Blinding assessment in clinical trials of traditional Chinese medicine: Exploratory principles and protocol.

As one of the key components of clinical trials, blinding, if successfully implemented, can help to mitigate the risks of implementation bias and measurement bias, consequently improving the validity and reliability of the trial results. However, successful blinding in clinical trials of traditional Chinese medicine (TCM) is hard to achieve, and the evaluation of blinding success through blinding assessment lacks established guidelines. Taking into account the challenges associated with blinding in the TCM field, here we present a framework for assessing blinding. Further, this study proposes a blinding assessment protocol for TCM clinical trials, building upon the framework and the existing methods. An assessment report checklist and an approach for evaluating the assessment results are presented based on the proposed protocol. It is anticipated that these improvements to blinding assessment will generate greater awareness among researchers, facilitate the standardization of blinding, and augment the blinding effectiveness. The use of this blinding assessment may further advance the quality and precision of TCM clinical trials and improve the accuracy of the trial results. The blinding assessment protocol will undergo continued optimization and refinement, drawing upon expert consensus and experience derived from clinical trials. Please cite this article as: Wang XC, Liu XY, Shi KL, Meng QG, Yu YF, Wang SY, Wang J, Qu C, Lei C, Yu XP. Blinding assessment in clinical trials of traditional Chinese medicine: Exploratory principles and protocol. J Integr Med. 2023; 21(6): 528-536.

[Priority ranking of Chinese patent medicine prescriptions for type 2 diabetes mellitus (syndrome of deficiency of both Qi and Yin): based on analytic hierarchy process (AHP)].

Clinical expertise, patient preference, and the best evidence are the three elements of evidence-based medicine. Based on high-level and high-quality evidence, qualitative and quantitative analysis of the prescribing decisions of physicians is beneficial to improving clinical efficacy. A mature methodological system is available for the retrieval, analysis, summary, evaluation, and recommendation of the evidence, but there are still few studies on physicians' prescribing decisions. How to analyze the trend of physicians' prescribing decisions based on the priority ranking in addition and subtraction of prescriptions? Analytic hierarchy process(AHP) is a method for decision making, which arranges the elements of the decision problem into overall goal, criteria, and operational sub-criteria, and uses the matrix eigenvector method to solve the problem. This study aims to analyze the priority of physicians' prescribing decisions for diabetes mellitus with deficiency of both Qi and Yin based on AHP. To be specific, a database of diabetes mellitus cases with deficiency of both Qi and Yin was established and AHP was used to yield the priority ranking of Chinese patent medicine prescriptions in specific clinical scenarios. In the selected cases of diabetes mellitus with deficiency of both Qi and Yin, Xiaoke Pills was the best prescription for the treatment of type 2 diabetes mellitus(deficiency of both Qi and Yin)(normalized=0.388), followed by Liuwei Dihuang Pills(normalized=0.269), Qishen Capsules(normalized=0.230), and Shengmai Injection(normalized=0.113). According to the analysis the available data, for type 2 diabetes mellitus(deficiency of both Qi and Yin), Xiaoke Pills was the most effective prescription in specific scenarios. When the physicians' prescribing decisions are consistent with the evidence, quantitative analysis of physicians' cognition will boost the evidence-based medical decision-making. However, the research results are also affected by the quality of literature, evidence level and priority, which are thus have some limitations. It is recommended that further small data research based on individual cases be carried out to lay a evidence-based basis for the clinical decision-making of type 2 diabetes mellitus.

A metabolomic approach to elucidate the inhibitory effects of baicalin in pulmonary fibrosis.

CONTEXT: Baicalin, a major flavonoid extracted from Scutellaria baicalensis Georgi (Lamiaceae), has been shown to exert therapeutic effects on pulmonary fibrosis (PF). OBJECTIVE: To use serum metabolomics combined with biochemical and histopathological analyses to clarify anti-PF mechanisms of baicalin on metabolic pathways and the levels of potential biomarkers. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were randomly divided into the control, PF model, prednisolone acetate-treated (4.2 mg/kg/day) and baicalin-treated (25 and 100 mg/kg/day) groups. A rat model of PF was established using a tracheal injection of bleomycin, and the respective drugs were administered intragastrically for 4 weeks. Histomorphology of lung tissue was examined after H&E and Masson's trichrome staining. Biochemical indicators including SOD, MDA and HYP were measured. Serum-metabonomic analysis based on UPLC-Q-TOF/MS was used to clarify the changes in potential biomarkers among different groups of PF rats. RESULTS: Both doses of baicalin effectively alleviated bleomycin-induced pathological changes, and increased the levels of SOD (from 69.48 to 99.50 and 112.30, respectively), reduced the levels of MDA (from 10.91 to 5.0 and 7.53, respectively) and HYP (from 0.63 to 0.41 and 0.49, respectively). Forty-eight potential biomarkers associated with PF were identified. Meanwhile, the metabolic profiles and fluctuating metabolite levels were normalized or partially reversed after baicalin treatment. Furthermore, baicalin was found to improve PF potentially by the regulation of four key biomarkers involving taurine and hypotaurine metabolism, glutathione metabolism, and glycerophospholipid metabolism. CONCLUSIONS: These findings revealed the anti-fibrotic mechanisms of baicalin and it may be considered as an effective therapy for PF.

Retracted: The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro.

The authors repeated experiments and found that the results shown in figure 2 were not reproducible. Reference: Shuang-li Zhang, Bao-lin Li, Wei Li, Ming Lu, Lin-ying Ni, Hui-li Ma, Qing-gang Meng. The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro. Med Sci Monit 2018; 24: LBR4926-4933. 10.12659/MSM.909193.

Extracorporeal shockwave therapy for the treatment of knee osteoarthritis: A retrospective study.

This retrospective study investigated the effect and safety of extracorporeal shockwave therapy (ESWT) for treatment of knee osteoarthritis (KOA).In this retrospective study, 105 patients with KOA were included. Of those, 60 patients underwent ESWT, whereas 45 patients received laser therapy. Effect was measured by the Numeric Rating Scale (NRS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). In addition, adverse events (AEs) were also recorded in this study. All outcome measurements were evaluated at the end of weeks 1, 6, and 12.Compared with the laser therapy, ESWT showed greater effect in KOA symptoms relief with regard to NRS, WOMAC total, and its subscores at week 6 (P < .05) and week 12 (P < .01) after treatment. No AE, however, occurred in both groups.The results of this retrospective study found that ESWT may be efficacious and safe for the treatment of patients with KOA. It, however, had an intrinsic limitation as a retrospective study. Prospective study with larger sample size is still needed to warrant the result of this study in the future.

The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro.

BACKGROUND The aim of this study was investigate the effects of the sesquiterpene lactone, ludartin, on cell proliferation, cell migration, apoptosis, and the cell cycle in osteosarcoma cell lines, compared with a normal osteoblast cell line. MATERIAL AND METHODS Osteosarcoma cell lines, MG-63 Saos-2 U-2OS, T1-73 143B, and HOS, and normal hFOB 1.19 osteoblasts, were cultured and treated with increasing doses of ludartin, The MTT colorimetric assay was used to measure cell metabolic activity and viability. Apoptosis was studied by fluorescence-activated cell sorting (FACS) using 4',6-diamidino-2-phenylindole (DAPI) nuclear staining and Annexin-V/propidium iodide (PI) staining. Cell cycle was studied using flow cytometry. Cell migration and invasion were studied using wound healing and Boyden chamber assays. Protein expression was measured by Western blotting. RESULTS Ludartin inhibited cell viability, cell migration, cell proliferation, and increased cell apoptosis, in all osteosarcoma cell lines, with an IC50 dose ranging from 15-30 µM. The greatest effects were on the Saso-2 osteosarcoma cells, with an IC50 of 15 µM. However, ludartin showed minor cytotoxic effects of the normal hFOB 1.19 osteoblasts (IC50 >100 µM). Ludartin exerted its anti-proliferative effects on Saos-2 cells via induction of apoptosis and cell cycle arrest at the G2/M checkpoint, associated with reduced expression of Cdc25c (Ser216), Cdc25c, pCdc2 (Tyr15), and Cdc2 and increased expression of p21WAF1. Ludartin inhibited cell migration and invasion of the Saos-2 cells. CONCLUSIONS The dose-dependent effects of ludartin on cell proliferation, migration, apoptosis, cell cycle arrest at the G2/M checkpoint involved p21WAFI in Saos-2 osteosarcoma cells.

Effect of extracorporeal shock-wave therapy for treating patients with chronic rotator cuff tendonitis.

BACKGROUND: This study aimed to determine the efficacy and safety of extracorporeal shock-wave therapy (ESWT) for treating patients with chronic rotator cuff tendonitis (CRCT). METHODS: In this study, 84 patients with CRCT were randomly divided into intervention and control groups in a ratio of 1:1. Patients in the intervention group received ESWT, whereas those in the control group received placebo. The primary outcome was measured by Numeric Rating Scale (NRS). The secondary outcomes were measured by the Constant-Murley score (CMS), simple shoulder test (SST) score, and adverse events. RESULTS: Compared with placebo, ESWT showed greater efficacy in shoulder pain relief with regard to NRS score and shoulder function as measured by using CMS and SST score at 4 weeks (P < .05) and 8 weeks (P < .01) after treatment. However, no adverse events occurred in both groups. CONCLUSION: ESWT was efficacious and safe for treating patients with CRCT.

Effect and safety of sertraline for treat posttraumatic stress disorder: a multicenter randomised controlled study.

OBJECTIVE: Although several previous studies have examined the efficacy of sertraline in the treatment of posttraumatic stress disorder (PTSD), none involved Chinese PTSD patients. This study aimed to evaluate sertraline efficacy and adverse events in Chinese patients with PTSD over 12 weeks. METHODS: In total, 72 PTSD patients were randomly assigned to receive sertraline (135 mg daily) or a placebo for 12 weeks. Impact of Event Scale-Revised subscores constituted the primary outcome, with Clinical Global Impression Scale-Severity scores and adverse events as secondary outcomes. RESULTS: Sixty-five subjects completed the study, and their data were included in the final analysis. Sertraline showed greater efficacy in enhancing Impact of Event Scale-Revised and Clinical Global Impression Scale-Severity scores at 6 and 12 weeks relative to that of the placebo. The most common adverse event was nausea, which occurred in 12 (33.3%) and 8 (22.2%) patients in the sertraline and placebo groups, respectively. No sertraline-related deaths were recorded. CONCLUSIONS: In summary, we demonstrated that 12 weeks of sertraline was efficacious and well-tolerated in Chinese patients with PTSD.

Predicting factors associated with frailty in aged patients with bone-arthrosis pain in the clinic.

OBJECTIVES: To identify frail and pre-frail patients in a group of patients older than 60 years. METHODS: The phenotype model of Fried's method was used to identify frailty and pre-frailty in total of 78 participants. Cognitive ability and psychosocial function tests were also given to 59 of the 78 patients. RESULTS: Prevalence of frailty and pre-frailty was 14.1% (11/78) and 46.2% (36/78), respectively. Of the 5 phenotype variables, weak grip strength was the most commonly seen variable with 53.8% of all participants and 100% in the frail group. Low energy expenditure, however, was not self-reported by any participant in the current study (0%). Prevalence of frailty in the present study is associated with chronological age. The current study indicates that 4 phenotypic variables (unintentional weight loss, self-reported exhaustion, gait speed and grip strength) contribute to the development to frailty, and that cognitive impairment and psychosocial frailty also predict frailty or pre-frailty in the patients older than 60 years old irrespective of chronic pain or osteoarthritis. The findings of the current study suggest frailty and pre-frailty are common in senior Chinese patients with chronic diseases. CONCLUSION: Recognition and identification of frailty in a rehabilitation clinic or hospital might help physicians to provide appropriate counseling to patients and families about adverse outcomes of certain treatments such as surgery, and could optimize management of coexisting chronic diseases that might contribute to or be affected by frailty.

Acupuncture for the Treatment of Oculomotor Paralysis: A Pilot Randomised Controlled Trial.

This study consisted of a single centre randomised controlled trial with two parallel arms: an acupuncture group (n = 20) with 27 affected eyes and a sham group (n = 20) with 23 affected eyes. Participants in the acupuncture group received acupuncture treatment once daily, three times weekly for four weeks. Participants assigned to the control group received sham acupuncture, the same protocol as that used for the acupuncture group but without insertion of needles into the skin. The primary outcome measure was the cervical range of motion (CROM) score. Secondary outcome measures were the palpebral fissure size, response rate, and adverse events. All 40 participants completed the study. In the comparison of acupuncture and sham acupuncture, a significant difference was observed between acupuncture and sham acupuncture in CROM score (21.37 ± 15.16 and 32.21 ± 19.54, resp.) (P < 0.05) and palpebral fissure size (7.19 ± 2.94 and 5.41 ± 2.45, resp.) (P < 0.05). Response rate was also significantly different in the acupuncture group (P < 0.05). No adverse events were reported in both groups in this study. In summary, it was demonstrated that acupuncture had a feasibility positive effect on oculomotor paralysis.

Effect of long-term pulsed electromagnetic field exposure on hepatic and immunologic functions of rats.

BACKGROUND: In this report, the effects of long-term pulsed electromagnetic field (PEMF) exposure on hepatic and immunologic functions were examined. METHODS: Male rats were randomly divided into four groups: a control group and three experimental groups exposed to a 50-Hz PEMF at 5, 10, or 20 mT for 10 weeks. RESULTS: Compared with the control group, activities of serum alanine aminotransferase and aspartate aminotransferase and concentrations of serum, liver, and spleen Metabolism of lipid peroxidation (MDA) in the 10- and 20-mT PEMF groups were significantly increased. The activities of Glutathione peroxidase (GSH-Px) and Superoxide Dismutase (SOD) in the serum, liver, and spleen and concentrations of serum immunoglobulins were significantly decreased. CONCLUSION: These results demonstrate that long-term exposure to PEMF can lead to oxidative damage of the liver and spleen.

Apoptosis induced by sonodynamic therapy in human osteosarcoma cells in vitro.

The aim of the present study was to investigate the potential effect of hematoporphyrin monomethyl ether-sonodynamic therapy (HMME-SDT) on MG-63 osteosarcoma cells. The HMME concentration was kept constant at 20 µg/ml and the MG-63 osteosarcoma cell line was exposed to ultrasound with an intensity of 1.0 W/cm2 for 30 sec. Cell cytotoxicity was quantified using an MTT assay 6 h after HMME-SDT. The intracellular localization of HMME was imaged using inverted confocal laser scanning microscopy. Apoptosis was investigated using flow cytometry with Annexin V-fluorescein isothiocyanate and propidium iodine staining. The cytotoxicity of HMME-mediated sonodynamic action on MG-63 cells was significantly higher than that of other treatments, including ultrasound alone, HMME alone and sham treatment. Flow cytometry demonstrated that HMME­SDT action markedly enhanced the apoptotic rate of MG-63 cells. The mechanisms of apoptosis were analyzed by measuring the protein expression of poly ADP-ribose polymerase (PARP), cleaved PARP, procaspase-3, cleaved caspase-3 and cleaved caspase-9. The data demonstrated that HMME-SDT action markedly induced the apoptosis of MG-63 cells.

[Relationship between effect of sacral canal injection and different sections, types, courses of lumbar intervertebral disc herniation].

OBJECTIVE: To observe the effects of sacral canal injection in treating lumbar intervertebral disc herniation with different sections, types and courses, in order to provide selective basis for clinical indications. METHODS: From March 2007 to May 2011, 329 patients with lumbar intervertebral disc herniation were treated with sacral canal injection and their data were retrospectively analyzed. There were 178 males and 151 females,ranging in age from 16 to 78 years old with an average of 45 years; ranging in course of disease from 2 days to 41 years with an average of 4.1 years. All the protrusions were simple segment, and L3,4 of 51 cases, L4,5 of 142 cases, L5S1 of 136 cases including central protrusion of 137 cases, posterolateral of 145 cases, extreme posterolateral of 47 cases. According to different sections, types, courses (more than or less than 6 months ) of lumbar intervertebral disc herniation, the patients were divided into several groups, clinical effects of the groups were compared and analyzed. RESULTS: Clinical effect of protrusion in L5S1 group was better than that of L3,4, L4,5 group; the effect of patients with course less than 6 months was better than with course more than 6 months; effect of posterolateral protrution was better than that of central and extreme posterolateral protrusion. Clinical effect of corresponding the three conditions was better than that of others no corresponding the three conditions,the three conditions included the course less than 6 months, protrusion in L5S1 section, with type of posterolateral protrusion. No complications were found in all patients and cured patients were followed up for 1 year with the recurrence rate of 0.05%. CONCLUSION: The patients with short course, lower protrusion, type of posterolateral protrusion can obtained better effects, while the patients who meets the three conditions is appropriate mostly to sacral canal injection therapy.

Decreased expression of CD9 in bovine oocytes after cryopreservation and the relationship to fertilization capacity.

This study was conducted to investigate the effect of vitrification of bovine metaphase-II (MII) oocytes on CD9 expression and fertilization capacity. Surviving vitrified/warmed oocytes were used to detect CD9 distribution (fluorescence microscopy), CD9 mRNA (qRT-PCR), and CD9 protein expression (Western blot), and to analyze in vitro fertilization rates (number of sperm bound to or that penetrated the oocytes) after removing the zona pellucida. Fresh oocytes acted as control. The experimental results showed that the vitrification/warming procedures significantly decreased CD9 expression at the mRNA and protein levels, and changed the CD9 distribution pattern in bovine oocytes. After fertilization in vitro, the average number of sperm binding and penetration of vitrified oocytes were significantly lower than those of the non-vitrified oocytes. In conclusion, vitrification of bovine oocytes caused a decrease in CD9 expression at the mRNA and protein levels, and an alteration of CD9 distribution pattern, which may have resulted in lowered fertilization capacity.

Revision hip arthroplasties with use of the modular S-ROM prosthesis.

OBJECTIVE: The aim of this study was to revision hip arthroplasties in ankylosing spondylitis. MATERIALS AND METHODS: We performed 21 revision hip arthroplasty in 16 patients with ankylosing spondylitis (AS) using S-ROM modular femoral stem. At the final follow-up, all patients were able to ambulate without any walking aids. RESULTS: The mean Harris hip score improved from 57.2 (35 to 74) pre-operatively to 90.2 points (73 to 100) post-operatively, and the outcome was classified as good or excellent in 17 hips (81%). Fixation of the femoral component was classified as stable with bone ingrowth in 15 hips (71.4%), stable with fibrous ingrowth in 2 hips (9.5%), and radiolucent loosening in 4 hips (19.1%). Five hips developed a pedestal at the tip of femoral component. Femoral osteolysis was found in 3 hips (14.2%): 3 hips in Gruen zones 1 and 7, one hip in zone 7, and two hips in zone 1. One hip underwent acetabular revision because of breakage of polyethylene liner, and the well-fixed femoral component was left in situ. Thigh pain developed in 1 patient (6.2%). Kaplan-Meier survival was 81% at 64 months, with radiographic loosening as an end-point when two hips were at risk. CONCLUSION: Satisfactory results of midterm clinical and radiographic follow-up can be achieved using S-ROM modular femoral stem for revision of femoral stem in ankylosing spondylitis.

In vitro derivation of germ cells from embryonic stem cells in mammals.

Previous reports have shown that embryonic stem (ES) cells, derived from the inner cell mass of mouse or human blastocysts, could differentiate in vitro into female and male germ cells as well as into the cell types of all three germ layers. While in one case, the ES cell-derived germ cells have been reported to give birth to live offspring in the mouse, these cells differ in fertilization capacity from the sperm and oocytes produced in vivo as they cannot complete meiosis under in vitro conditions. The efficiency of functional germ cell isolation from ES cells is also low. According to published reports, factors such as the proper selection of feeder cells, including ovarian granulosa cells and those which could secrete bone morphogenic protein-4 (BMP4), and the addition of retinoic acid into culture medium, could to some extent establish and improve the microenvironment ES cells rely on for differentiation into germ cells. This review briefly describes the progress of deriving germ cells from ES cells and discusses possible factors that could improve in vitro gamete production.

Changes in acetylation on lysine 12 of histone H4 (acH4K12) of murine oocytes during maternal aging may affect fertilization and subsequent embryo development.

OBJECTIVE: To compare acH4K12 levels in oocytes during mouse aging and then assess how such changes might affect the developmental potential of oocytes. DESIGN: Experimental animal study. SETTING: State key laboratory and university research laboratory. ANIMAL(S): Kunming white strain mice. INTERVENTION(S): Oocytes obtained from TSA treated group or aging mouse group were fertilized and the formation of pronuclei and subsequently developmental potential in vitro or in vivo were assessed. MAIN OUTCOME MEASURE(S): AcH4K12 levels in oocytes were assessed using fluorescence staining, and confocal microscopy and oocyte developmental potentials were determined by in vitro or in vivo methods. RESULT(S): The AcH4K12 levels in oocytes statistically significantly increased during mouse aging. When histone acetylation of oocytes of young mice was artificially increased by trichostatin A (TSA) treatment, the acH4K12 levels in male and female pronuclei in fertilized oocytes showed statistically significant changes. About 38.9% of TSA-treated oocytes failed to form pronuclei or formed morphologically abnormal pronuclei 6 hours after fertilization, which statistically significantly decreased the blastocyst rate of TSA-treated oocytes when compared with the control group (41.5% vs. 60.5%). A similar reduction in blastocyst development was also observed when oocytes collected in older mice were compared with younger mice (17.3% vs. 69.4%). CONCLUSION(S): The AcH4K12 levels in oocytes statistically significantly increased during the aging process in mice, and such changes may affect the acetylation patterns and morphology of pronuclei during fertilization and lead to a reduction in oocyte developmental potential.

[A multi-centered randomized double-blinded controlled clinical study on efficacy of gulling pa'an capsule in treating Parkinson's disease].

OBJECTIVE: To objectively evaluate the clinical efficacy of Gulling Pa'an Capsule (GPC), a Chinese medicine, in treating Parkinson's disease (PD). METHODS: According to the good clinical practice (GCP) principle, a multi-centered, double-blinded, layered, randomized and grouping-controlled clinical trial was carried out from May 2002 to January 2005 on 242 PD patients. Among them, 53 patients who had never received levodopa were randomized into two groups, 28 in group A treated with GPC, and 25 in group B treated with placebo; patients who had received levodopa were assigned depending on the Hoehn & Yahr (H-Y) grade, to 4 groups, 75 and 19 of grade 1.5 -3 in group C and E, respectively, 79 and 16 of grade 4 in group D and F, respectively, patients in group C and E were treated with GPC and Levodopa, and those in group D and F treated with placebo and Levodopa for control. The treatment course was 12 weeks for all. Changes of unified Parkinson's disease rating scale (UPDRS) II/III scores in comparing with the baseline were assessed. For the groups C, D, E and F, the dosage of levodopa administered was also recorded. Meanwhile, the blood pressure, pulse rate, blood and urine routine, liver and renal functions, electrocardiogram (ECG) and adverse reactions were monitored as the indices for safety supervise. RESULTS: (1) After treatment, symptoms were markedly improved in 1 out of the 28 patients in group A and improved in 11, the markedly improving rate was 3.6% and the improving rate 39.3%; while in group B, the corresponding outcomes were 0 (0/25) and 28.0% (7/25) respectively, showing insignificant difference between the two groups. UPDRS scores, including the total, II and III scores were all significantly lowered in group A after treatment (P < 0.01, P < 0.05); while in group B, significant lowering only showed in terms of UPDRS III (P < 0.05); but the inter-group comparison of the changes in all the three items showed no significant difference. (2) The significant improving rate was 12.0% (9/75) and improving rate 48.0% (36/75) in group C, while those in group D, 12.7% (10/79) and 24.1% (19/79) respectively, the efficacy in group C was better (P < 0.05). The items of 3 UPDRS scores in groups C and D were all significantly lowered after treatment (P < 0.01), and the lowering in group C was more significant in terms of the total and II scores (P < 0.05). (3) The significant improving rate was 5.3% (1/19) and improving rate 36.8% (7/19) in group E, while in group F 0% (0/19) and 25.0% (4/16), respectively, showing insignificant difference between them; UPDRS scores lowered significantly in the two groups after treatment (P < 0.01), also showed no statistical significance in comparison (P > 0.05). (4) The dosage of Levodopa required in groups C and E was significantly reduced after treatment (P < 0.05), while in groups D and F, it was unchanged (P > 0.05); yet, the further analysis displayed that significant reduction only presented in group C (P < 0.05), not in the other three groups. CONCLUSIONS: The overall efficacy of levodopa in combined with GPC for treating PD patients of H-Y grade 1.5 -3 is significantly higher than that of levodopa alone. GPC shows obvious effects in improving patients' motor syndrome and the quality of life; as used in combining with levodopa, the dosage of levodopa required could be reduced.

Tetraspanin CD9 in bovine oocytes and its role in fertilization.

This study was conducted in bovine to investigate whether CD9 (a member of the tetraspanin superfamily of proteins) is present on oocytes and whether it functions in sperm-oocyte binding and fusion. First, the presence of CD9 in bovine matured oocytes was examined by immunofluorescence with the anti-CD9 monoclonal antibody (mAb) and fluorescein isothiocyanate-conjugated goat anti-mouse antibody, and the results showed that CD9 was expressed on the plasma membrane of matured oocytes. Sperm binding and fusion with oocytes was then examined by in vitro fertilization. When the zona pellucida-free matured oocytes were fertilized, both sperm binding to ooplasma and sperm penetrating into oocytes were significantly (P<0.01) reduced in anti-CD9 antibody-treated oocytes (6.3 +/- 0.7 per oocyte and 41.6%, respectively) compared with untreated control oocytes (19.0 +/- 0.7 per oocyte and 81.3%, respectively), indicating that the anti-CD9 mAb potentially inhibits sperm-oocyte binding and fusion. These results demonstrated that the CD9 present on bovine matured oocytes is involved in sperm-oocyte interaction during fertilization.

OPS vitrification of mouse immature oocytes before or after meiosis: the effect on cumulus cells maintenance and subsequent development.

Cryopreservation can cause cumulus cell damage around the immature oocytes, which may result in poor subsequent development. To evaluate the effect of the meiosis stage on the cumulus cell cryoinjury and determine the suitable stage for cryopreservation in immature oocytes, mouse oocytes at germinal vesicle (GV) and germinal vesicle breakdown (GVBD) stages were vitrified using open pulled straw (OPS) method. Cumulus cells damage was scored immediately after thawing by double-fluorescent staining. The survival rate of the oocytes was evaluated and the subsequent development of oocytes was assessed through in vitro culture (IVC) and in vitro fertilization (IVF) separately. After vitrification, a higher proportion of cumulus cells of GV oocytes were damaged than those of GVBD and untreated control groups. The survival rate of vitrified GVBD oocytes (94.1%) was significantly higher (p < 0.05) than that of GV oocytes (85.4%). Oocytes vitrified at GVBD stage (55.7%) showed similar cleavage rate compared to those at GV stage (49.2%), but significantly higher (p < 0.05) blastocyst rate (40.9% vs. 27.4%). These results demonstrate that oocytes at GVBD stage remain better cumulus membrane integrity and developmental ability during vitrification than those at GV stage, indicating they are more suitable for immature oocytes cryopreservation in mice.