Ashwagandha and Its Active Ingredient, Withanolide A, Increase Phosphorylation of TrkB in Cultured Hippocampal Neurons

Aims: To primary rat embryonic hippocampal neurons in culture, ashwagandha or one of its active ingredients, withanolide A were added in the presence or absence of nutrient supplementation and then assayed for activity of the BDNF receptor, TrkB. Study Design: Primary hippocampal neurons were cultured and grown in nutrient-rich or nutrient-poor medium. Ashwagandha or withanolide A were then be added to both types of media with or without an inhibitor of TrkB or either the PI-3K or MAPK pathway. Place and Duration of Study: Department of Biological Sciences, California State University, Los Angeles, CA, USA, between July 2021 and August 2022. Methodology: Rat embryos were removed by cesarean section from mother rats at 18 days’ gestation and the hippocampi of the former dissected, plated into culture dishes, and treated with the appropriate drug(s) (see Study Design above). After 4 days, neurons were harvested for Western blotting. Optical density of Western blot bands were quantified and statistically analyzed in a 2-way ANOVA, using a level of statistical significance at P < .05. Results: Under normal conditions (with N2 supplement), ashwagandha, but not withanolide A, increased phospho-TrkB immunoreactivity when compared to the effects of vehicle (controls, F(11, 24) = 22.48, P < .001), although withanolide A did not quite reach statistical significance (P = .069) when compared to that of the controlled condition. Likewise, under nutrient-deprived conditions, both ashwagandha and withanolide A also increased phosphorylation of TrkB when compared to that of vehicle-nutrient-deprived conditions (P < .0001). The same results were obtained in the presence of inhibitors of TrkB itself and the PI-3K (ashwagandha, P < .001; withanolide A, P < .001) and MAPK (ashwagandha, P = .027; withanolide A, P = .045) pathways. Conclusion: Ashwagandha or withanolide A activates TrkB, in nutrient-deprived hippocampal neurons, underscoring its role in neuronal survival signaling.

Advances in the study of molecular identification technology of Echinococcus species

@#The larvae of Echinococcus (hydatidcyst) can parasitize humans and animals, causing a serious zoonotic disease-echinococcosis. The life history of Echinococcus is complicated, and as the disease progresses slowly after infection, early diagnosis is difficult to establish. Due to the limitations of imaging and immunological diagnosis in this respect, domestic and foreign scholars have established a variety of molecular detection techniques for the pathogen Echinococcus over recent years, mainly including nested polymerase chain reaction (PCR), multiplex PCR, real-time quantitative PCR, and nucleic acid isothermal amplification technology. In this article, the research progress of molecular detection technology for Echinococcus infection currently was reviewed and the significance of these methods in the detection and diagnosis of hydatid and hydatid diseases was also discussed.

Dendritic cell TLR4 induces Th1-type immune response against Cryptosporidium parvum infection

@#The objective of this study was to investigate the mechanism of Toll-like receptor (TLR4)- mediated dendritic cell (DC) immune against Cryptosporidium parvum infection. C. parvum sporozoites were labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester. Murine bone marrow-derived DCs were isolated, and divided into TLR4 antibody blocking (TAB; infected with 2 × 105 labeled sporozoites and 0.5 μg TLR4 blocking antibody), TLR4 antibody unblocking (TAU; infected with 2 × 105 labeled sporozoites), and blank control (BC; with 1.5 mL Roswell Park Memorial Institute 1640 medium) groups. The adhesion of Cryptosporidium sporozoites to DCs and CD11c+ levels were examined by fluorescence microscopy and flow cytometry. Male KM mice were orally injected with C. parvum. The proliferation of T lymphocytes in spleen, expression of cytokines in peripheral blood, and TLR4 distribution features in different organs were further determined by immunohistochemistry. A significantly higher expression of CD11c+ and higher C. parvum sporozoite adhesion were found in the TAU group compared with other groups. The expression of CD4+CD8- /CD8+CD4- in the spleen were obviously differences between the TAB and TAU groups. The expression of TLR4, interleukin IL-4, IL-12, IL-18 and IFN-γ improved in the TAU group compared with TAB group. Higher expression of TLR4 was detected in the lymph nodes of mice in the TAU group, with pathological changes in the small intestine. Hence, TLR4 could mediate DCs to recognize C. parvum, inducing Th1 immune reaction to control C. parvum infection.

Parasitological and molecular detection of human fascioliasis in a young man from Guizhou, China

@#A 24-year-old man born in Guizhou province was diagnosed with obstructive jaundice and bile duct stones in 2013. Four living trematodes were found during laparotomy and cholecystectomy. Based on the morphology and molecular genetics analysis of internal transcribed spacer and pcox1 genes of the flatworm specimens, the trematodes from the patient were confirmed to be Fasciola hepatica. This report provided the clinical and molecular diagnosis information on human fascioliasis, which is an emerging sanitary problem still ignored in China. Human fascioliasis constantly occurs due to climatic changes and frequency of human travel. Therefore, it deserves more attention from physicians working in both developing and developed countries.

microRNAs expression profiles in Schistosoma japonicum of different sex 14 and 28 days post-infection

@#Different miRNAs are involved in the life cycles of Schistosoma japonicum. The aim of this study was to examine the expression profile of miRNAs in individual S. japonicum of different sex before and after pairing (18 and 24 dpi). The majority of differential expressed miRNAs were highly abundant at 14 dpi, except for sja-miR-125b and sja-miR-3505, in both male and female. Moreover, it was estimated that sja-miR-125b and sja-miR-3505 might be related to laying eggs. sja-miR-2a-5p and sja-miR-3484-5p were expressed at 14 dpi in males and were significantly clustered in DNA topoisomerase III, Rap guanine nucleotide exchange factor 1 and L-serine/L-threonine ammonia-lyase. Target genes of sja-miR-2d-5p, sja-miR-31- 5p and sja-miR-125a, which were expressed at 14 dpi in males but particularly females, were clustered in kelch-like protein 12, fructose-bisphosphate aldolase, class I, and heat shock protein 90 kDa beta. Predicted target genes of sja-miR-3483-3p (expressed at 28 dpi in females but not in males) were clustered in 26S proteasome regulatory subunit N1, ATPdependent RNA helicase DDX17. Predicted target genes of sja-miR-219-5p, which were differentially expressed at 28 dpi in females but particularly males, were clustered in DNA excision repair protein ERCC-6, protein phosphatase 1D, and ATPase family AAA domaincontaining protein 3A/B. Moreover, at 28 dpi, eight miRNAs were significantly up-regulated in females compared to males. The predicted target genes of these miRNAs were significantly clustered in heat shock protein 90 kDa beta, 26S proteasome regulatory subunit N1, and protein arginine N-methyltransferase 1. To sum up, differentially expressed miRNAs may have an essential role and provide necessary information on clarifying this trematode’s growth, development, maturation, and infection ability to mammalian hosts in its complex life cycle, and may be helpful for developing new drug targets and vaccine candidates for schistosomiasis.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

Detection of acute cerebral hemorrhage in rabbits by magnetic induction

Acute cerebral hemorrhage (ACH) is an important clinical problem that is often monitored and studied with expensive devices such as computed tomography, magnetic resonance imaging, and positron emission tomography. These devices are not readily available in economically underdeveloped regions of the world, emergency departments, and emergency zones. We have developed a less expensive tool for non-contact monitoring of ACH. The system measures the magnetic induction phase shift (MIPS) between the electromagnetic signals on two coils. ACH was induced in 6 experimental rabbits and edema was induced in 4 control rabbits by stereotactic methods, and their intracranial pressure and heart rate were monitored for 1 h. Signals were continuously monitored for up to 1 h at an exciting frequency of 10.7 MHz. Autologous blood was administered to the experimental group, and saline to the control group (1 to 3 mL) by injection of 1-mL every 5 min. The results showed a significant increase in MIPS as a function of the injection volume, but the heart rate was stable. In the experimental (ACH) group, there was a statistically significant positive correlation of the intracranial pressure and MIPS. The change of MIPS was greater in the ACH group than in the control group. This high-sensitivity system could detect a 1-mL change in blood volume. The MIPS was significantly related to the intracranial pressure. This observation suggests that the method could be valuable for detecting early warning signs in emergency medicine and critical care units.

Analysis of the clinicopathological features of hepatocellular carcinoma in elderly patients.

The peak age of onset of hepatocellular carcinoma (HCC) is continually increasing worldwide. This study aims to evaluate whether there exists any significant difference in the clinicopathological features between younger- and elderly-HCC.1082 Consecutive patients with HCC who underwent liver resection at Liver Cancer Institute, Zhongshan Hospital, Fudan University from 1995 to 1998 were studied. The patients were divided into elderly-HCC (>or=65 years of age) and younger-HCC (< 65 years of age). Important clinicopathological features of the patients and postoperative survival rates were compared between these two groups. Among 1082 patients studied, 108 were elderly-HCC and 974 were younger-HCC. The resection rate of the elderly-HCC was significantly higher than that of the younger-HCC. The 1, 3 and 5-year survival rates of the elderly-HCC were not significantly different from those of the younger-HCC. Compared with the younger-HCC, the elderly-HCC had (1) less HBsAg-positive rate; (2) more frequent anti-HCV positivity ; (3) lower proportion with AFP value >or=400 microg/dl; (4) a relatively small tumor diameter; (5) higher proportion of stage I-II patients; (6) a relatively low metastasis rate. However, there were no statistically significant differences in other clinicopathological features (including gender, symptoms, tumor number, tumor venous invasion, tumor differentiation, capsular formation, type of cirrhosis) between the two groups. There is a certain extent difference in clinicopathological features between elderly and younger-HCC patients, but the postoperative survival rate is comparable between the two groups.

Study on detecting antibodies to Toxoplasma gondii in pooled serum of blood donors by Dot-IGSS.

To prevent post-transfusion toxoplasmosis, the sera of blood donors, per six of which were mixed as a pool, were examined for anti-Toxoplasma gondii antibiodies by Dot-immunogold silver staining (Dot-IGSS) with the single serum specimens examined simultaneously. The results showed that the sensitivity and specificity of serum pool method were 92.31% and 99.96% respectively. The consistent rate between the two methods was 99.73% and kappa value was 0.947 (p<0.01). Considering the mean infection rate of Toxoplasma gondii being 4.86% in China, if the serum pool method be adoped, with pool size k=5, a 57% reduction in the number of tests, as well as the cost of the screen, can be expected. Beside the social benefit, consequent upon the interruption of the Toxoplasma gondii infection spread through blood transfusion also can be expected.

Hydroxychloroquine sulphate inhibits in vitro apoptosis of circulating lymphocytes in patients with systemic lupus erythematosus.

The serological hallmark of systemic lupus erythematosus (SLE) is the presence of antibodies against double-stranded DNA. However, several studies have suggested that it is not DNA itself, but nucleosomes that are the immunogenic particles involved both in the induction of anti-DNA antibodies, and in the pathophysiology of SLE. Meanwhile, It has been demonstrated that there is an accelerated in vitro apoptosis of lymphocytes from patients with SLE. Therefore, one can postulate that the process of apoptosis may provide a source of nuclear antigens to drive the autoantibody response seen in SLE. Our study has demonstrated that hydroxychloroquine exhibits an anti-apoptotic action and this anti-apoptotic effect is dependent on monocyte coexistence. We used both morphology assessment and fluorescent antibody cell sorter (FACS) analysis to measure the apoptotic percentage of lymphocytes from 25 SLE patients in medium alone (control) or with the addition of different concentrations of hydroxychloroquine. Our results have shown that there is a significant decrease in the percentage of apoptosis at the therapeutic concentration (10(-6) M) as compared with the control (p < 0.05). It has been reported that the anti-rheumatic properties of hydroxychloroquine result from its interference with antigen processing in macrophages and other antigen-presenting cells. We propose that this results in decreased stimulation of autoreactive lymphocytes reactive with self-peptides, and consequently diminution of activation-induced cell death (apoptosis) of mature peripheral lymphocytes.

Combinations of three immunological assays for detecting anti-Toxoplasma IgG in the sera of patients infected with Toxoplasma gondii.

Enzyme-linked immunosorbent assay (ELISA), Dot-ELISA and Dot-immunogold silver staining (Dot-IGSS) were simultaneously used to detect the specific IgG against Toxoplasma gondii in 65 patients infected with the protozoa. The positive rates were 86.51%, 92.51% and 98.64%, respectively. When ELISA and Dot-ELISA results were put together, the positive rate increased to 95.38%. When Dot-IGSS results were combined with those of ELISA or Dot-ELISA, the positive rate was raised to 100%. The difference in positive rate between ELISA and Dot-IGSS was significant (x2 = 6.93, p < 0.01), but no statistically significant differences were found between ELISA and Dot-ELISA or between Dot-ELISA and Dot-IGSS. Paired comparison of the reacting intensities of the sera in the 3 assays showed the correlations were highly significant (p < 0.001), with r = 0.608 between Dot-IGSS and Dot-ELISA, r = 0.8194 between Dot-IGSS and ELISA and r = 0.517 between Dot-ELISA and ELISA. Hence combination of different serological assays may increase their sensitivity and specificity for detecting the anti-Toxoplasma antibodies.

Comparative study of Dot-immunogold silver staining and Dot-ELISA for the detection of serum antibodies against Wuchereria bancrofti.

Dot-immunogold silver staining (Dot-IGSS) and Dot-ELISA, using the soluble antigen of Brugia malayi, were employed to detect anti-Wuchereria bancrofti antibodies in 50 cases of Wuchereria bancrofti microfilaremia. The positive rates were 100% and 90% in Dot-IGSS and Dot-ELISA respectively. The average titer in the 45 positive cases was 1:184 (1:10-1:2560) for Dot-IGSS and 1:150 (1:10-1:2560) for Dot-ELISA, with 30 cases showing the same titer in both tests, 13 cases showing higher titer in Dot-IGSS than in Dot-ELISA and 2 cases in the former showing lower titers than in the latter. There was a linear relationship between the titers of antibodies detected by Dot-IGSS and by Dot-ELISA (r = 0.8443). Dot-IGSS, similar to Dot-ELISA, is easy to carry out and the result is easy to read. It is seen that Dot-IGSS is highly sensitive and specific and is practicable for immunodiagnosis and surveillance of filariasis.

Monoclonal gammopathies and the related autoimmune manifestations in Taiwan.

A total of 50,000 patients were surveyed for the presence of monoclonal immunoglobulins during the past two decades. There were 411 cases of monoclonal gammopathies including 243 cases of plasma cell neoplasms and 168 cases of secondary plasma-cell dyscrasia. Among the 227 cases of multiple myeloma and Waldenström's macroglobulinemia, there were 49.3% IgG class, 22.9% IgA class, 9.7% IgM class and 13.2% light chain type. In addition, there were 1.3% of nonexcretory myeloma including an IgM type. A relatively high frequency (4.8%) of IgD M-proteins was detected but heavy chain disease was not encountered in the present series. Purified M-components from patients with possible autoimmune manifestations were subjected to immunofluorescence studies. Autoimmune activity of M-proteins was found in a patient of Waldenström's macroglobulinemia with peripheral neuropathy, and another patient of cryofibrinogenemia with recurrent purpura and gangrene. In conclusion, a high frequency of IgD myeloma is found in Chinese patients of this area. M-components may have autoimmune activity resulting in unusual clinical manifestations.

Status and trend of HIV-1 infection and AIDS in Taiwan, December, 1991.

From May 1, 1985 to December 31, 1991, a total of 4,962,707 serum samples from 8 population groups in Taiwan were tested for anti-human immunodeficiency virus type 1 (anti-HIV-1). In total, 256 samples were seropositive; of these individuals, 43 developed acquired immunodeficiency syndrome (AIDS): 29 were homosexuals; 5 were hemophiliacs; 8 were heterosexuals and 1 was of unknown risk. Although the prevalence of HIV-1 infection and AIDS remains low compared with other countries, since 1988 the increase has been rapid. Before 1977 the majority were homosexuals and hemophiliacs; thereafter the risk groups diversified, with a trend away from homosexuals and hemophiliacs towards heterosexuals and intravenous drug abusers (IVDAs). A few patients have caused serious social problems for the public, health care workers and families. Active community efforts are needed to achieve future success in the control of HIV-1 infection and AIDS in Taiwan.

Fasciola hepatica infection in China.

Fasciola hepatica is one of the most common parasites in China that infects ruminants, especially cattle, sheep and goats, and causes considerable morbidity and economic losses in the animal husbandry. The climate in wide areas of China is suitable for the development of F. hepatica and its intermediate snail hosts. However, human infection with the fluke is not common. Up to now, 38 hospital patients and six infected individuals from two epidemiological surveys have been reported in the Chinese literature. F. gigantica infection in herbivorous animals is comparatively rare compared to F. hepatica infections, and only one case was reported in man (Wang, 1984). This paper reviews mainly human infections with F. hepatica in the Chinese literature. Animal infections of the parasite are also briefly described.

Dose finding double-blind clinical trial with praziquantel in schistosomiasis japonica patients.

A double-blind clinical trial with praziquantel was carried out. A total of 400 cases was treated with four different dosages, namely, 60, 50, 40 and 30 mg/kg body weight of praziquantel. The drug was given in one day divided into two doses. Identical placebo tablets were used to make up a total of 60 mg/kg. Tolerance was good in all with the exception of one case suffering from asthmatic attack with papule rashes over large area of the body surface. 394 patients were able to be followed up parasitologically six months post-treatment. 79.8%, 71.7%, 78.8% and 70.1% of the patients were negative in the groups with the total dose of 60, 50, 40 and 30 mg/kg respectively. The cure rates as well as the side effects were similar for the four groups. The efficacy was lower than that reported by other authors and the possibility of reinfection was incriminated. In villages with few snails the negative hatching rates in aforementioned four groups were 89.1%, 91.1%, 88.9% and 81.8%, while in villages with abundant snails the rates were 68.2%, 46.5%, 66.7% and 54.8%. The difference between the two areas was statistically significant. Higher efficacy was observed in adults with an average cure rate of 80.0% than in children under 15 years of age, the average cure rate being 57.1%.(ABSTRACT TRUNCATED AT 250 WORDS)