance for the application of GC16 to control T. pueraricola.

The Insecticidal Efficacy and Physiological Action Mechanism of a Novel Agent GC16 against Tetranychus pueraricola (Acari: Tetranychidae).

Chemical control plays a crucial role in pest management but has to face challenges due to insect resistance. It is important to discover alternatives to traditional pesticides. The spider mite Tetranychus pueraricola (Ehara & Gotoh) (Acari: Tetranychidae) is a major agricultural pest that causes severe damage to many crops. GC16 is a new agent that consists of a mixture of Calcium chloride (CaCl2) and lecithin. To explore the acaricidal effects and mode of action of GC16 against T. pueraricola, bioassays, cryogenic scanning electron microscopy (cryo-SEM) and transmission electron microscopy (TEM) were performed. GC16 had lethal effects on the eggs, larvae, nymphs, and adults of T. pueraricola, caused the mites to dehydrate and inactivate, and inhibited the development of eggs. GC16 displayed contact toxicity rather than stomach toxicity through the synergistic effects of CaCl2 with lecithin. Cryo-SEM analysis revealed that GC16 damaged T. pueraricola by disordering the array of the cuticle layer crest. Mitochondrial abnormalities were detected by TEM in mites treated by GC16. Overall, GC16 had the controlling efficacy on T. pueraricola by cuticle penetration and mitochondria dysfunction and had no effects on Picromerus lewisi and Harmonia axyridis, indicating that GC16 is likely a new eco-friendly acaricide.

Glutamicibacter halophytocola-mediated host fitness of potato tuber moth on Solanaceae crops.

BACKGROUND: The potato tuber moth (PTM), Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), is a destructive pest of Solanaceae crops worldwide. α-solanine and α-chaconine are toxic steroidal glycoalkaloids (SGAs) in Solanaceae crops and are most abundant in potatoes (Solanum tuberosum L.), accounting for more than 95% of the total SGAs. PTM grows on potatoes with a higher concentration of SGAs. Gut bacteria play an important role in the physiology and behavior of insects. To understand the role of gut bacteria of PTM in host adaptability, we isolated and identified major SGA (α-chaconine and α-solanine)-degrading gut bacteria in the gut of PTM by a selective medium and analyzed their degradability and degradation mechanism. RESULTS: The gut Glutamicibacter halophytocola S2 of PTM with high degradation capacity to α-solanine and α-chaconine were detected by liquid chromatography mass spectrometry (LC-MS) and identified by morphological and 16S rRNA gene sequence analysis. A gene cluster involving α-rhamnosidases, ß-glucosidases, and ß-galactosidases was identified by whole-genome sequencing of G. halophytocola S2. These genes had higher expression on the α-solanine medium. PTM inoculated with the isolated G. halophytocola S2 obtained higher fitness than antibiotic-treated PTM. CONCLUSION: The G. halophytocola S2 in the gut of PTM could degrade the major toxic α-solanine and α-chaconine in potatoes. This enhances the fitness of PTM feeding on potatoes with high SGA contents. The results provide a theoretical foundation for the integrated pest management of PTM and provide an effective strain for the treatment of α-solanine and α-chaconine in potato food. © 2022 Society of Chemical Industry.

Midgut microbiota diversity of potato tuber moth associated with potato tissue consumed.

BACKGROUND: The potato tuber moth (PTM), Phthorimaea operculella (Zeller), is a worldwide pest that feeds on both the leaves and tubers of potato plants. PTM larvae can digest leaves, or tubers, resulting in serious damage to potato plants in the field and potato tubers in storage. To understand how midgut bacterial diversity is influenced by the consumption of these two tissue types, the symbiotic bacteria in the potato-feeding PTM midgut and the endophytic bacteria of potato tissues were analyzed. RESULTS: At the genus level, the bacterial community composition in the PTM midgut was influenced by the tissues consumed, owing to their different nutrient contents. Escherichia_Shigella and Enterobacter were the most dominant genera in the midgut of leaf-feeding and tuber-feeding PTMs, respectively. Interestingly, even though only present in low abundance in leaves and tubers, Escherichia_Shigella were dominantly distributed only in the midgut of leaf-feeding PTMs, indicating that specific accumulation of these genera have occurred by feeding on leaves. Moreover, Enterobacter, the most dominant genus in the midgut of tuber-feeding PTMs, was undetectable in all potato tissues, indicating it is gut-specific origin and tuber feeding-specific accumulation. Both Escherichia_Shigella and Enterobacter abundances were positively correlated with the dominant contents of potato leaves and tubers, respectively. CONCLUSIONS: Enrichment of specific PTM midgut bacterial communities was related to different nutrient levels in different tissues consumed by the insect, which in turn influenced host utilization. We provide evidence that a portion of the intestinal microbes of PTMs may be derived from potato endophytic bacteria and improve the understanding of the relationship between potato endophytic bacteria and the gut microbiota of PTMs, which may offer support for integrated management of this worldwide pest.

The Time-Concentration-Mortality Responses of Western Flower Thrips, Frankliniella occidentalis, to the Synergistic Interaction of Entomopathogenic Fungus Metarhizium flavoviride, Insecticides, and Diatomaceous Earth.

Western flower thrips (WFT), Frankliniella occidentalis (Pergande), is a highly invasive pest which is harmful to many cash crops globally and resistant to various insecticides. Entomopathogenic fungi (EPF), as biological control agents, have demonstrated a good control effect on WFT. The aim of this study was to evaluate the synergistic and pathogenicity efficacy of the fungal strain Metarhizium flavoviride WSWL51721 when distributed with diatomaceous earth (DE) and the active ingredient imidacloprid using four bioassay methods against adult and second instar larvae of WFT. The data of the four bioassays have been fitted to the time-concentration-mortality (TCM) model. The corrected mortality ranges of WFT adults were 75-100%, 82.69-100%, 78.85-100%, and 92.31-100%, and the corrected mortality ranges of WFT second instar larvae were 72.22-100%, 85.19-100%, 77.77-100%, and 100% in the four bioassays at concentrations of 1.2 × 106 to 1.2 × 108 conidia/mL, respectively. At 1.2 × 108 conidia/mL, assays 2 (M. flavoviride with DE), 3 (M. flavoviride with imidacloprid), and 4 (M. flavoviride with DE and imidacloprid) had the shortest median lethal time (LT50), compared with that of assay 1 (M. flavoviride alone) for adults at 2.26 d, 2.06 d, and 1.53 d, and second instar larvae at 2.45 d, 1.70 d, and 1.41 d, respectively. The median lethal concentration (LC50) in the four bioassays decreased within 3-10 days of inoculation. On the third day, it was found that the lowest median lethal concentrations in assays 2, 3, and 4 were 1.58 × 107, 1.13 × 107, and 3.39 × 106 conidia/mL, respectively, which were significantly different from that in assay 1 for the adults. For the second instar larvae, assays 2, 3, and 4 also had the lowest lethal concentrations and were significantly different from those of assay 1. There were significant differences in sporulation between adults and second instar larvae under the four bioassays. Our results indicate that assays 2 (M. flavoviride with DE), 3 (M. flavoviride with imidacloprid), and 4 (M. flavoviride with DE and imidacloprid) demonstrate synergistic effects on the control of both adult and second instar larvae of WFT under laboratory conditions.

[Efficacy of NO regimen and NP regimen on advanced non-small cell lung cancer: a prospective randomized trial].

BACKGROUND & OBJECTIVE: Oxaliplatin (LOHP) is an effective drug in treatment of non-small cell lung cancer (NSCLC) with mild toxicities to gastrointestinal tract, kidney, and bone marrow. Cisplatin (DDP) plus vinorelbine (NVB) constitute the first-line regimen (NP regimen) for NSCLC. This study was to compare the short-term response, long-term outcome, and adverse events between advanced NSCLC patients received NO regimen (LOHP plus NVB) and NP regimen. METHODS: A total of 90 patients with advanced NSCLC were randomized into NO group (58 patients, 25 mg/m(2) of NVB, day 1 and day 8; 130 mg/m(2) of LOHP, day 1) and NP group (32 patients, 25 mg/m(2) of NVB, day 1 and day 8; 50 mg/m(2) of DDP, day 2 and day 3). The short-term response, long-term outcome, adverse events, and survival status of the 2 groups were observed. RESULTS: The response rates were 33.33% in NO group, and 35.48% in NP group, but no significant difference was detected between the 2 groups (P > 0.05). The clinical benefit response rate was significantly higher in NO group than in NP group (80.70% vs. 64.52%, P < 0.05). The median time to progression (TTP) was 17 weeks in NO group, and 15 weeks in NP group; the median time of remission was 21 weeks in NO group, and 19 weeks in NP group; the median survival time was 39 weeks in NO group, and 37 weeks in NP group; the 1-year survival rate was 37.93% in NO group, and 31.25% in NP group. No significant differences were detected between the 2 groups. The incidence rates of phlebitis and grade I-II peripheral neuritis were significantly higher in NO group than in NP group (77.59% vs. 50.00%, P<0.01; 43.10% vs. 15.63%, P<0.01). The incidence rate of grade III-IV nausea/vomiting was significantly higher in NP group than in NO group (31.25% vs. 3.45%, P<0.05). CONCLUSIONS: The efficacy of NO regimen on advanced NSCLC is similar to that of NP regimen, but the clinical benefit response rate is higher in NO group than in NP group. In short, NO regimen may be recommended as the first-line chemotherapy regimen for advanced NSCLC.

[Prospective randomized study of HMVP, MVP, and HVP regimens in treatment of advanced non-small cell lung cancer].

BACKGROUND & OBJECTIVE: Non-small cell lung cancer (NSCLC) is hyposensitive to the normal first and second-line chemotherapy regimens. Camptothecin derivative is becoming a hot point in the treatment of advanced NSCLC. The objective of this article was to evaluate the response, toxicity, and survival time of HMVP, MVP, and HVP regimens (detail in below) in the treatment of advanced NSCLC. METHODS: A total of 134 cases with advanced NSCLC was randomized into three groups: HMVP group [46 patients, hydroxycamptothecin (HCPT) 12 mg/m(2) from d1 to d5, mitomycin C (MMC) 6 mg/m(2) d1, vindesine (VDS) 2.5-3 mg/m(2) d1 and d8, cisplatin (DDP) 50 mg/m(2) d2 and d3], MVP group (44 patients, MMC, VDS and DDP were the same as HMVP group) and HVP group (44 patients, HCPT, VDS, DDP were the same as HMVP group). RESULTS: The response rates were 39.54% (17/43), 35.57% (15/42), and 26.19% (11/42) in HMVP, MVP, and HVP groups, respectively; no significant difference was detected among the three groups (P >0.05). No significant difference was detected in the median time of remission, median survival time, and 1-, 2-year survival rates among the three groups. Moreover, no significant difference was detected in grade III-IV leukopenia, grade III-IV thrombocytopenia, grade III-IV nausea and vomiting and grade III-IV constipation among the three groups. CONCLUSION: The response rate of MVP regimen is slightly lower than that of HMVP regimen, but HMVP regimen do not show obvious superiority. It may increase toxicities such as leukopenia, nausea/vomiting, and constipation. The response rate of HVP regimen is slightly lower than that of MVP regimen.