[Cross-sectional associations between reproductive lifespan duration and urinary albumin-creatinine ratio].

Objective: To investigate the association between reproductive lifespan duration (RLD) and urinary albumin-creatinine ratio (UACR) in a Chinese postmenopausal population. Methods: This cross-sectional study included 11 055 naturally postmenopausal women from seven regions of China from May to December 2011. RLD was divided into four groups. Propensity score matching was performed to reduce bias, and logistic regressions and stratifications were conducted to investigate the association between RLD and increased UACR (≥30 mg/g). Mediation effect analysis was performed to quantify the effect of RLD on cardiovascular disease (CVD) induced by elevated UACR. Results: There were 2 373 participants with a RLD of 18-31 years, 2 888 participants with a RLD of 32-34 years, 2 472 participants with a RLD of 35-36 years, and 3 322 participants with a RLD of 37-50 years. The shortest RLD (18-31 years) group was characterized with older age (P<0.001), a higher incidence of CVD (P=0.025), and the highest level of UACR (P<0.001). After adjusting for confounders, women with a longer RLD (37-50 years group) exhibited a lower risk of UACR elevation compared with those with the shortest RLD (18-31 years group) (OR=0.72, 95%CI 0.64-0.82, P<0.001). Every 1-year extension in RLD was linked to a 2% reduction in the risk of UACR elevation (OR=0.98, 95%CI 0.97-0.99, P<0.001). Stratified analysis revealed a more significant association between RLD and UACR in women who were a normal weight (P=0.003) or overweight (P=0.001), in those without CVD history (P=0.001), and in those with impaired estimated glomerular filtration rate (P=0.004). The mediation casual analysis showed that 3.0% of proteinuria inducing CVD events was mediated by RLD (P=0.048). Conclusion: A longer RLD (37-50 years) is associated with a lower UACR in Chinese postmenopausal women.

[Analysis of FBN1 genemutations in a pedigree with Marfan syndrome].

Mutations in fibrillin-1 (FBN1) were detected in an autosomal dominant Marfan syndrome (MFS) pedigree. The related phenotypes and the significance of mutation screening were discussed. Complete medical and cardiovascular examinations for all pedigree members were performed. Whole exons sequencing (WES) was used to sequence the DNA of the patients and their relatives. The potential pathogenic mutation sites were screened by bioinformatics method. Sanger sequencing was used to verify the mutation sites in the pedigree. The results showed that FBN1 missense mutation was c.6806 T>C in exon 56, resulting in isoleucine being replaced by threonine (p. Ile2269Thr). This mutation has not been reported in Chinese Han population. The occurrence of the mutations strongly correlated with the phenotypes of the patients. The results expand the mutation spectrum of FBN1, and it is helpful to further explore the molecular pathogenesis of MFS and MFS related diseases.

Observation of Weyl Nodes in Robust Type-II Weyl Semimetal WP_{2}.

Distinct to type-I Weyl semimetals (WSMs) that host quasiparticles described by the Weyl equation, the energy dispersion of quasiparticles in type-II WSMs violates Lorentz invariance and the Weyl cones in the momentum space are tilted. Since it was proposed that type-II Weyl fermions could emerge from (W,Mo)Te_{2} and (W,Mo)P_{2} families of materials, a large number of experiments have been dedicated to unveiling the possible manifestation of type-II WSMs, e.g., surface-state Fermi arcs. However, the interpretations of the experimental results are very controversial. Here, using angle-resolved photoemission spectroscopy supported by the first-principles calculations, we probe the tilted Weyl cone bands in the bulk electronic structure of WP_{2} directly, which are at the origin of Fermi arcs at the surfaces and transport properties related to the chiral anomaly in type-II WSMs. Our results ascertain that, due to the spin-orbit coupling, the Weyl nodes originate from the splitting of fourfold degenerate band-crossing points with Chern numbers C=±2 induced by the crystal symmetries of WP_{2}, which is unique among all the discovered WSMs. Our finding also provides a guiding line to observe the chiral anomaly that could manifest in novel transport properties.

Distinct Evolutions of Weyl Fermion Quasiparticles and Fermi Arcs with Bulk Band Topology in Weyl Semimetals.

The Weyl semimetal phase is a recently discovered topological quantum state of matter characterized by the presence of topologically protected degeneracies near the Fermi level. These degeneracies are the source of exotic phenomena, including the realization of chiral Weyl fermions as quasiparticles in the bulk and the formation of Fermi arc states on the surfaces. Here, we demonstrate that these two key signatures show distinct evolutions with the bulk band topology by performing angle-resolved photoemission spectroscopy, supported by first-principles calculations, on transition-metal monophosphides. While Weyl fermion quasiparticles exist only when the chemical potential is located between two saddle points of the Weyl cone features, the Fermi arc states extend in a larger energy scale and are robust across the bulk Lifshitz transitions associated with the recombination of two nontrivial Fermi surfaces enclosing one Weyl point into a single trivial Fermi surface enclosing two Weyl points of opposite chirality. Therefore, in some systems (e.g., NbP), topological Fermi arc states are preserved even if Weyl fermion quasiparticles are absent in the bulk. Our findings not only provide insight into the relationship between the exotic physical phenomena and the intrinsic bulk band topology in Weyl semimetals, but also resolve the apparent puzzle of the different magnetotransport properties observed in TaAs, TaP, and NbP, where the Fermi arc states are similar.

[Effects of interleukin-4-modified gold nanozymes on the full-thickness skin defects in diabetic mice].

Objective: To investigate the effects and mechanism of interleukin-4-modified gold nanoparticle (IL-4-AuNP) on the wound healing of full-thickness skin defects in diabetic mice. Methods: Experimental research methods were adopted. Gold nanoparticle (AuNP) and IL-4-AuNP were synthesized by improving the methods described in published literature. The morphology of those two particles were photographed by transmission electron microscopy, and their particle sizes were calculated. The surface potential and hydration particle size of the two particles were detected by nanoparticle potentiometer and particle size analyzer, respectively. The clearance rate of IL-4-AuNP to hydrogen peroxide and superoxide anion was measured by hydrogen peroxide and superoxide anion kits, respectively. Mouse fibroblast line 3T3 cells were used and divided into the following groups by the random number table (the same below): blank control group, hydrogen peroxide alone group treated with hydrogen peroxide only, hydrogen peroxide+IL-4-AuNP group treated with IL-4-AuNP for 0.5 h and then treated with hydrogen peroxide. After 24 h of culture, the reactive oxygen species (ROS) levels of cells were detected by immunofluorescence method; cell count kit 8 was used to detect relative cell survival rate. The macrophage Raw264.7 mouse cells were then used and divided into blank control group and IL-4-AuNP group that treated with IL-4-AuNP. After 24 h of culture, the expression of arginase 1 (Arg-1) in cells was observed by immunofluorescence method. Twelve male BALB/c mice (mouse age, sex, and strain, the same below) aged 8 to 10 weeks were divided into IL-4-AuNP group and blank control group, treated accordingly. On the 16th day of treatment, whole blood samples were collected from mice for analysis of white blood cell count (WBC), red blood cell count (RBC), hemoglobin level, or platelet count and the level of aspartate aminotransferase (AST), alanine transaminase (ALT), urea, or creatinine. The inflammation, bleeding, or necrosis in the heart, liver, spleen, lung, and kidney tissue of mice were detected by hematoxylin-eosin (HE). Another 36 mice were selected to make diabetic model, and the full-thickness skin defect wounds were made on the back of these mice. The wounds were divided into blank control group, AuNP alone group, and IL-4-AuNP group, with 12 mice in each group, and treated accordingly. On the 0 (immediately), 4th, 9th, and 15th day of treatment, the wound condition was observed and the wound area was calculated. On the 9th day of treatment, HE staining was used to detect the length of neonatal epithelium and the thickness of granulation tissue in the wound. On the 15th day of treatment, immunofluorescence method was used to detect ROS level and the number of Arg-1 positive cells in the wound tissue. The number of samples was 6 in all cases. Data were statistically analyzed with independent sample t test, corrected t test, Tukey test, or Dunnett T3 test. Results: The size of prepared AuNP and IL-4-AuNP were uniform. The particle size, surface potential, and hydration particle size of AuNP and IL-4-AuNP were (13.0±2.1) and (13.9±2.5) nm, (-45.8±3.2) and (-20.3±2.2) mV, (14±3) and (16±4) nm, respectively. For IL-4-AuNP, the clearance rate to hydrogen peroxide and superoxide anion were (69±4)% and (52±5)%, respectively. After 24 h of culture, the ROS level of 3T3 in hydrogen peroxide alone group was significantly higher than that in blank control group (q=26.12, P<0.05); the ROS level of hydrogen peroxide+IL-4-AuNP group was significantly lower than that in hydrogen peroxide alone group (q=25.12, P<0.05) and close to that in blank control group (P>0.05). After 24 h of culture, the relative survival rate of 3T3 cells in hydrogen peroxide+IL-4-AuNP group was significantly higher than that in hydrogen peroxide alone group (t=51.44, P<0.05). After 24 h of culture, Arg-1 expression of Raw264.7 cells in IL-4-AuNP group was significantly higher than that in blank control group (t'=8.83, P<0.05).On the 16th day of treatment, there were no significant statistically differences in WBC, RBC, hemoglobin level, or platelet count and the level of AST, ALT, urea, or creatinine of mice between blank control group and IL-4-AuNP group (P>0.05). No obvious inflammation, bleeding or necrosis was observed in the heart, liver, spleen, lung, and kidney of important organs in IL-4-AuNP group, and no significant changes were observed compared with blank control group. On the 0 and 4th day of treatment, the wound area of diabetic mice in blank control group, AuNP alone group, and IL-4-AuNP group had no significant difference (P>0.05). On the 9th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 9.45 and 14.87, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=5.42, P<0.05). On the 15th day of treatment, the wound areas both in AuNP alone group and IL-4-AuNP group were significantly smaller than that in blank control group (with q values of 4.84 and 20.64, respectively, P<0.05), the wound area in IL-4-AuNP group was significantly smaller than that in AuNP alone group (q=15.80, P<0.05); moreover, inflammations such as redness and swelling were significantly reduced in IL-4-AuNP group compared with the other two groups. On the 9th day of treatment, compared with blank control group and AuNP alone group, the length of neonatal epithelium in the wound of diabetic mice in IL-4-AuNP group was significantly longer (all P<0.05), and the thickness of the granulation tissue in the wound was significantly increased (with q values of 11.33 and 9.65, respectively, all P<0.05). On the 15th day of treatment, compared with blank control group, ROS levels in wound tissue of diabetic mice in AuNP alone group and IL-4-AuNP group were significantly decreased (P<0.05). On the 15th day of treatment, the number of Arg-1 positive cells in the wounds of diabetic mice in IL-4-AuNP group was significantly more than that in blank control group and AuNP alone group, respectively (all P<0.05). Conclusions: IL-4-AuNP is safe in vivo, and can improve the oxidative microenvironment by removing ROS and induce macrophage polarization towards M2 phenotype, thus promote efficient diabetic wound healing and regeneration of full-thickness skin defects in diabetic mice.

Observation of a singular Weyl point surrounded by charged nodal walls in PtGa.

Constrained by the Nielsen-Ninomiya no-go theorem, in all so-far experimentally determined Weyl semimetals (WSMs) the Weyl points (WPs) always appear in pairs in the momentum space with no exception. As a consequence, Fermi arcs occur on surfaces which connect the projections of the WPs with opposite chiral charges. However, this situation can be circumvented in the case of unpaired WP, without relevant surface Fermi arc connecting its surface projection, appearing singularly, while its Berry curvature field is absorbed by nontrivial charged nodal walls. Here, combining angle-resolved photoemission spectroscopy with density functional theory calculations, we show experimentally that a singular Weyl point emerges in PtGa at the center of the Brillouin zone (BZ), which is surrounded by closed Weyl nodal walls located at the BZ boundaries and there is no Fermi arc connecting its surface projection. Our results reveal that nontrivial band crossings of different dimensionalities can emerge concomitantly in condensed matter, while their coexistence ensures the net topological charge of different dimensional topological objects to be zero. Our observation extends the applicable range of the original Nielsen-Ninomiya no-go theorem which was derived from zero dimensional paired WPs with opposite chirality.

Spin fluctuation induced Weyl semimetal state in the paramagnetic phase of EuCd2As2.

Weyl fermions as emergent quasiparticles can arise in Weyl semimetals (WSMs) in which the energy bands are nondegenerate, resulting from inversion or time-reversal symmetry breaking. Nevertheless, experimental evidence for magnetically induced WSMs is scarce. Here, using photoemission spectroscopy, we observe that the degeneracy of Bloch bands is already lifted in the paramagnetic phase of EuCd2As2. We attribute this effect to the itinerant electrons experiencing quasi-static and quasi-long-range ferromagnetic fluctuations. Moreover, the spin-nondegenerate band structure harbors a pair of ideal Weyl nodes near the Fermi level. Hence, we show that long-range magnetic order and the spontaneous breaking of time-reversal symmetry are not essential requirements for WSM states in centrosymmetric systems and that WSM states can emerge in a wider range of condensed matter systems than previously thought.

Chronic subdural hematoma management: clarifying the definitions of outcome measures to better understand treatment efficacy - a systematic review and meta-analysis.

OBJECTIVE: A long history of inconsistencies in the definitions of the outcome measures for chronic subdural hematomas (CSDHs) has contributed to the controversy over the optimal surgical strategy for CSDH treatment. Clarifying these definitions, reassess the available data, and systematically review the prior literature may provide better insight into the differences in treatment efficacy for CSDH. MATERIALS AND METHODS: The clinical course of CSDH was described with a series of strictly defined outcome measures. PubMed, Cochrane Library, and ScienceDirect databases were searched for comparative studies of two main surgical techniques for CSDH, including burr hole craniotomy (BHC) and twist drill craniotomy (TDC). Data were collected with uniform criteria and analyzed using a random-effects model to estimate the mortality, recurrence, operative failure, and cure rates of each treatment. RESULTS: Twelve comparative studies that examined 2,027 CSDH patients were included. The analysis results indicated that TDC and BHC treatments were similar in the mortality rates (RR, 1.25; 95% CI, 0.83-1.87; I2 = 0%; p = 0.28) and the recurrence rates (RR, 1.29; 95% CI, 0.87-1.92; I2 = 13%; p = 0.21) for CSDH patients. However, TDC had a significantly higher operative failure rate compared with BHC (RR, 0.35; 95% CI, 0.15-0.83; I2 = 0%; p = 0.02), whereas patients treated by a TDC approach tended to achieve higher cure rates compared with BHC (RR, 0.92; 95% CI, 0.86-0.99; I2 = 55%; p = 0.02). CONCLUSIONS: The clarification of the definitions related to CSDH outcome facilitates the interpretation of differences in treatment efficacy. The TDC approach manifested a significantly higher operative failure rate compared with the BHC approach; however, TDC showed a tendency in achieving a long-term neurologic cure.

Effects of mebendazole, albendazole, and praziquantel on glutathione S-transferase and superoxide dismutase of Echinococcus granulosus cyst wall harbored in mice.

AIM: To explore the existence of glutathione S-transferase (GST) and superoxide dismutase (SOD) in Echinococcus granulosus cyst, and the effect of anthydatid drugs on these 2 enzymes, mice infected with E granulosus protoscoleces for 10-12 months were used. METHODS: The activity of GST was measured by conjugation rate of 1-chloro-2,4-dinitrobenzene with glutathione (reduced form), while the activity of SOD was measured by a modified autoxidation of 1, 2, 3-trihydroxy-benzene method. RESULTS: Activities of both GST and SOD in the cyst wall were 12.3 +/- 4.3 to 13.5 +/- 4.8 mumol min-1/mg protein and 4.4 +/- 2.9 to 6.1 +/- 1.4 U min-1/mg protein, respectively. When infected mice were treated ig with mebendazole (Meb) 25 mg kg-1 d-1 for 14 d, the GST activity of both collapsed and full cyst walls were inhibited by 30.1% and 26.8%, respectively. Whereas SOD activity of the cyst walls were activated by 105%-163%. Albendazole 300 mg kg-1 d-1 for 14 d and praziquantel 500 mg kg-1 d-1 for 14 d had no apparent effect on both GST and SOD of E granulosus cyst wall. CONCLUSION: The results suggested that the inhibition of GST activity in the cyst wall induced by Meb might damage the defence system of the parasite.

Effect of mebendazole on free amino acid composition of cyst wall and cyst fluid of Echinococcus granulosus harbored in mice.

Eighteen and 23 FAA components were detected in the cyst wall and cyst fluid of E granulosus, respectively, by using automatic amino acid analyzer. The concentrations of most of the determined FAA were higher in the cyst fluid than those in the cyst wall, especially the taurine was 5-fold higher. Mebendazole treatment resulted in an increase in the concentration of alanine, valine, lysine, and taurine in both cyst wall and cyst fluid, the most notable being the alanine in the cyst wall. The results are interpreted as a coupling of glycolysis and amino acid metabolism, suggesting an involvement of FAA metabolism in the mechanism of Meb action.

Effect of mebendazole and praziquantel on glucosephosphate isomerase and glyceraldehydephosphate dehydrogenase in Echinococcus granulosus cyst wall harbored in mice.

AIM: To study effects of antihydatid drugs on glucosephosphate isomerase (GPI) and glyceraldehydephosphate dehydrogenase (GAPDH) in Echinococcus granulosus cyst wall. METHODS: Mice infected with the parasite for 8-10 months were treated i.g. with mebendazole (Meb) or praziquantel (Pra). The activities of GPI and GAPDH in the cysts were measured by the formation of NADH or NADPH. RESULTS: GPI activity in the cyst wall was 197 +/- 103 U, while that of GAPDH was 25 +/- 13 U. When infected mice were treated i.g. with Meb 25-50 mg.kg-1.d-1 for 7-14 d, no apparent effect on the GAPDH activity in the cyst was found. In mice treated i.g. with praziquantel (Pra) 500 mg.kg-1.d-1 for 14 d, the GAPDH activity in the cyst wall was inhibited by 26.5%. As to GPI activity only the group treated i.g. with Meb 25 mg.kg-1.d-1 for 14 d showed 33.2% inhibition of the enzyme in the collapsed cyst wall. CONCLUSION: GPI and GAPDH are not the major targets attacked by the antihydatid drug.

Effects of mebendazole, albendazole, and praziquantel on fumarate hydratase, pyruvate kinase, and phosphoenolpyruvate carboxykinase of Echinococcus granulosus cyst wall harbored in mice.

Echinococcus granulosus cyst wall exhibited activities of fumarate hydratase (FH), pyruvate kinase (PK), and phosphoenolpyruvate carboxykinase (PEPCK) with 911-1433, 151-215, and 54-98U, respectively. The ratio of PK/PEPCK was 2.2-2.7, indicating that glycolysis is the main pathway of carbohydrate metabolism in the cyst wall. When infected mice were treated ig with mebendazole, albendazole or praziquantel at the respective daily dose of 25-50, 300, and 500 mg.kg-1 for 7-14 d, no apparent change of FH activity in the cyst wall was found, while PK and PEPCK activities in the cyst wall were markedly inhibited by mebendazole and albendazole. The inhibition rates of PK and PEPCK activities in mebendazole group were 85-88% and 90-92%, respectively, while in albendazole group were 55.3 and 71.6%, respectively. The results suggest that PK or PEPCK in the cyst wall may the important site attacked by effective anti-hydatid drugs.

[Effects of mebendazole on glucose, glycogen, lactic acid, and lactate dehydrogenase in Echinococcus granulosus cyst wall].

The glucose, glycogen, and lactic acid contents in Echinococcus granulosus cyst wall were 3570 +/- 623, 1276 +/- 126, and 267 +/- 29 micrograms.g-1, respectively. The lactate dehydrogenase (LDH) activity in the cyst wall was 4307 +/- 297 U/(min.mg protein). When infected mice were treated ig with mebendazole (Meb) 25 or 50 mg.kg-1.d-1 for 7-14 d, the glucose and glycogen contents in the cyst wall decreased markedly. No apparent changes in lactic acid and LDH activity were seen. The results suggested that Meb had no effect on glycolysis of the cyst wall, but it exhibited an inhibitory effect on the transport of exogenous glucose from the host to the cyst wall, which might further promote the decomposition of endogenous glycogen for energy supply and resulted in a decrease of endogenous glycogen content in the cyst wall.

Effects of mebendazole, albendazole, and praziquantel on succinate dehydrogenase, fumarate reductase, and malate dehydrogenase in Echinococcus granulosus cysts harbored in mice.

Echinococcus granulosus cyst wall possess high biochemical activities of malate dehydrogenase (MD) and fumarate reductase (FR), but low activity of succinate dehydrogenase (SD), suggesting that the cyst wall may utilize a partial reverse tricarboxylic acid cycle. When infected mice were given intragastrically with mebendazole, 25-50 mg.kg-1.d-1, albendazole 300 mg.kg-1.d-1 or praziquantel 500 mg.kg-1.d-1 for 7-14 d, no apparent effects on SD and FR activities of the cyst wall were found, while the MD activity was suppressed by all the 3 drugs, the inhibition rates being 34.6-61.6%, 59.8%, and 50.6%, respectively. The results suggested that MD may not be an important target for the antihydatidosis drugs.

Effect of artemether on glucose uptake and glycogen content in Schistosoma japonicum.

AIM: To study the effect of artemether (Art) on glucose uptake and glycogen content in schistosomes. METHODS: Schistosomes recovered from mice treated intragastrically with Art 300 mg.kg-1 for 24-48 h, were incubated in the drug-free medium containing [U-14C]glucose 11.1 MBq.L-1. The glycogen content, [U-14C]glucose uptake, and incorporation of [U-14C]glucose into worm glycogen in both male and female worms were determined. RESULTS: When above-mentioned schistosomes were exposed to drug-free medium containing [U-14C]glucose for 1-24 h, the glycogen contents of male and female worms decreased 27%-61% and 39%-78%, respectively. Only 3 out of 6 male worm groups showed 23%-35% decrease in glucose uptake, while much less glucose uptake was found in female worms in all groups with reduction rates of 18%-38%. Apart from 2 male groups no apparent change in the incorporation of [U-14C]glucose into the worm glycogen was seen. CONCLUSIONS: Art-induced glycogen reduction in schistosomes was related to an inhibition of glycolysis rather than an interference with glucose uptake.

Effect of artemether on glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, and pyruvate kinase of Schistosoma japonicum harbored in mice.

AIM: To study the effect of artemether (Art) on glyceraldehyde-phosphate dehydrogenase (GAPDH), phosphoglycerate kinase (PGK), and pyruvate kinase (PK) of S japanicum. METHODS: Mice infected with schistosome cercariae for 32-38 d were treated ig with Art 100-300 mg.kg-1 and killed 24-72 h after medication for collection of schistosomes. The activities of GAPDH, PGK, and PK of the worms were determined by measuring the formation of NADH or consumption of NAD. The lactate content of the worms was also measured. RESULTS: After the infected mice were treated ig with Art 300 mg.kg-1 for 24 h, the inhibition rates of GAPDH were 13% (Male) and 21% (Female), and 48 h later the inhibition rates of the enzyme were 6% (Male) and 28% (Female). When Art 300 mg.kg-1 was given to infected mice for 24 h and 48 h, the inhibition rates of PGK were 60% (Male) and 48% (Female) as well as 75% (Male) and 62% (Female), respectively. Similar results were seen in PK activity. At 72 h after treatment the reduction rate of lactate content in Female worm was 72%, while that of Male was 48%. CONCLUSION: In the glycolytic pathway of both Male and Female schistosomes, PGK and PK activities were inhibited by Art. The GAPDH activity of Female worms was also susceptible to Art, While that of Male worms showed only temporary inhibition after treatment with Art. The Art reduced lactate content more in Female than in Male worms.

[Effect of mebendazole on glucose uptake of Echinococcus granulosus cysts].

Mice infected with secondary cysts of Echinococcus granulosus were treated ig with mebendazole (Meb) 25 mg.kg-1.d-1 for 7-14 d. At 24 h after the last dose the endocysts in the treated mice were removed out for in vitro cultivation and exposed to [U-14C]glucose 11.1 kBq.ml-1 for 2 min, no apparent difference in radioactivity content in the cyst walls between the treated and control groups was observed. When [U-14C]glucose was given iv to the infected mice 24 h after they had been treated ig with Meb 25 mg.kg-1 or 50 mg.kg-1 daily for 14 d, the radioactivity content in the cyst wall and cyst fluid decreased significantly as compared to the corresponding control group. Nevertheless, no apparent change in the incorporation of radioactivity into the endogenous glycogen of the parasites was observed, although the glycogen in the cyst wall decreased markedly.