Hepatic and renal effects of oral stingless bee honey in a streptozotocin-induced diabetic rat model

BACKGROUND Diabetes is known damage the liver and kidney,leading to hepatic dysfunction and kidney failure.Honey is believed to help in lowering the blood glucose levels of diabetic patients and reducing diabetic complications.However,the effect of stingless bee honey(SBH)administration in relieving liver and kidney damage in diabetes has not been well-studied.AIM To investigate the effect of SBH administration on the kidney and liver of streptozotocin-induced(STZ;55 mg/kg)diabetic Sprague Dawley rats.METHODS The rats were grouped as follows(n=6 per group):non-diabetic(ND),untreated diabetic(UNT),metformin-treated(MET),and SBH+metformin-treated(SBME)groups.After successful diabetic induction,ND and UNT rats were given normal saline,whereas the treatment groups received SBH(2.0 g/kg and/or metformin(250 mg/kg)for 12 d.Serum biochemical parameters and histological changes using hematoxylin and eosin(H&E)and periodic acid–Schiff(PAS)staining were evaluated.RESULTS On H&E and PAS staining,the ND group showed normal architecture and cellularity of Bowman’s capsule and tubules,whereas the UNT and MET groups had an increased glomerular cellularity and thickened basement membrane.The SBH-treated group showed a decrease in hydropic changes and mild cellularity of the glomerulus vs the ND group based on H&E staining,but the two were similar on PAS staining.Likewise,the SBME-treated group had an increase in cellularity of the glomerulus on H&E staining,but it was comparable to the SBH and ND groups on PAS staining.UNT diabetic rats had tubular hydropic tubules,which were smaller than other groups.Reduced fatty vacuole formation and dilated blood sinusoids in liver tissue were seen in the SBH group.Conversely,the UNT group had high glucose levels,which subsequently increased MDA levels,ultimately leading to liver damage.SBH treatment reduced this damage,as evidenced by having the lowest fasting glucose,serum alanine transaminase,aspartate transaminase,and alkaline phosphatase levels compared to other groups,although the levels of liver enzymes were not statistically significant.CONCLUSION The cellularity of the Bowman’s capsule,as well as histological alteration of kidney tubules,glomerular membranes,and liver tissues in diabetic rats after oral SBH resembled those of ND rats.Therefore,SBH exhibited a protective hepatorenal effect in a diabetic rat model.

Transcriptional reprogramming during human osteoclast differentiation identifies regulators of osteoclast activity

Enhanced osteoclastogenesis and osteoclast activity contribute to the development of osteoporosis,which is characterized by increased bone resorption and inadequate bone formation.As novel antiosteoporotic therapeutics are needed,understanding the genetic regulation of human osteoclastogenesis could help identify potential treatment targets.This study aimed to provide an overview of transcriptional reprogramming during human osteoclast differentiation.Osteoclasts were differentiated from CD14+monocytes from eight female donors.RNA sequencing during differentiation revealed 8980 differentially expressed genes grouped into eight temporal patterns conserved across donors.These patterns revealed distinct molecular functions associated with postmenopausal osteoporosis susceptibility genes based on RNA from iliac crest biopsies and bone mineral density SNPs.Network analyses revealed mutual dependencies between temporal expression patterns and provided insight into subtype-specific transcriptional networks.The donor-specific expression patterns revealed genes at the monocyte stage,such as filamin B(FLNB)and oxidized low-density lipoprotein receptor 1(OLR1,encoding LOX-1),that are predictive of the resorptive activity of mature osteoclasts.The expression of differentially expressed G-protein coupled receptors was strong during osteoclast differentiation,and these receptors are associated with bone mineral density SNPs,suggesting that they play a pivotal role in osteoclast differentiation and activity.The regulatory effects of three differentially expressed G-protein coupled receptors were exemplified by in vitro pharmacological modulation of complement 5 A receptor 1(C5AR1),somatostatin receptor 2(SSTR2),and free fatty acid receptor 4(FFAR4/GPR120).Activating C5AR1 enhanced osteoclast formation,while activating SSTR2 decreased the resorptive activity of mature osteoclasts,and activating FFAR4 decreased both the number and resorptive activity of mature osteoclasts.In conclusion,we report the occurrence of transcriptional reprogramming during human osteoclast differentiation and identified SSTR2 and FFAR4 as antiresorptive G-protein coupled receptors and FLNB and LOX-1 as potential molecular markers of osteoclast activity.These data can help future investigations identify molecular regulators of osteoclast differentiation and activity and provide the basis for novel antiosteoporotic targets.

Impairment of the Regulation of Gonadal Function in Channa punctatus by Metacid-50 and Carbaryl under Laboratory and Field Conditions

Regulation of gonadal function by gonadotropic hormone (GtH) and gonadotropin-releasing hormone (GnRH) in Channa punctatus was significantly affected by nonlethal levels of Metacid-50 and Carbaryl. Under laboratory conditions, the time-dependent decrease in serum GtH level was higher in Carbaryl-treated fish than in Metacid-50-treated fish. The situation was reversed in the field, with a higher inhibitory effect of Metacid-50 being recorded. On the other hand, pituitary GtH content and GnRH activity were inhibited to a greater extent by Metacid-50 than by Carbaryl under both field and laboratory conditions. The present findings highlight that even low doses of Metacid-50 and Carbaryl are effective enough to cause reproductive damage, as evidenced by homeostatic unbalance of the reproductive regulatory system. 1990 Academic Press. Inc.

GENISTEIN INHIBITS PROLIFERATION OF HUMAN ENDOMETRIAL ENDOTHELIAL CELL IN VITRO

Objective To explore the effect of genistein on proliferation of human endometrial endothelial cells(HEECs)and glandular epithelium.Methods In vitro HEECs and human endometrial cancer-1B cell(HEC-1B)were cultured with 0,1,10,50,100,and 200 μmol/L of genistein alone or indicated concentrations of genistein combined with 0.2 or 1 nmol/L 17β-estradiol(17β-E2).Cell proliferation was determined by[3H]-thymidine incorporation and cell cycle was measured by flow cytometry.Results After 96 hours of treatment,genistein inhibited the proliferation of HEECs in a dose-dependent manner.The stimulation index reduced from 100%(without genistein treatment)to about 1%(200 μmol/L genistein).HEECs were arrested at G1/0 and G2/M phase when treated with genistein for 96 hours.When the concentration of genistein was 200 μmol/L,the percentages of HEECs at G1/0,G2/M,and S phase were 96.0%,2.1%,and 1.9%,respectively.However,when HEECs were treated without genistein,the percentages of HEECs at G1/0,G2/M,and S phase were 76.7%,8.5%,and 14.7%,respectively.17β-E2 could not influence the effects of genistein on the proliferation of HEECs.Meanwhile,genistein could suppress the proliferation of HEC-1B.If the stimulation index of HEC-1B was defined as 100% when HEC-1B was treated with different doses of 17β-E2(without genistein),it was 67%,19%,as well as 32% when cell was supplemented with 200 μmol/L genistein combined with 0,0.2,or 1 nmol/L 17β-E2,respectively.Conclusion Genistein at the concentration of 200 μmol/L can sufficiently inhibit the proliferation of HEECs and endometrial glandular epithelium simultaneously in vitro.

Effect of Certain Toxicants on Gonadotropin-Induced Ovarian Non-Esterified Cholesterol Depletion and Steroidogenic Enzyme Stimulation of the Common Carp Cyprinus carpio in vitro

Isolated ovarian tissues from the common carp, Cyprinus carpio were incubated in vitro to obtain a discrete effect of four common toxicants of industrial origin, namely phenol, sulfide, mercuric chloride and cadmium chloride, on gonadotropin-induced alteration of nonesterified and esterified cholesterol and steroidogenic enzymes, △5-3β-HSD and 17β-HSD activity. Stage II ovarian tissue containing 30-40% mature oocytes were shown to be most responsive to gonadotropins in depleting only nonesterified cholesterol moiety and stimulating the activity of both. Safe doses of above mentioned toxicants when added separately to stage II ovarian tissue with oLH (1 μg/incubation) gonadotropin-induced depletion of nonesterified cholesterol and gonadotropin-induced stimulation of the activity of both enzymes was significantly inhibited. Esterified cholesterol remained almost unaltered. Findings clearly indicate the impairment of gonadotropin induced fish ovarian steroidogenesis by the four toxicants separately.

Immunocytochemical Localization of Glucagon and Insulin Cells in Eutropis carinata with Respect to Reproductive Cycle

The pancreas of Eutropis carinata was studied immunocytochemically for the presence and topographic distribution of glucagon and insulin cells during annual reproductive cycles. The study revealed paracrine arrangement of the two types of ceils. The immunolocalized pancreatic cells revealed significant differences in mean numbers during different periods of reproduction. The numbers of glucagon-immunoreactive （GIR） and insulin-immunoreactive （IIR） cells were highest during the recrudesceut period which corresponded with low serum glucose level. Between the two immunoreactive cells, the GIR cells were predominant. Morphological differences between the two cells were observed by electron microscopy after staining with uranyl acetate and lead citrate. Serum glucose showed that cyclic change was highest during the reproductive period.

17α-hydroxylase/17,20 carbon chain lyase deficiency caused by p.Tyr329fs homozygous mutation:Three case reports

p.Tyr329fs is a cytochrome P450c17 mutation among Chinese individuals.However,data on 17-α-hydroxylase deficiency caused by cytochrome P450c17 p.Tyr329fs homozygous mutation are lacking.This paper is a case report of three patients homozygous for p.Tyr329fs who were diagnosed with 17-α-hydroxylase deficiency between 2005 and 2019.CASE SUMMARY Case 1 presented with hypertension,hypokalemia,sexual infantilism and delayed bone age.The patient had a 46,XY karyotype,was homozygous for p.Tyr329fs and was recently treated with dexamethasone 0.375 mg qn.Case 2 presented with hypokalemia,sexual infantilism,osteoporosis and delayed bone age.The patient had a 46,XY karyotype,was homozygous for p.Tyr329fs and was treated with dexamethasone 0.75 mg qn at the last follow-up.Serum potassium and blood pressure could be maintained within normal range for cases 1 and 2.Case 3 presented with amenorrhea,sexual infantilism,osteopenia and delayed bone age.The patient had a 46,XX karyotype,was homozygous for p.Tyr329fs and was treated with dexamethasone 0.75 mg qn and progynova 1 mg qd.Outpatient follow-up revealed an adrenocorticotropic hormone(8 AM)of<5.00 pg/mL.CONCLUSION The homozygous p.Tyr329fs mutation usually manifests as a combined deficiency,and definitive diagnosis depends primarily on genetic testing.

The correlation between TNF-α,IL-6 in cervical mucous during follicular development and ovulation stimulation in different protocols

Objective:To investigate the correlation between TNF-α and IL-6 levels in cervical mucous during follicular development and ovulation stimulation in different protocols.Methods:36 infertile women were set up as experimental groups,divided into CC,HMG,IVF-ET group,each group consisted of 12 infertile women and 15 women with normal menstrual cycles were choiced as control group.Cervical mucous during follicular phase,luteal phase and ovulation phase were collected.TNF-a,IL-6 levels in cervical mucous were measured by radioimmunology assay(RIA).Follicular development were monitored by transvaginal ultrasonagraphy.Results:(1)TNF-α levels in cervical mucous of experimental groups and control group were periodically various among the reproductive cycle.It increased during follicular phase,reached to peak during ovulation phase,and decreased during luteal phase(P<0.05).IL-6 levels had no obvious periodical changes.(2)Compared with CC and control group,levels of TNF-α,IL-6 in HMG and IVF-ET group were significantly higher(P<0.05).(3)Levels of TNF-α and IL-6 in cervical mucous were positively correlated with the dominant follicle diameter(r=0.261,r=0.192 respectively,P<0.05).(4)TNF-α and IL-6 showed positive correlation in the reproductive cycle(r=0.782,P<0.05).Conclusions:(1)TNF-α level shows a cyclic change in the reproductive cycle and peaks during ovulation,whereas IL-6 level does not.(2)TNF-α and IL-6 may play a certain role in the process of follicular development and ovulation.(3)The levels of TNF-α and IL-6 are up-regulated by gonadotrophic hormone.(4)TNF-α and IL-6 may have coordination properties and participate in the same biological effects.

Effects of leptin and neuropeptide Y on function of human ovarian granulosa cells in vitro

Objective: To investigate the effects of leptin and neuropeptide Y on steroidogenesis of human ovarian granulosa cells in vitro. Methods: Human ovarian granulosa cells were isolated from follicular fluid obtained during oocyte retrieval of in vitro fertilization-embryo transfer program and cultured for 2 days with various concentration of leptin(1,10,100 ng/ml) or neuropeptide Y (1×10-6, 1×10-7, 1×10-8 mol/L) alone or both,or with the combination of human chorionic gonadotropin (hCG 0, 20 IU/L). The medium was collected for estradiol (E2) and progesterone (P) measurements. Results: (1)Whether hCG existed or not, the adding of leptin did not alter estradiol and progesterone production by human granulosa cells (P>0.05).(2) Only when the concentration of neuropeptide Y was at 1×10-7mol/L,estradiol level was lower than that in the control (P<0.05).(3) The levels of estradiol in neuropeptide Y (1×10-7mol/L) plus hCG group were significantly higher than those with neuropeptide Y alone(P<0.05). (4) In the absence of hCG, the levels of estradiol in neuropeptide Y (1×10-7mol/L)plus leptin (10 ng/ml) group were significantly higher than those with neuropeptide Y(1×10-7mol/L)alone(P<0.05).Conclusions: (1)Leptin alone produced no direct effect on secretion of E2 and P from granulosa cells in vitro.(2)Neuropeptide Y alone may inhibit the secretion of E2, but the inhibition would probably be blocked with the presentation of hCG.(3)Leptin probably blocked the inhibition of neuropeptide Y on E2 secretion, and this may indicate that there were some coordination between leptin and neuropeptid Y on the level of ovarian function.

Baseline moderate-range albuminuria is associated with protection against severe COVID-19 pneumonia

BACKGROUND Diabetes mellitus is considered a leading contributor to severe coronavirus disease 2019(COVID-19).AIM To characterize differences between hospitalized diabetic patients with vs without COVID-19,and parameters associated with COVID-19 severity for prediction.METHODS This case-control study included 209 patients with type 2 diabetic mellitus hospitalized at the Galilee Medical Center(Nahariya,Israel)and recruited between September 2020 and May 2021,65 patients with COVID-19 infection in dedicated wards and 144 COVID-19-negative patients in internal medicine wards hospitalized due to other reasons.Clinical parameters-including age,type of antiglycemic medications,presence of retinopathy,smoking history,body mass index(BMI),glycosylated hemoglobin,maximum neutrophil:lymphocyte ratio(NLR_(max)),C-reactive protein(CRP),estimated glomerular filtration rate(eGFR),and albumin(blood and urine)-were compared between the two primary patient groups,and then between COVID-19-negative patients hospitalized due to infectious vs non-infectious disease.Finally,we explored which parameters were associated with severe COVID-19 pneumonia.RESULTS COVID-19-negative patients were older(63.9±9.9 vs 59.8±9.2,P=0.005),and had longer duration of diabetes(P=0.031),lower eGFR(P=0.033),higher albumin(P=0.026),lower CRP(P<0.001),greater smoking prevalence(P<0.001),and more baseline albuminuria(54.9%vs 30.8%,P=0.005)at admission;70%of COVID-19 patients with albuminuria had moderate-range albuminuria(albumin:creatinine 30-300 mg/g).Most of the patients with albuminuria had chronic kidney disease stage II(CKD II).Oral antiglycemic therapies were not significantly different between the two groups.Multivariable logistic regression showed that higher BMI was significantly associated with severe COVID-19(OR 1.24,95%CI:1.01-1.53,P=0.04),as was higher NLR_(max)(OR 1.2,95%CI:1.06-1.37,P=0.005).Surprisingly,pre-hospitalization albuminuria,mostly moderate-range,was associated with reduced risk(OR 0.09,95%CI:0.01-0.62,P=0.015).Moderate-range albuminuria was not associated with bacterial infections.CONCLUSION Moderate-range albuminuria in COVID-19-positive diabetic patients with CKD II is associated with less severe COVID-19.Further studies should explore this potential biomarker for risk of COVID-19-related deterioration and early interventions.

Preliminary Results of DHEA in Poor Responders in IVF

Objective: Dehydroepiandrosterone (DHEA) has been previously reported to improve ovarian response in IVF for poor responders. Its effect remains uncertain. The purpose of this study was to evaluate the effect of DHEA given during two years in our unit, in order to build hypothesis for a future randomized controlled trial. Design: Cohort of exposed and non-exposed DHEA study. Patient(s): 224 patients with AMH under 1.6 ng/ml undergoing IVF with a short antagonist protocol adding FSH and LH rec were enrolled for 327 IVF cycles. 176 patients had been treated with DHEA before IVF, and 151 had not. Intervention(s): None. Main Outcome Measure(s): Cancellation cycle rate, amount of gonadotrophins used, estradiol level on day six of stimulation, number of oocytes retrieved, metaphase II oocytes, blastocysts and frozen embryos, clinical pregnancy rate and miscarriage rate. Results: The cancellation rate was significantly higher in the group without DHEA (29.8% versus 13.1%, p = 0.002). The amount of gonadotrophins used for stimulation, the estradiol level on day six of stimulation, the number of oocytes retrieved, metaphase II oocytes, blastocysts, frozen embryos and miscarriage rate were not different in both groups. The clinical pregnancy rate per included patient in the DHEA group was 17.1% versus 8.6% in the non-treated group (p = 0.02). Conclusion: DHEA appears to improve clinical pregnancy rate by decreasing the cancellation rate.

Pyogenic arthritis,pyoderma gangrenosum,and acne syndrome in a Chinese family:A case report and review of literature

BACKGROUND Pyogenic arthritis,pyoderma gangrenosum,and acne(PAPA)syndrome is a rare autosomal dominant genetic disease characterized by severe autoimmune inflammation,caused by mutations in the PSTPIP1 gene.Due to PAPA heterogeneous clinical manifestation,misdiagnosis or delayed diagnoses are difficult to avoid.With the use of whole-exome sequencing,we identified a missense mutation in the PSTPIP1 gene in a Chinese family.To the best of our knowledge,this is the first case of PAPA reported in China.CASE SUMMARY A 9-year-old boy suffered from recurrent aseptic pyogenic arthritis triggered by minor trauma or few obvious predisposing causes for more than 3 years.Pyogenic arthritis occurred every 3-5 mo,affecting his knees,elbows,and ankle joints.Treatments,such as glucocorticoids,antibiotics,even surgeries could alleviate joints pain and swelling to some extent but could not inhibit the recurrence of arthritis.Similar symptoms were present in his younger brother but not in his parents.According to the whole-exome sequencing,a missense mutation in exon 11 of the PSTPIP1 gene(c.748G>C;p.E250Q)was detected in the boy,his young-er brother and his father.Taking into account the similar phenotypic features with PAPA syndrome reported previously,we confirmed a diagnosis of PAPA syndrome for the family.CONCLUSION In this case,a missense mutation(c.748G>C;p.E250Q)in PSTPIP1 gene was identified in a Chinese family with PAPA syndrome.Previous studies emphasize the fact that PAPA syndrome is hard to diagnose just through the clinical manifestations owing to its heterogeneous expression.Genetic testing is an effectual auxiliary diagnostic method,especially in the early stages of pyogenic arthritis.Only if we have a deep understanding and rich experience of this rare disease can we make a prompt diagnosis,develop the best clinical treatment plan,and give good fertility guidance.

Association between fasting and postprandial triglyceride levels and carotid intima-media thickness in type 2 diabetes patients

OBJECTIVE: To investigate the association of fasting and postpiandial triglyceride with carotid intima-media thickness among type 2 diabetic patients in southeast China. METHODS: A total of 78 type 2 diabetic patients from southeast China were recruited. All the subjects underwent a standardized mixed-meal test (SMMT) and an oral glucose tolerance test. Carotid intima-media thickness (IMT) was measured by color doppler ultrasound examination. RESULTS: Postprandial triglyceride (PTG) after SMMT increased significantly in type 2 diabetic patients. The carotid IMT in patients with postprandial hypertriglyceridemia was significantly greater than that in patients with postprandial normal-triglyceridemia (0.90 mm vs 0.81 mm, P 0.05). CONCLUSION: The results suggest that postprandial hypertriglyceridemia may be an independent risk factor of early atherosclerosis in type 2 diabetes.

PARADOXICAL EFFECT OF A GnRH AGONIST ON STEROIDOGENESIS IN CULTURED MONKEY GRANULOSA CELLS

In this study we demonstrate: (i) The GnRH agonist exerts a direct dose-dependet stimulative effect on the aromatase activity and progesterone production in cultured monkey granulosa cells; (ii)the stimulative effect on steroidogenesis can be completely blocked by concomitant treatment with a GnRH antagonist, suggesting that the actions of GnRH are mediated through stringent stereospecific recongnition sites; (iii) in addition to the stimulative effect, the GnRH agonist in the presence of gonadotropins also exerts an inhibitory effect, even though the peptide by itself is more effective in the stimulation of steroidogenesis, and the stimulation of gonadotropin on steroidogenesis could be gradually restored by decreasing the concentration of the GnRH agonist in the culture; and (iv) paradoxical effect can also be observed in the presence of cAMP-inducing agents, suggesting that the inhibitory action of the peptide on gonadotropin-induced steroidogenesis is localized at a step distal to the stringent

STUDIES ON RELATIONSHIP BETWEEN Na, K-ATPase ACTIVITY AND SPERM CAPACITATION IN GUINEA PIG

In order to elucidate the biochemical mechanism of sperm capaeitation, the relationship between plasmalemma Na, K-ATPase, capacitation and acrosome reaction was investigated.Plasmalemma of guinea pig spermatozoa was isolated by sucrose gradient centrifugation for the determination of Na, K-ATPase activity. As far as the authors are aware, the Na, K-ATPase activity in plasmalemma of the guinea pig has never been detected. By treating sperm plasmalemma with 0.05% DOC (deoxycholate), enzyme activity could be quantitatively measured. After spermatozoa were incubated in capacitation medium for 8 h, Na, K-ATPase activity was found to be decreased to 35.6% as compared with that before incubation. The spermatozoa incubated for 10.5h in capacitationmedium containing 1 and 5μmol/L ouabain showed 46.5% and 64.4% acrosome reactions respectively, while the acrosome reaction of the control group was only 27.4%. The above results suggest that the decrease in the Na, K-ATPase activity of guinea pig spermatozoa may

VARIATIONS OF LECTIN-BINDING GLYCOPROTEINS IN EARLY PREGNANT RABBIT EMBRYO

In the present study, the quantitative and qualitative changes of three kinds of lectinbinding glycoproteins of early pregnant rabbit embryos (D4—D12) were analyzed. The technique of Western blot, as well as video densitometer scanning and its analytic software was used for analysis. Results found that there were five specific lectin-binding glycoproteins in Day 4 to Day 6 blastocyst fluids: one ConA-binding glycoprotein about 70 kD, two WGA-binding glycoproteins respectively about 42 kD and 25kD and two PNA-binding glycoprotein respectively about 180kD and 75kD. They disappeared immediately after implan tation, It is demonstrated that there are stage-specific glycoproteins in rabbit blastocyst fluid which might be relevant to the recognition of pregnancy and implantation.

DETECTION OF HUMAN TROPHOBLAST INTERFERON WITH ENZYME-LINKED IMMUNOSORBENT ASSAY

Although it is well known that luteal lifespan in early pregnancy is prolonged by human chorionic gonadotropin (hCG), recent evidence has suggested that proteins other than hCG, especially interferon-like substances resembling trophoblast protein-1 of domestic ruminants, may also be involved in the maintenance of corpus luteum.

Molecular determinants regulating the release of the egg during ovulation:Perspectives in piscine models

Reproduction in bony fish,specifically in the female teleost,encompasses well-defined stages,including growth and development,final oocyte maturation and finally ovulation-a pre-requisite for forming fertilizable female gametes.The morphological changes encountered during ovulatory response engage a multitude of endocrine,autocrine,and paracrine factors.These include gonadal steroids,growth factors,inflammatory cytokines,prostaglandins,leukotrienes and matrix metalloproteinases.In recent years,the active participation of prostaglandins and their receptors in LH-dependent ovulatory action has received much attention.Considerable evidence also supports the participation of nitric oxide/nitric oxide synthase(NO/NOS)unit in regulating meiotic maturation and ovulatory cascade.However,relatively less information is available on physiological relevance,the inter-relationship and the conjoined efforts of these molecular candidates to trigger ovulation in the fish ovary.The prime objective of the present article is to summarize the potential cross-talk between major molecular candidates downstream of LH action in the follicular layer.Besides,the intraovarian role of prostaglandin(PG)vis-`a-vis NO/NOS system on mammalian and teleost ovulatory response has been emphasised.

A Study of Protein and Glycoprotein Syntheses in Pre-implantation Mouse Embryos Using Mini-2D-Electrophoresis, Video Densitometer Scanning and Computer-Image Analysis

In the present study, proteins and glycoproteins of mouse embryos at 2-cell,morula and blastocyst stages were analyzed.The techniques of 35S-Met incorporation,ConA antiserum-precipitating ConA-binding proteins, mini-2D-electrophoresis, fluorography,video densitometer scanning and the computer-lmage system were used for analyses.Results of the investigations indicated that proteins and glycoproteins were synthesized by the embryos in a gradual increase manner from 2-cell to blastocyst. A relatively large amount of glycoproteins was synthesized during the morula and blastocyst stages.Two specific small glycoproteins respectively with molecular weights about 6500 and 9000 and PIs at 5.0 and 6.5 were apparently synthesized in the blastocyst but not in the 2-cell or the morula.

Comprehensive Metabolic Profiling of Modified Gegen Qinlian Decoction by Ultra-high-Performance Liquid Chromatography-Diode Array Detection-Q-Exactive-Orbitrap-Electrospray Ionization-Mass Spectrometry/Mass Spectrometry and Application of High-Performance

Objective:Gegen Qinlian decoction(GQD)is a classical traditional Chinese medicine formulation which has been used for almost 2000 years.At Guang'anmen Hospital,Beijing,a modified GQD version(m GQD)with seven instead of four herbal ingredients has been applied to treat Type 2 diabetes.Quality control is a crucial prerequisite for the therapeutic application of herbal medicines.For the identification of products derived from classical GQD,the Chinese Pharmacopeia requires the analysis of only three marker compounds.Because m GQD is a more complex mixture containing seven herbs and hundreds of constituents,the pharmacopoeia method for GQD is inadequate.Materials and Methods:A more comprehensive characterization of the formula's constituents has been developed using ultra-high-performance liquid chromatography-diode array detection(UHPLC-DAD)-Q-Exactive-mass spectrometry(MS)in electrospray ionization positive and negative mode.Moreover,a new method for the fingerprint analysis of m GQD via high-performance thin-layer chromatography(HPTLC)has been established.Results:Altogether,91 compounds have been assigned to their originating plants and 84 substances were identified either by comparison with authentic references or with data from the literature.The HPTLC method is based on the application of two different mobile phases and is able to detect both lipophilic and hydrophilic constituents of m GQD.Conclusions:The modified GQD was extensively characterized by UHPLC combined with DAD and Q-Exactive Orbitrap high-resolution MS detection,leading to the assignment and identification of compounds present in the decoction.In addition,a new method for the fingerprint analysis of the m GQD using HPTLC was established,which allows fast and simple identification of the herbal ingredients in the mixture.