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BackgroundLenvatinib monotherapy and combination therapy with immune checkpoint inhibitors (ICI) were widely applied for unresectable hepatocellular carcinoma (uHCC). However, many patients failed to benefit from the treatments. A prognostic model was needed to predict the treatment outcomes and guide clinical decisions.Methods304 patients receiving lenvatinib monotherapy or lenvatinib plus ICI for uHCC were retrospectively included. The risk factors derived from the multivariate analysis were used to construct the predictive model. The C-index and area under the receiver-operating characteristic curve (AUC) were calculated to assess the predictive efficiency.ResultsMultivariate analysis revealed that protein induced by vitamin K absence or antagonist-II (PIVKA-II) (HR, 2.05; P=0.001) and metastasis (HR, 2.07; P600 mAU/mL) and PIMET-high group (with metastasis and PIVKA-II>600 mAU/mL). The C-index of PIMET score for the survival prediction was 0.63 and 0.67 in the training and validation cohort, respectively. In the training cohort, the AUC of 12-, 18-, and 24-month OS was 0.661, 0.682, and 0.744, respectively. The prognostic performances of the model were subsequently validated. The AUC of 12-, 18-, and 24-month OS was 0.724, 0.726, and 0.762 in the validation cohort. Subgroup analyses showed consistent predictive value for patients receiving lenvatinib monotherapy and patients receiving lenvatinib plus ICI. The PIMET score could also distinguish patients with different treatment responses. Notably, the combination of lenvatinib and ICI conferred survival benefits to patients with PIMET-int or PIMET-high, instead of patients with PIMET-low.ConclusionThe PIMET score comprising metastasis and PIVKA-II could serve as a helpful prognostic model for uHCC receiving lenvatinib monotherapy or lenvatinib plus ICI. The PIMET score could guide the treatment decision and facilitate precision medicine for uHCC patients.

BackgroundImmune checkpoint inhibitor (ICI)-based combination therapy has opened a new avenue for the treatment of multiple malignancies including hepatocellular carcinoma (HCC). However, considering the unsatisfactory efficacy, biomarkers are urgently needed to identify the patients most likely to benefit from ICI-based combination therapy.MethodsA total of 194 patients undergoing ICI-based combination therapy for unresectable HCC were retrospectively enrolled and divided into a training cohort (n = 129) and a validation cohort (n = 65) randomly. A novel circulating immune index (CII) defined as the ratio of white blood cell count (×109/L) to lymphocyte proportion (%) was constructed and its prognostic value was determined and validated.ResultsPatients with CII ≤ 43.1 reported prolonged overall survival (OS) compared to those with CII > 43.1 (median OS: 24.7 vs 15.1 months; 6-, 12-, 18-month OS: 94.2%, 76.7%, 66.1% vs 86.4%, 68.2%, 22.8%, P = 0.019), and CII was identified as an independent prognostic factor for OS (hazard ratio, 2.24; 95% confidence interval, 1.17-4.31; P = 0.015). These results were subsequently verified in the validation cohort. Additionally, patients with low CII levels had improved best radiological tumor response (complete response, partial response, stable disease, progressive disease: 3%, 36%, 50%, 11% vs 0%, 27%, 46%, 27%; P = 0.037) and disease control rate (89% vs 73%; P = 0.031) in the pooled cohort and better pathologic response (pathologic complete response, major pathologic response, partial pathologic response, no pathologic response: 20%, 44%, 28%, 8% vs 0%, 0%, 40%, 60%; P = 0.005) in the neoadjuvant cohort. Detection of lymphocyte subsets revealed that an elevated proportion of CD4+ T cells was related to better OS, while the proportion of CD8+ T cells was not.ConclusionsWe constructed a novel circulating immune biomarker that was capable of predicting OS and therapeutic efficacy for HCC patients undergoing ICI and lenvatinib combination therapy.

Hepatocellular carcinoma (HCC) accounts for the predominant form of liver malignancy and presents a leading cause of cancer-related death globally. Sorafenib (SOR), a first-line targeted drug for advanced HCC treatment, has a battery of untoward side effects. Photothermal therapy (PTT) has been utilized as an effective adjuvant in synergy with other approaches. However, little is known about the tumoricidal efficacy of combining SOR with PTT for HCC. Herein, a novel versatile nanoparticle, Cu

BackgroundTumor recurrence after hepatectomy is high for hepatocellular carcinoma (HCC), and minimal residual disease (MRD) could be the underlying mechanism. A predictive model for recurrence and presence of MRD is needed.MethodsCommon inflammation-immune factors were reviewed and selected to construct novel models. The model consisting of preoperative aspartate aminotransferase, C-reactive protein, and lymphocyte count, named ACLR, was selected and evaluated for clinical significance.ResultsAmong the nine novel inflammation-immune models, ACLR showed the highest accuracy for overall survival (OS) and time to recurrence (TTR). At the optimal cutoff value of 80, patients with high ACLR (> 80) had larger tumor size, higher Edmondson’s grade, more vascular invasion, advanced tumor stage, and poorer survival than those with low ACLR (≤ 80) in the training cohort (5-year OS: 43.3% vs. 80.1%, P < 0.0001; 5-year TTR: 74.9% vs. 45.3%, P < 0.0001). Multivariate Cox analysis identified ACLR as an independent risk factor for OS [hazard ratio (HR) = 2.22, P < 0.001] and TTR (HR = 2.36, P < 0.001). Such clinical significance and prognostic value were verified in validation cohort. ACLR outperformed extant models, showing the highest area under receiver operating characteristics curve for 1-, 3-, and 5-year OS (0.737, 0.719, and 0.708) and 1-, 3-, and 5-year TTR (0.696, 0.650, and 0.629). High ACLR correlated with early recurrence (P < 0.001) and extremely early recurrence (P < 0.001). In patients with high ACLR, wide resection margin might confer survival benefit by decreasing recurrence (median TTR, 25.5 vs. 11.4 months; P = 0.037).ConclusionsThe novel inflammation-immune model, ACLR, could effectively predict prognosis, and the presence of MRD before hepatectomy and might guide the decision on resection margin for patients with HCC.

Lenvatinib is one of the first-line treatments for unresectable hepatocellular carcinoma (HCC). However, data are lacking on lenvatinib in the postoperative setting.This retrospective analysis enrolled 242 patients with HCC who underwent liver transplantation (LTx). Eligible patients were divided into 2 groups according to their use of adjuvant lenvatinib following LTx (lenvatinib, n=42; control, n=200). The primary outcome measures were overall survival (OS), time to recurrence (TTR), and safety. Kaplan-Meier analysis was applied to calculate the OS, while a competing risk model was used to estimate the cumulative incidence of recurrence.The lenvatinib group showed more advanced tumors and a higher proportion of HCC beyond the Milan criteria (P0.001) than the control group. There were no significant differences in both the OS and TTR between the 2 groups. After focusing on the patients with HCC beyond the Milan criteria, baseline characteristics were similar in the lenvatinib group (n=38) and the control group (n=102). Competing risk analysis showed lenvatinib significantly prolonged TTR after LTx versus the control group [sub-hazard ratio (sHR), 0.40; 95% confidence interval (CI): 0.17 to 0.93; P=0.031]. In the multivariate competing risk model, adjuvant lenvatinib was an independent protective factor for tumor recurrence after LTx in patients with HCC beyond the Milan criteria (sHR, 0.33; 95% CI: 0.13 to 0.83; P=0.018). The rate of early recurrence within t2 years after LTx was also significantly decreased in the lenvatinib group (15.8%Postoperative lenvatinib administration may provide clinical benefits and is well tolerated in patients with HCC beyond the Milan criteria who undergo LTx.

Hepatocellular carcinoma (HCC) accounts for the predominant form of liver malignancy and presents a leading cause of cancer-related death globally. Sorafenib (SOR), a first-line targeted drug for advanced HCC treatment, has a battery of untoward side effects. Photothermal therapy (PTT) has been utilized as an effective adjuvant in synergy with other approaches. However, little is known about the tumoricidal efficacy of combining SOR with PTT for HCC. Herein, a novel versatile nanoparticle, Cu2−xSe@SOR@PEG (CSP), that is based on a photothermal Cu2−xSe core and SOR for simultaneously reinforcing PTT and reducing the adverse effects of SOR was constructed. The synthesized CSP exhibited a remarkably enhanced therapeutic effect upon 808 nm laser irradiation via dampening HCC cell propagation and metastasis and propelling cell apoptosis. The intravenous administration of CSP substantially suppressed tumor growth in a xenograft tumor mouse model. It was noted that the CSP manifested low toxicity and excellent biocompatibility. Together, this work indicates a promising and versatile tool that is based on synergistic PTT and molecular-targeted therapy for HCC management.

Pancreatic cancer is an increasingly common cause of cancer mortality with a tight correspondence between disease mortality and incidence. Furthermore, it is usually diagnosed at an advanced stage with a very dismal prognosis. Due to the high heterogeneity, metabolic reprogramming, and dense stromal environment associated with pancreatic cancer, patients benefit little from current conventional therapy. Recent insight into the biology and genetics of pancreatic cancer has supported its molecular classification, thus expanding clinical therapeutic options. In this review, we summarize how the biological features of pancreatic cancer and its metabolic reprogramming as well as the tumor microenvironment regulate its development and progression. We further discuss potential biomarkers for pancreatic cancer diagnosis, prediction, and surveillance based on novel liquid biopsies. We also outline recent advances in defining pancreatic cancer subtypes and subtype-specific therapeutic responses and current preclinical therapeutic models. Finally, we discuss prospects and challenges in the clinical development of pancreatic cancer therapeutics.

Nd2Fe14B rare earth and Sm2Co17 type permanent magnets have been widely used in the third generation of synchronous radiation light source and free electron laser facility in undulators and other components of particle accelerators. In addition, the permanent magnets are used in the radiation treatment system for cancer as a beam line component. Compared with Sm2Co17 type permanent magnet, Nd2Fe14B rare earth permanent magnet has the characteristics of large magnet energy product, rich starting materials and low price. Although its Curie point and coercive force are lower than those of Sm2Co17 type of permanent magnet, Nd2Fe14B rare earth permanent magnet is still widely used. As an important part of the accelerator, the magnetic loss phenomenon appears when permanent magnet is used in long-term irradiation environments, which affects the stability and quality of the beam. Therefore, it is important to investigate the magnet demagnetization induced by photon irradiation. Recently, there have appeared many researches of the phenomena of demagnetization for the permanent magnets under the irradiation of various kinds of particles. By using different research methods and experimental conditions, single particle irradiation is performed and then the effect of irradiation on magnetic loss is investigated by comparing the macro magnetic properties (such as magnetic flux loss rate, saturation magnetization, etc.). However, there are not any available reports on the microstructure investigations of permanent magnets after irradiation. Microstructure affects macroscopic magnetic properties. In order to discuss the microscopic demagnetization mechanism, the transmission electron microscope is used to characterize and analyze the microstructure evolutions of Sm2Co17 type permanent magnet and Nd2Fe14B rare earth permanent magnet before and after proton irradiation. The evolution of the number density of nanocrystal and its size distribution induced by proton irradiation are calculated. Moreover, the effect of microstructure evolution on macroscopic magnetic loss is discussed. The results indicate that the microstructure of permanent magnet transforms from single crystal structure to polycrystalline structure with the increase of the proton irradiation damage level. Nanocrystal and the matrix of permanent magnet have the same crystal structure. With the irradiation damage level increasing, the nanocrystal density of Nd2Fe14B first increases and then decreases, while the particle size distribution first increases and then keeps constant; the number density of nanocrystal of Sm2Co17 type permanent magnet gradually decreases, while particle size gradually increases, and comparing with Sm2Co17 type permanent magnet, the crystal structure of Nd2Fe14B permanent magnet shows an obvious tendency to be amorphous in 2 dpa irradiation damage level.

CaAl12O19:xMn(4+) (x=0.0~0.00184 mol) red phosphors were prepared by high-temperature solid state method. Different influencing factors, i. e. the doping amount of Mn(4+), and sintering temperature, were investigated by the orthogonal experiment design. The optimum conditions for preparing CaAl12O19:xMn(4+) phosphors are: sintering temperature 1500 °C, and x=0.0147 mol. It emits strong deep red light with emission maximal peak at 656 nm, and two shoulder peaks (643 and 666 nm), the corresponding excitation peak is located at 470 nm. The 656 nm emission peak is ascribed to the (2)E--(4)A2 transition, and the shoulder peaks (643 and 666 nm) are attributed to phonon sideband transitions. The 470 nm excitation peak is ascribed to the ligand-field (4)A2--(4)T2 transition (d--d band). Owing to the fact that the excitation peak (470 nm) is near the emission (460 nm) of commercial LED chips, it is quite suitable for red light candidate for LED light resources.

Insertion devices are crucial parts of the third generation of synchrotron radiation facility and free electron laser devices. The use of insertion device can improve the brightness and coherence of synchrotron radiation light. Undulator, one kind of insertion device, is largely installed in the storage ring of Shanghai synchrotron radiation facility. The main part of undulator is the device of magnetic source which consists of periodically arranged permanent magnets with the same magnetic field strength. In order to keep the normal electronic trajectory, a stable magnetic intensity in undulator is required. The Sm2Co17 type permanent magnets with high intrinsic coercive force and good radiation stability are largely installed in the facility. However, the losses for magnetic properties of Sm2Co17 type permanent magnets can be induced by longperiod irradiation in undulator through beam loss or mis-steering. The reduction of magnetic field could affect the electron energy, direction and the movement trajectory and so on, which seriously affects the amount of synchrotron radiation light. Microstructure of Sm2Co17 type permanent magnet affects the macro magnetic properties and there is not any available report on the microstructure investigation of Sm2Co17 type permanent magnet after being irradiated. Therefore, in this work, the effect of irradiation on the microstructure evolution is investigated. The radiation fields of Sm2Co17 type permanent magnets and the main particles (neutron) that result in losing magnetic properties are first analyzed and confirmed by Monte Carlo code FLUKA calculation. Then, Sm2Co17 type permanent magnet samples are irradiated by Ar ions at different fluences to simulate neutron irradiation damage. Meanwhile, the microstructure evolutions of irradiated samples are characterized by transmission electron microscopy. Moreover, high resolution transmission electron microscopic images are taken at the peak of radiation damage field to further investigate the radiation damage. In the respect of macro magnetic properties, hysteresis loops are measured by vibrating sample magnetometer in order to study the change of saturated magnetization. The results indicate that the decrease of saturated magnetization value is related to the change of microstructure, which proves the speculation of previous investigations. The evolution of 2:17 phase transformed from single crystals into amorphous structure is a possible microscopic mechanism for irreversible loss for saturated magnetization of Sm2Co17.

Ethnopharmacological relevance Caesalpinia sappan is a medicinal plant native to China popularly used to treat chronic pelvic inflammation, dysmenorrhea and hysteromyoma. Its main bioactive component is brazilin which had presented antibacterial, anti-inflammatory and anti-platelet aggregation activities. To establish a sensitive, selective, reproducible, and accurate high performance liquid chromatographic (HPLC) method for the quantitative determination of brazilin in plasma, and study the pharmacokinetics of brazilin in rats after intravenous administration of brazilin. Materials and methods Rats received intravenous injection of 25, 50 and 100 mg/kg of brazilin. Concentrations of brazilin in plasma were determined by HPLC method at different time points and all pharmacokinetic parameters were estimated by non-compartmental analysis with WinNonLin 6.2 software. Results After single intravenous doses of 25, 50 and 100 mg/kg brazilin in rats, the main PK parameters were as follows: Cmax were 18.1±4.1, 46.7±8.7 and 82.2±9.6 µg/mL; AUC0–24 were 20.4±4.3, 48.7±6.8 and 90.4±10.3 µg h/mL; and t1/2 were 5.4±1.5, 5.8±0.9 and 6.2±1.2 h, respectively. Conclusion It showed that the brazilin was eliminated moderately in rat by intravenous injection route with t1/2 of 6 h and showed a dose-dependence profile of Cmax and AUC0–24 at the doses of 25~100 mg/kg of brazilin for injection in rats.

A convenient and rapid HPLC method was developed for the determination of clinofibrate in human plasma using simple protein precipitation with the mixture of acetonitrile and 1M hydrochloric acid (95:5, v/v) followed by separation using an Inspire C(18) column with isocratic elution. The detection wavelength was 232nm and the flow rate was 1.0ml/min. The mobile phase consisted of acetonitrile and water containing 0.4% ortho-phosphoric acid (73:27, v/v). Linear calibration curve was obtained over the concentrations ranging from 0.5μg/ml to 32μg/ml (r(2)=0.999) with LLOQ of 0.5μg/ml. The RSD in both the intra-run and inter-run precision study was less than 5.4% and the extraction recoveries were above 90.7%. The HPLC method is reproducible and suitable for the quantification of clinofibrate in plasma. This method was successfully applied to the pharmacokinetic studies of clinofibrate in healthy volunteers. The elimination half-lives (t(1/2)) were (20.47±3.44), (18.19±2.62) and (21.51±4.78)h after single oral administration of 200, 400 and 600mg clinofibrate, respectively. The results of WinNonlin software showed that the area under the plasma concentration versus time curve from time 0 to 72h (AUC(0-72)) and peak plasma concentration (C(max)) were linearly related to dose (P>0.05).

A high-performance liquid chromatographic method was developed for the determination of a new derivative of docetaxel, felotaxel, in rat plasma and human plasma and urine. The separation of felotaxel was performed on a Dikma C18 column with 0.2% formic acid and acetonitrile (50:50) as the mobile phase. The flow rate was 0.8 mL/min and the column effluent was monitored by an ultraviolet detector set at 275 nm. The method was validated and found to be linear in the range of 5-1,000 ng/mL. The lower limit of quantification was 5 ng/mL based on 100 µL of plasma. The variations for intra-day and inter-day precision were less than 6.9%, and the accuracy values were between 87.3 and 107.4%. The extraction recoveries were more than 80.5%. These data confirm that the developed method has satisfactory sensitivity, accuracy and precision for the quantification of felotaxel in rat plasma and in human plasma and urine. The method was successfully applied to a pharmacokinetics study of felotaxel after intravenous doses of 5 mg/kg in rats.

To investigate the effect of zinc sulfate on pharmacokinetics of cephalexin when administered concurrently or at strategically spaced dosing times designed to avoid the potential interaction in healthy volunteers.In this study, all subjects (n= 12) were randomized to receive the following four treatments, separated by a wash-out period of 7 days: cephalexin 500mg alone, concomitantly with zinc 250mg, 3h after zinc 250mg or 3h before zinc 250mg.All subjects completed the study safely. Zinc supplements administered concurrently with cephalexin significantly decreased the peak serum concentration (C(max) ), area under the plasma concentration-time curve from zero to infinity (AUC(0-∞) ) and the time for which the plasma concentration of the drug remained above the minimal inhibitory concentration of the pathogenic organism (TMIC) of cephalexin [mean percentage decrease (95% confidence intervals) of 31.05% (22.09-40.01%), 27.40% (18.33-36.47%) and 22.33% (12.51-32.16%), respectively; P0.05] compared with administration of cephalexin alone. Also, administration of zinc 3h before cephalexin decreased the C(max) , AUC(0-∞) and TMIC of the drug compared with administration of cephalexin alone [mean percentage decrease (95% confidence intervals) of 11.48% (3.40-19.55%), 18.12% (9.63-26.60%) and 23.75% (14.30-33.20%), respectively; P0.05]. In contrast, the pharmacokinetics of cephalexin was not notably altered by administration of zinc 3h after cephalexin dosing (P0.05).The significant interaction between zinc and cephalexin might affect the clinical outcome of cephalexin therapy. The dosing recommendation is that zinc sulfate can be safely administered 3h after a cephalexin dose.

Caenis pycnacantha sp. nov. Mature larvae (in alcohol): Body length 3.6���5.4 mm, caudal filaments 3.7���4.9 mm. Body generally pale to slightly pale brown or brown. Vertex dark brown with pale ecdysis suture. Ocelli pale, compound eyes dark black (Figs. 1, 12���13). Mouthparts: labrum slightly concave medially, and with dense short hair and spines on anterior margin and dorsal surface (Fig. 2). Mandibles: with long marginal setae on surface and seven to eight spines on inner margin. Left mandible (Fig. 3): outer incisor with three denticles while inner incisor with two; prostheca clavate, with long fine setae and two sharp bristles at apex. Right mandible (Fig. 4): both of the incisors with two denticles, the apex of prostheca represented by a tuft of branched setae. Maxillae (Fig. 5) with 3 obvious bristles at apex and scattered long, fine setae and spines along inner and outer margin. Maxillary palpi segment 2 is the shortest in the three segments (length ratio of them: 1: 0.7: 1.1); segment 1 and 3 with sparse long hair and short spines along both inner and outer margins while segment 2 with three long setae on outer margin subapically only. Segment 1 and 2 of labial palpi subequal in length, segment 3 shorter than any of them; segment 2 and 3 decorated with dense long setae. Glossae and paraglossae with dense tiny setae at apex (Fig. 6). Hypopharynx shown in figure 7, superlinguae with a row of long hair along free margin (Fig. 7). Thorax: Pronotum expanded outwards slightly, with three irregular pale spots dorsally, a narrow and shallow subanterior ridge located near anterior margin; anterolateral angles of pronotum expanded forwards slightly, lateral margin with sparse hairs. Mesonotum with small expanded anterolateral lobes, a pair of coneshaped pale dots near anterior margin; medial line prominent, forming a longitudinal ridge sometimes. Legs pale brown, with dense setae and spines; femora with blackish brown band apically; forefemur with a row of transverse spine-like setae near apex (Figs. 1, 12); Length ratio of femora: tibiae: tarsi of foreleg = 1.5: 1.0: 1.0; claw single, with 6 denticles at base. Tibiae and tarsi pale brown. Mid- and hindlegs with similar color pattern but without transverse setae on femora; mid claw with 8���10 denticles at base, hind claw with many denticles from base to subapex, basal 3 ones slightly larger. Abdomen: terga 1, 2, 10 darker than others; terga 7���9 with three longitudinal pale stripes, one median, two near middle line (Figs. 1, 12); terga 2���9 with sharp posterolateral projections; a row of strong and acute denticle on posterior margin of each sternum. Apex of sternum 9 with blunt margin. Operculate gill 2 subquadrate; dorsal surface pale-brown, color pattern similar to abdominal terga; Y-shape ranges distinct, with dense setae along free margins and few longer setae on lateral edge; ventral surface of gills 2 with tiny palmate outgrowths near outer and posterior margins (Figs. 14���15). Caudal filaments pale, with sparse setae between segments. Male imago (in alcohol): Body length 2.8 mm, caudal filaments 8.0 mm. Head and thorax dark brown; abdomen pale, with diffused brown marks (Fig. 16). Antennae pale, pedicel 1.5X as long as scape, flagellum expanded at base and tapered to apex (Fig. 8). Ocelli pale, compound eyes dark brown. Forefemur washed with brownish black markings; mid and hind femora pale; forefemur: tibiae: tarsi=1.0: 2.9: 1.9; femora of all legs with slightly pigmented apex; legs with paired claws, one hooked, one blunt. Wing pale, transparent, with brown veins, subcosta robust, yellow-brown (Figs. 9, 16). Genitalia: forceps covered with dense, tiny setae on surface; four spines located on the apex. Penis lobes fused, posterior margin with slight median emargination. Styliger plate pale yellow, with brown markings on the middle of the base (Figs. 10���11, 17��� 18). Caudal filaments pale. Female imago (in alcohol): Body length 2.6 mm, caudal filaments 1.3 mm. Similar to male. Etymology. The epithet pycnacantha is from the Latin pycnacanthus (dense), in reference to the surface of the forceps which are covered with dense, tiny setae. Diagnosis and discussion. To date only six Caenis species have been reported from Chinese Mainland (Eaton 1884; Klap��lek 1905; Ulmer 1936; Zhou et al. 1997, 2000; Gui et al. 1999; Zhou & Zheng 2004). C. pycnacantha sp. nov. is similar to C. nigropunctata (see Ulmer 1940) because both have three longitudinal pale stripes on larval abdominal terga 7���9, and the forceps have dense setae on the surface and spines on the apex. However, they can be separated by the following characters: 1. The larval legs of C. pycnacantha sp. nov. have obvious dark brown markings, which are absent in C. nigropunctata; 2. The larval abdomen of C. nigropunctata is slightly paler than in C. pycnacantha sp. nov., especially that of terga 10; 3. The apical segment of maxillary palpi of C. pycnacantha sp. nov. is slightly longer than second one, but those of C. nigropunctata are subequal; 4.The adult antennae of the new species are pale and the flagellum is slightly expanded at the base; in C. nigropunctata the antennal base is broader. Moreover, in the male imago of C. pycnacantha sp. nov., the forceps have four large spines at apex compared against C. nigropunctata with a tuft of spines on the same position. Kang & Yang (1994, 1996) and Tong & Dudgeon (2002) described totally 10 Caenis species from Chinese Taiwan and Hong Kong area, but all of them are reported from larval stages only and are distinguished by color pattern and/or mandibular seta number or length. It is believed here that those characters can vary in certain level. Without imaginal materials, it is very difficult to compare the new species here to these species. Malzacher (1984) divided the European Caenis species into two lineages based on forceps characteristics: horaria ���lineage (with straight, strongly sclerotised tips of forceps) and macrura -lineage (bent forceps with some bristles or a tuft of long spines on tips). The macrura -lineage is divided further into five species groups. C. pycnacantha sp. nov. can be placed into the macrura -lineage and macrura -group because of the apical spines on the forceps; it is distinguished from other members of the group by the dense setae on the surface of the forceps. Biology. Most nymphs of C. pycnacantha sp. nov. were collected from small pools in streams with sediments of thick leaves and soil. Nymphs were often found under surface of stones in water. This species can be found and collected in most seasons except cold winter (usually from end of November to February). The adults are easily reared from mature nymphal stage, those having dark wing pads can become adults in 2��� 3 days in the rearing tank. However, we did not find living adults during our daytime field collections or at light in the evening although some dead imagos were seen on the water surface during the day. Zhou et al. (2004) reported that in dawn the collect light can attract many caenid adults and subimagos. It is suspected the new species emerges in the early morning too. The width of streams during collection in Zijin Hill was about 1 m and depth of water was less than 40 cm (some places only 10 cm). The streams are often represented by small lentic pools and water flows under the stream bed in some sections. Except after rain, the streams flow slowly, less than 20 cm /s. There is a heavy tree canopy shading most of the stream (Fig. 19). Material examined. Holotype: 1 male, Ying-Tuo village, May 24, 2004, Peng-LI; Paratypes: 24 Nymphs, 6 males 20 females (reared from larvae in lab), same data as for holotype; 12 Nymphs, Ling-Gu-Si Temple, May 24, 2006, Peng-LI; 3 Nymphs, Water-side Pavilion, May 29, 2007, Dong LIU, Hui XIE and Shi-Lei WANG; 10 Nymphs, Ying-Tuo village, May 11, 2008, Shi-Lei WANG, Hui XIE, Yan-Yan JIA and Ping CHEN; 2 Nymphs, Ying-Tuo village, Sep. 22, 2008, Shi-Lei WANG, Yan-Yan JIA and Ping CHEN. All types and specimens are collected from Zijin Hill, Nanjing City, Jiangsu Province, China and deposited in the Mayfly Collection, College of Life Sciences, Nanjing Normal University, China., Published as part of Jia, Yan-Yan, Qin, Jia-Zhang, Ju, Min & Zhou, Chang-Fa, 2010, A new mayfly species of Caenis from headwater of Zijin Hill (Nanjing, Eastern China) (Ephemeroptera: Caenidae), pp. 61-68 in Zootaxa 2535 on pages 61-67, DOI: 10.5281/zenodo.196612, {"references":["Eaton, A. E. (1884) A revisional monograph of recent Ephemeridae or mayflies. Transactions of the Linnaeus Society of Londo n, Second series, Zoology, vol. 3, part 2, 77 - 152.","Klapalek, F. (1905) Plecopteren und Ephemeriden aus Java, gesammelt von Prof. K. Kraepelin 1904. Mitteilungen aus dem Naturhistorischen Museum in Hamburg, 22, 103 - 107.","Ulmer, G. (1936) Neue Chinesische Ephemeropteren, Nebst Ubersicht Uber die bisher aus China bekannten Arten. Peking Natural History Bulletin, 10 (3), 210 - 215.","Zhou, C. F., Gui, H. & Su, C. R. (1997) Three new species of Ephemeroptera (Insecta) from Henan Province of China. Entomotaxonomia, 19, 268 - 272 (in Chinese).","Zhou, C. F., Gui, H. & Sui, C. R. (2000) The first record of the genus Brachycercus in China with description of a new species (Ephemeroptera: Caenidae). Entomologia Sinica, 7, 132 - 134.","Zhou, C. F. & Zheng, L. Y. (2004) A Preliminary Study on the Genus Caenis (Ephemeroptera: Caenidae) form Chinese Mainland, with Description of a New Species. Entomotaxonomia, 26, 1 - 7.","Kang, S. C. & Yang, C. T. (1994) Caenidae of Taiwan (Ephemeroptera). Chinese Journal of Entomology, 14, 93 - 113.","Kang, S. C. & Yang, C. T. (1996) A new species of Caenis Stephens (Ephemeroptera: Caenidae) from Taiwan. Chinese Journal of Entomology, 16, 55 - 59.","Tong, X. L. & Dudgeon, D. (2002) Three new species of the genus Caenis from Hong Kong, China (Ephemeroptera: Caenidae). Journal of Zoological Research, 23 (3), 232 - 238.","Malzacher, P. (1984): Die europaischen Arten der Gattung Caenis Stephens (Insecta: Ephemeroptera). Stuttgarter Beitrage zur Naturkunde, Ser. A., 373 (48), 1 - 48."]}

Cincticostella bifurcata sp. nov. Mature larva: (in ethanol); body length 4.0–9.2 mm. Caudal filaments 3.0–5.5 mm. Body yellowish-brown. Ocelli, clypeus dorsal surface and vertex with various small tubercles and ridges (Figs. 1, 14). Mouthparts: anteromedian emargination of labrum shallow; dorsal labial surface with scattered setae and thick row of submarginal setae, free margins with relatively longer setae; ventral surface densely and irregularly covered with setae (Fig. 2). Mandible with dense row of long setae on dorsal and lateral surface; outer incisor apex trifurcated, inner incisor bifurcated; prostheca represented by tuft of long setae; right mandible with tuft of bristles on inner margin (Figs. 3, 4). Superlinguae of hypopharynx with row of setae on anterior margin, dorsal and ventral surfaces with thick setae; lingua with very sparse and tiny setae on dorsal and ventral surfaces (Fig. 5). Maxilla with two larger spines which ventral margin with several denticulate, four setae on inner margin, bushy golden and long hairs on apex and lateral surface, galea-lacinia with several setae on inner margin near base; maxillary palpi 2 -segmented, basal segment slightly broader, apex of 2 nd segment with constriction (Fig. 6). Labium with round glossae; glossae and paraglossae with setae on margins and dorsal surface; labial palpi 3 -segmented, basal segment length subequal to 2 nd segment, 2 nd segment with setae on margins and dorsal surface, apical segment with no setae and much thinner than other segments; mentum small, submentum with sparse setae and spines on margins, irregular short spines and setae on dorsal surface, but smooth on ventral surface (Fig. 7). Thorax: Pronotum expanded laterally, with anterolateral angles projecting anteriorly, mesonotum projections broad, thoracic nota with various irregular ridges and tubercles, pair of longitudinal ridges and sharp posterior projections between forewing pads (Figs. 1, 14). Femora of all legs flattened, especially those of mid- and hindfemora. Femora of all legs longer than tibiae, and tibiae longer than tarsi. Forefemora with short spines and thick setae on margin, foretibiae and foretarsi with setae on both outer and inner margins(Fig. 8). Midleg and hindleg similar: femora flatted, with longitudinal ridge on dorsal surface, some setae on basal outer margin; outer margin serrate; outer and inner margins with sparse setae near apex of tibiae, whole tarsi with setae on margins (Figs. 9, 10). Claws of all legs with 2 denticles, basal one bigger than distal one (Fig. 11). Abdomen: Pairs of tubercles present on abdominal terga 2-10, those on terga 2, 3, 4, 10 smaller than others; those on terga 5–8 strong, bifurcated apically (Figs. 12, 13); those on tergum 9 sharp-pointed but not bifurcated. Gills present on abdominal terga 3–7, gill 3 without medial transverse band of weakened membrane, dorsal lobe of gills 3–5 similar in shape, gill 6 diminished in size compared to gills 3–5, gill 7 very small and wholly covered by gill 6; ventral lamellae of all gills separated into several lobes, those of gills 3–5 further grouped into two parts, one clear cleft between them; those of gills 6–7 fused together, not bifurcate. Gill 7 located near posterolateral corner. Caudal filaments with tiny spines at articulations. Etymology: bifurcata (feminine), from Latin adjective bifurcatus (bifurcate), indicates tubercles of abdominal terga 5–8 are bifurcated apically on body of larva. Diagnosis: The new species is similar to C. femorata because of their significantly flattened bodies, legs and large projections on the pronotum and mesonotum. The new species can be distinguished from the latter by the following characters: (1) smaller body size; (2) body with many ridges and tubercles dorsally (Fig. 1); (3) mid- and hind femora with smaller serrated free margins (Figs. 9-10); (4) body and legs less flattened; (5) projections of thorax much smaller; (6) abdominal terga 5-8 with bifurcated tubercles(Fig. 13) instead of simple type as the C. femorata. Material examined: holotype: 1 larva (L), Zi-Ma river (29.00978°N, 102.28003°E, elev. 2175m), Conservation station of Zi-Ma river, Conservation area of Li-Zi-Ping, Shi-Mian County, Si-Chuan Province, July 1, 2005, Chang-Fa ZHOU. Paratypes: 2 L, same data as for holotype. 1 L, San-Gang (27.45°N, 117.40°E), Fu-Jian Province, June 7, 1987. 3 L, Mei-Zi river (26 ° 20.905 ΄ N, 100 °01.290΄ E, elev. 2319m, error 9m), Fu- Tian village, Niu-Jie countryside, Er-Yuan County, Da-Li State, Yun-Nan Province, July 8, 2008, Hui XIE, Yan-Yan JIA and Ping CHEN. All types deposited in the Mayfly Collection, College of Life Sciences, Nanjing Normal University, P. R. China 1, habitus of mature larva; 2, labrum (dorsum on left and venter on right); 3, left mandible; 4, right mandible; 5, hypopharynx (dorsum on left and venter on right); 6, right maxilla (ventral); 7, labium (dorsum on left and venter on right); 8, forleg; 9, midleg; 10, hindleg; 11, claw; 12, tubercle present on tergum 8.

Key to the known species of the genus Cincticostella (mature larva, larval stage of C. indica has not been reported so far) 1. Middle and hind femora expanded and usually somewhat serrate (figs. 1, 17, 20, 22, 23)......................................... 2 1 ���. Middle and hind femora not significantly expanded (figs. 19, 21, 24, 25)................................................................... 5 2. Abdominal tergal spines on segments 5-8 bifurcate (figs. 1, 12, 13, 22) ........................................ C. bifurcata sp.nov. 2 ���. Abdominal tergal spines on 5-8 not bifurcate............................................................................................................... 3 3. Midfemur with deep, U-shaped, apical notch at joint with tibia (fig. 23).................................................................... 4 3 ���. Midfemur without such notch (fig. 28)........................................................................................................ C. braaschi 4. Pronotum greatly expanded around head capsule (figs. 17, 23) .................................................................. C. femorata 4 ���. Pronotum not extremely expanded (figs. 20, 26) ............................................................................................ C. insolta 5. Maxillary palp present, sometimes vestigial (see figs. 6, 16)....................................................................................... 6 5 ���. Maxillary palp absent......................................................................................................................................... C. gosei 6. Maxilla with crown expanded and flattened (fig. 16)................................................................................................... 7 6 ���. Maxilla with crown only slightly expanded and not flattened...................................................................................... 8 22, C. bifurcata sp. nov.; 23, C. femorata; 24, C. fusca; 25, C. gosei; 26, C. insolta; 27, C. nigra. 7. Mesal plate with paired ridges or projections on posterior margin; median filament length subequal to length of abdomen ...................................................................................................................................................... C. orientalis 7 ���. Mesal plate without such ridges or projections; median filament length much greater than length of abdomen.......... .................................................................................................................................................................. C. corpulenta 8. Anterolateral projections of mesothorax notched medially, sometimes with sharply pointed appearance (fig. 15).... 9 8 ���. Anterolateral projections of mesothorax blunt and not notched, with margins usually subparallel to lateral aspect of body............................................................................................................................................................................. 11 9. Abdominal terga with paired spines on posterior margins......................................................................................... 10 9 ���. Abdominal terga without such spines ..................................................................................... C. szechuanensis sp.nov. 10. Setae on lateral margins of abdominal segments numerically reduced, stout and spinelike; longitudinal stripes absent from abdominal terga; and median filament length nearly subequal to length of abdomen .............................. C. fusca 10 ���. Setae on lateral margins of abdominal segments numerous and generally spatulate; distinct longitudinal stripes usu- ally present on abdominal terga; and median filament length greater than length of abdomen ............. C. levanidovae 11. Genae expanded and angulate ......................................................................................................................... C.colossa 11 ���. Genae rounded............................................................................................................................................................ 12 12. Upper surface of femora densely covered with short, clavate setae .......................................................... C.elongatula 12 ���. Upper surface of femora smooth, without short, clavate setae .......................................................................... C. nigra, Published as part of Xie, Hui, Jia, Yan-Yan, Chen, Ping, Jacobus, Luke M. & Zhou, Chang-Fa, 2009, Two new Cincticostella species from China with a larval key to species of the genus (Ephemeroptera: Ephemerellidae), pp. 53-61 in Zootaxa 2299 on pages 58-60, DOI: 10.5281/zenodo.191662

Cincticostella bifurcata sp. nov. Mature larva: (in ethanol); body length 4.0���9.2 mm. Caudal filaments 3.0���5.5 mm. Body yellowish-brown. Ocelli, clypeus dorsal surface and vertex with various small tubercles and ridges (Figs. 1, 14). Mouthparts: anteromedian emargination of labrum shallow; dorsal labial surface with scattered setae and thick row of submarginal setae, free margins with relatively longer setae; ventral surface densely and irregularly covered with setae (Fig. 2). Mandible with dense row of long setae on dorsal and lateral surface; outer incisor apex trifurcated, inner incisor bifurcated; prostheca represented by tuft of long setae; right mandible with tuft of bristles on inner margin (Figs. 3, 4). Superlinguae of hypopharynx with row of setae on anterior margin, dorsal and ventral surfaces with thick setae; lingua with very sparse and tiny setae on dorsal and ventral surfaces (Fig. 5). Maxilla with two larger spines which ventral margin with several denticulate, four setae on inner margin, bushy golden and long hairs on apex and lateral surface, galea-lacinia with several setae on inner margin near base; maxillary palpi 2 -segmented, basal segment slightly broader, apex of 2 nd segment with constriction (Fig. 6). Labium with round glossae; glossae and paraglossae with setae on margins and dorsal surface; labial palpi 3 -segmented, basal segment length subequal to 2 nd segment, 2 nd segment with setae on margins and dorsal surface, apical segment with no setae and much thinner than other segments; mentum small, submentum with sparse setae and spines on margins, irregular short spines and setae on dorsal surface, but smooth on ventral surface (Fig. 7). Thorax: Pronotum expanded laterally, with anterolateral angles projecting anteriorly, mesonotum projections broad, thoracic nota with various irregular ridges and tubercles, pair of longitudinal ridges and sharp posterior projections between forewing pads (Figs. 1, 14). Femora of all legs flattened, especially those of mid- and hindfemora. Femora of all legs longer than tibiae, and tibiae longer than tarsi. Forefemora with short spines and thick setae on margin, foretibiae and foretarsi with setae on both outer and inner margins(Fig. 8). Midleg and hindleg similar: femora flatted, with longitudinal ridge on dorsal surface, some setae on basal outer margin; outer margin serrate; outer and inner margins with sparse setae near apex of tibiae, whole tarsi with setae on margins (Figs. 9, 10). Claws of all legs with 2 denticles, basal one bigger than distal one (Fig. 11). Abdomen: Pairs of tubercles present on abdominal terga 2-10, those on terga 2, 3, 4, 10 smaller than others; those on terga 5���8 strong, bifurcated apically (Figs. 12, 13); those on tergum 9 sharp-pointed but not bifurcated. Gills present on abdominal terga 3���7, gill 3 without medial transverse band of weakened membrane, dorsal lobe of gills 3���5 similar in shape, gill 6 diminished in size compared to gills 3���5, gill 7 very small and wholly covered by gill 6; ventral lamellae of all gills separated into several lobes, those of gills 3���5 further grouped into two parts, one clear cleft between them; those of gills 6���7 fused together, not bifurcate. Gill 7 located near posterolateral corner. Caudal filaments with tiny spines at articulations. Etymology: bifurcata (feminine), from Latin adjective bifurcatus (bifurcate), indicates tubercles of abdominal terga 5���8 are bifurcated apically on body of larva. Diagnosis: The new species is similar to C. femorata because of their significantly flattened bodies, legs and large projections on the pronotum and mesonotum. The new species can be distinguished from the latter by the following characters: (1) smaller body size; (2) body with many ridges and tubercles dorsally (Fig. 1); (3) mid- and hind femora with smaller serrated free margins (Figs. 9-10); (4) body and legs less flattened; (5) projections of thorax much smaller; (6) abdominal terga 5-8 with bifurcated tubercles(Fig. 13) instead of simple type as the C. femorata. Material examined: holotype: 1 larva (L), Zi-Ma river (29.00978��N, 102.28003��E, elev. 2175m), Conservation station of Zi-Ma river, Conservation area of Li-Zi-Ping, Shi-Mian County, Si-Chuan Province, July 1, 2005, Chang-Fa ZHOU. Paratypes: 2 L, same data as for holotype. 1 L, San-Gang (27.45��N, 117.40��E), Fu-Jian Province, June 7, 1987. 3 L, Mei-Zi river (26 �� 20.905 �� N, 100 ��01.290�� E, elev. 2319m, error 9m), Fu- Tian village, Niu-Jie countryside, Er-Yuan County, Da-Li State, Yun-Nan Province, July 8, 2008, Hui XIE, Yan-Yan JIA and Ping CHEN. All types deposited in the Mayfly Collection, College of Life Sciences, Nanjing Normal University, P. R. China 1, habitus of mature larva; 2, labrum (dorsum on left and venter on right); 3, left mandible; 4, right mandible; 5, hypopharynx (dorsum on left and venter on right); 6, right maxilla (ventral); 7, labium (dorsum on left and venter on right); 8, forleg; 9, midleg; 10, hindleg; 11, claw; 12, tubercle present on tergum 8., Published as part of Xie, Hui, Jia, Yan-Yan, Chen, Ping, Jacobus, Luke M. & Zhou, Chang-Fa, 2009, Two new Cincticostella species from China with a larval key to species of the genus (Ephemeroptera: Ephemerellidae), pp. 53-61 in Zootaxa 2299 on pages 54-55, DOI: 10.5281/zenodo.191662

Key to the known species of the genus Cincticostella (mature larva, larval stage of C. indica has not been reported so far) 1. Middle and hind femora expanded and usually somewhat serrate (figs. 1, 17, 20, 22, 23)......................................... 2 1 ’. Middle and hind femora not significantly expanded (figs. 19, 21, 24, 25)................................................................... 5 2. Abdominal tergal spines on segments 5-8 bifurcate (figs. 1, 12, 13, 22) ........................................ C. bifurcata sp.nov. 2 ’. Abdominal tergal spines on 5-8 not bifurcate............................................................................................................... 3 3. Midfemur with deep, U-shaped, apical notch at joint with tibia (fig. 23).................................................................... 4 3 ’. Midfemur without such notch (fig. 28)........................................................................................................ C. braaschi 4. Pronotum greatly expanded around head capsule (figs. 17, 23) .................................................................. C. femorata 4 ’. Pronotum not extremely expanded (figs. 20, 26) ............................................................................................ C. insolta 5. Maxillary palp present, sometimes vestigial (see figs. 6, 16)....................................................................................... 6 5 ’. Maxillary palp absent......................................................................................................................................... C. gosei 6. Maxilla with crown expanded and flattened (fig. 16)................................................................................................... 7 6 ’. Maxilla with crown only slightly expanded and not flattened...................................................................................... 8 22, C. bifurcata sp. nov.; 23, C. femorata; 24, C. fusca; 25, C. gosei; 26, C. insolta; 27, C. nigra. 7. Mesal plate with paired ridges or projections on posterior margin; median filament length subequal to length of abdomen ...................................................................................................................................................... C. orientalis 7 ’. Mesal plate without such ridges or projections; median filament length much greater than length of abdomen.......... .................................................................................................................................................................. C. corpulenta 8. Anterolateral projections of mesothorax notched medially, sometimes with sharply pointed appearance (fig. 15).... 9 8 ’. Anterolateral projections of mesothorax blunt and not notched, with margins usually subparallel to lateral aspect of body............................................................................................................................................................................. 11 9. Abdominal terga with paired spines on posterior margins......................................................................................... 10 9 ’. Abdominal terga without such spines ..................................................................................... C. szechuanensis sp.nov. 10. Setae on lateral margins of abdominal segments numerically reduced, stout and spinelike; longitudinal stripes absent from abdominal terga; and median filament length nearly subequal to length of abdomen .............................. C. fusca 10 ’. Setae on lateral margins of abdominal segments numerous and generally spatulate; distinct longitudinal stripes usu- ally present on abdominal terga; and median filament length greater than length of abdomen ............. C. levanidovae 11. Genae expanded and angulate ......................................................................................................................... C.colossa 11 ’. Genae rounded............................................................................................................................................................ 12 12. Upper surface of femora densely covered with short, clavate setae .......................................................... C.elongatula 12 ’. Upper surface of femora smooth, without short, clavate setae .......................................................................... C. nigra

Xie, Hui, Jia, Yan-Yan, Chen, Ping, Jacobus, Luke M., Zhou, Chang-Fa (2009): Two new Cincticostella species from China with a larval key to species of the genus (Ephemeroptera: Ephemerellidae). Zootaxa 2299: 53-61, DOI: 10.5281/zenodo.191662