Follicle-stimulating hormone (FSH) is synthesized and secreted by the anterior pituitary, which binds to its receptors expressed on the membrane of Sertoli cells in the testis to bring about spermatogenesis. With the development of DNA sequencing technology, FSH SNPs rs10835638 and FSHR SNPs rs6165, rs6166, and rs1394205 were detected, which might directly affect the expression of FSH and activity of FSHR, resulting in male spermatogenic dysfunction. This review focuses on the relationship of FSH and FSHR gene polymorphisms with male infertility.
Follicle Stimulating Hormone ; genetics ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Receptors, FSH ; genetics ; Sertoli Cells ; Spermatogenesis ; Testis

OBJECTIVEMesenchymal stem cells (MSCs) from bone marrow are capable of differentiating into cells of different tissue lineages such as bone, cartilage, and adipose tissue and are the best candidates for tissue engineering. It is well accepted that umbilical cord blood (UCB) is a source for hematopoietic stem cells. However, controversy exists as to whether UCB contains MSCs and can serve as a source of MSCs. Therefore, the aim of this study was to explore the biological characteristics and inducing differentiation ability of in vitro expanded UCB MSCs.

METHODSUCB was collected on normal full term delivery of infants with informed consent (n = 35) obtained from the mothers. Mononuclear cells (MNCs) were isolated from UCB by gravity centrifugation and cultured with DMEM including 10% fetal bovine serum. The morphology was observed under microscope per day. Cytochemical staining was carried out and flow cytometry was used to examine the surface antigen phenotype. Fifth passage cells were transferred into a different medium and osteogenic differentiation, adipogenic differentiation, and neurogenic differentiation were assessed.

RESULTSMSCs could be isolated and cultured from MNCs of a few UCB sources. These cells displayed fibroblast-like morphology. They withstood over 20 passages without significant structural changes. These MSCs were negative for alkaline phosphatase (ALP) staining and positive for alpha-naphthol butyric acid esterase (NBE) staining. Expression of CD(29), CD(44)and CD(105), especially the human MSCs-specific markers SH-2 and SH-3 were observed, but CD(3), CD(14), CD(19), CD(34) and CD(45) could not be found, indicating that these cells were not of hematopoietic origin. Exposure of these MSCs to serum-free osteogenic condition, they could differentiate into bone cells and form mineralized matrix as evidenced by Alizarin red staining 2 weeks later. When these UCB-derived MSCs were cultured in adipogenic medium, morphologic changes in cells as well as the formation of neutral lipid vacuoles were noticeable as early as 1 week after induction and visualized by staining with oil-red O. Surprisingly, these MSCs were also able to differentiate into neuroglial-like cells. Morphology of these induced cells resembled that of neurons. Immunocytochemistry showed that they expressed Nestin and neuron-specific enolase (NSE), but not glial fibrillary acidic protein (GFAP).

CONCLUSIONUCB does contain MSCs. These MSCs, which are multipotent, could be isolated and cultured from a few UCB sources. UCB might serve as an alternative source of MSCs to bone marrow.

Alkaline Phosphatase ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Proliferation ; Centrifugation, Density Gradient ; Culture Media, Conditioned ; Fetal Blood ; cytology ; Flow Cytometry ; Histocytochemistry ; Humans ; Infant, Newborn ; Mesenchymal Stromal Cells ; metabolism ; Phosphopyruvate Hydratase ; metabolism

Acquired Immunodeficiency Syndrome ; epidemiology ; transmission ; Female ; Homosexuality, Female ; Humans ; Risk-Taking

To compare the growth characteristics of non-hematopoietic adult stem cells (NASC) derived from rat fetal blood and rat bone marrow in vitro, and to study the differentiation of these stem cells into neuron-like cells in vitro, the fetal blood of pregnant rats and bone marrow of adult rats were sterilely collected; mononuclear cells (MNC) were isolated by using standard Ficoll-hypague techniques and then cultured in DMEM/LG containing 10% fetal bovine serum (FBS). The acquired NASCs were subcultured for passage. The immunophenotype of NASCs was detected by flow cytometry. The expanded NASCs were induced to differentiate into neurons-like cells by beta-mercaptoethanol (beta-ME), dimethylsulfoxide (DMSO), butylated hydroxyanisole (BHA). The specific markers of these neuron-like cells were detected by immunocytochemistry. The results showed that two kinds of subcultured NASCs showed homogeneous spindle-shaped and expressed antigens CD44 and CD54, but did not expressed CD11b and CD45. The both induced cells were similar to neuron in morphology and were positive for nestin and neuron-specific enolase (NSE), but negative for glial fibrillary acidic protein (GFAP). It is concluded that no significant difference of NASCs derived from pregnant rat fetal blood and adult rat bone marrow found in cell morphology and biological characteristics. NASCs of both origins can be induced to differentiate into neuron-like cells, so fetal blood can be regarded as another resource of NASC.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; physiology ; Female ; Fetal Blood ; cytology ; Male ; Multipotent Stem Cells ; cytology ; physiology ; Neurons ; cytology ; Pregnancy ; Rats ; Rats, Wistar ; Stem Cells ; cytology

OBJECTIVETo find out the relationship between mutation of ATP7B gene promoter region and pathogenesis of Wilson disease(WD).

METHODSTwo of 48 WD patients presented C-->T base substitution mutations at the position -183. DNA sequences of the promoter region from normal and mutant samples were separated. The fragments containing the promoter region were cloned upstream of the luciferase. Luciferase activity was analyzed.

RESULTSThe luciferase activity of reporter gene containing normal sequence of ATP7B gene promoter region did not show significant difference as compared with that of reporter gene containing mutant promoter(n=3, P > 0.05).

CONCLUSIONNo influence of C-->T base substitution mutations on the activity of promoter was observed in study. The results suggest that WD pathogenesis relates little to the mutations of the promoter region in Chinese.

Adenosine Triphosphatases ; genetics ; Adolescent ; Adult ; Base Sequence ; Cation Transport Proteins ; genetics ; Cell Line, Tumor ; Child ; Copper-transporting ATPases ; DNA Mutational Analysis ; Female ; Hepatolenticular Degeneration ; genetics ; Humans ; Luciferases ; genetics ; metabolism ; Male ; Middle Aged ; Mutation ; Promoter Regions, Genetic ; genetics ; Young Adult

AIM:To investigate the effects of platelet-derived growth factor receptor α(PDGFRα) on melano-cyte apoptosis induced by hydrogen peroxide(H2O2). METHODS:Melanocyte PIGI was used as the research object. Af-ter exposed to H2O2at different concentrations,the cell viability was detected by MTT assay. The PIGI cells were transfec-ted with empty vector pCMV6 or PDGFRα over-expression vector pCMV6-PDGFRα. The transfection efficiency was deter-mined by RT-qPCR and Western blot. The effect of H2O2on the viability of the PIGI cells after over-expression of PDGFRα was measured by MTT assay. The cell apoptosis was analyzed by flow cytometry. The protein levels of p38, p-p38 and cleaved caspase-3 in the cells were detected by Western blot. DCDHF-DA was used to estemate the generation of reactive oxygen species (ROS) in the cells. RESULTS:The viability of PIGI cells decreased after exposed to H2O2(P<0.05), and the half maximal inhibitory concentration of H2O2was 0.7 mmol/L. Transfection with PDGFRα over-expression vector successfully induced high expression of PDGFRα at mRNA and protein levels in the PIGI cells,and increased the viability of the cells with H2O2treatment(P<0.05). Over-expression of PDGFRα decreased the apoptotic rate of PIGI cells trea-ted with H2O2(P<0.05),and the level of ROS in the cells(P<0.05). The protein levels of cleaved caspase-3 and p-p38 were also decreased (P <0.05). CONCLUSION:PDGFRα inhibits the apoptosis of melanocytes induced by H2O2,partially reverses the growth inhibition of melanocytes by H2O2,and decreases the ROS level. The mechanism may be related to regulating the protein levels of p-p38 and cleaved caspase-3 in the cells.

Pharmaceutical cocrystals is an advanced technology to improve the physicochemical and biological properties of drugs. However, there are few studies on the in vivo metabolism of pharmaceutical cocrystals. In this study, the pharmacokinetics of wogonin cocrystal in normal rats was further studied on the basis of the previous preparation of wogonin-aloperine cocrystal. Firstly, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for simultaneous determination of wogonin and its metabolite wogonoside in rat plasma, and to investigate the methodology. The method was applied to the pharmacokinetic study of wogonin-aloperine cocrystal (Wog-Alop) in rats. The results showed that wogonin and its metabolite wogonoside had a good linear relationship in the range of 1-800 ng·mL^-1, and the precision, accuracy, matrix effect and stability in this range met the requirements of biological analysis. Compared with direct administration of wogonin, the C_max of wogonin and its metabolite in rats increased to 7.44-fold and 9.15-fold, AUC_0-t increased to 1.67-fold and 3.72-fold, and oral bioavailability of wogonin increased to 187.66% after cocrystal administration. Wog-Alop cocrystal can significantly improve the C_max, AUC, and oral bioavailability of wogonin and its metabolite, which provides a new perspective for the clinical application of wogonin. This study was approved by the Experimental Animal Ethics Review Committee of Beijing University of Chinese Medicine (approval number: BUCM-2023032307-1148).

OBJECTIVETo investigate the protective effect of reactive oxygen species (ROS) scavenger, N-acetyl-L-cysteine (NAC), against H9c2 cardiomyocytes from injuries induced by chemical hypoxia.

METHODSH9c2 cells were treated with cobalt chloride (CoCl2), a chemical hypoxia-mimetic agent, to establish the chemical hypoxia-induced cardiomyocyte injury model. NAC was added into the cell medium 60 min prior to CoCl2 exposure. The cell viability was evaluated using cell counter kit (CCK-8), and the intercellular ROS level was measured by 2', 7'- dichlorfluorescein-diacetate (DCFH-DA) staining and photofluorography. Mitochondrial membrane potential (MMP) of the cells was observed by Rhodamine123 (Rh123) staining and photofluorography, and the ratio of GSSG/ (GSSG+GSH) was calculated according to detection results of the GSSG kit.

RESULTSExposure of H9c2 cardiomyocytes to 600 micromol/L CoCl2 for 36 h resulted in significantly reduced cell viability. Pretreatment with NAC at the concentrations ranging from 500 to 2000 micromol/L 60 min before CoCl2 exposure dose-dependently inhibited CoCl2-induced H9c2 cell injuries, and obviously increased the cell viability. NAC at 2000 micromol/L obviously inhibited the oxidative stress induced by CoCl2, decreased the ratio of GSSG/(GSSG+GSH), increased ROS level, and antagonized CoCl2-induced inhibition on MMP.

CONCLUSIONNAC offers obvious protective effect on H9c2 cardiomyocytes against injuries induced by chemical hypoxia by decreasing in the ratio of GSSG/(GSSG+GSH) and ROS level and ameliorating MMP.

Animals ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Embryo, Mammalian ; Free Radical Scavengers ; pharmacology ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Rats ; Reactive Oxygen Species ; metabolism

OBJECTIVETo observe the influences of seasonal factors on peripheral facial paralysis by acupuncture.

METHODSFour hundred cases of facial paralysis were divided into spring, summer, autumn and winter groups, 100 cases in each group. All these cases were treated by routine puncture. Fengchi (GB 20), Yifeng (TE 17), Qianzheng (Extra), Jiache (ST 6), and Dicang (ST 4), etc. were applied at affect side, once a day. 2 months observation was carried on to compare the clinical therapeutic effects and average courses. The facial symptoms, physical sign and functional activities were taken as observation indexes of therapeutic effect.

RESULTSThe effect rate was 78.0% (78/100) in spring group, 82.0% (82/100) in summer group, 89.0% (89/100) in autumn group and 92.0% (92/100) in winter group; the effect rate in summer or autumn group was superior to those in spring group and in summer group (all P < 0.05); the average course was (47.6 +/- 22.3) days in spring group, (43.7 +/- 18.4) days in summer group, (31.5 +/- 11.3) days in autumn group and (22.6 +/-9.2) days in winter group, indicating the significant differences between groups except that between spring and summer group (all P < 0.01). The cured and markedly effective rate was 80.1% (161/201) for wind cold type, 53.5% (61/114) for wind heat type, and 36.5% (31/85) for damp heat type, indicating that it of wind cold type was superior to that of wind heat type or damp heat type (P < 0.001, P < 0.05).

CONCLUSIONThe syndrome distribution and courses of peripheral facial paralysis are different in different seasons, hence, the diseases should be treated according to attack time and syndromes.

Acupuncture Therapy ; Adolescent ; Adult ; Facial Paralysis ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Seasons

OBJECTIVETo evaluate postoperative glottic area and vocal quality of three various surgical techniques for treating bilateral vocal cord paralysis, including laser arytenoidectomy (Group A, 24 cases), reinnervation of the posterior cricoarytenoid muscle by phrenic nerve (Group B, 9 cases) and arytenoidectomy accompanying lateral cordopexy by extralaryngeal approach (Woodman's procedure, Group C, 13 cases).

METHODS46 cases suffered from bilateral recurrent laryngeal nerve injury were included in our study. The pre-postoperative glottic measurement and vocal acoustic parameters were analyzed.

RESULTSThe decannulated cases in group A and group B and group C were 22, 8, 13 respectively. The post-operative mean maximal glottic area was (47.2 +/- 7.4) mm2, (78.3 +/- 16.0) mm2, (48.1 +/- 6.5) mm2 respectively. Group B cases glottic area was larger than that of group A and group C (t value were 4.46 and 3.85, P value were 0.000 and 0.001). No significant difference was found between group A and group C (t = 1.68, P = 0.101). After surgery, in group A, 17 cases voice quality was the same compared with that of before surgery, and 7 cases voice quality had become worse; In group B, the voice quality had become better in 5 cases, completely recovered in 1 case, and had not change in 3 cases; In group C, the voice quality had become deteriorated in 10 cases and no change in 3 cases. And in group B, ipsilateral diaphragm paralysis in 9 cases after surgery, whose vital capacity and forced vital capacity had decreased to 72%-84%, 76%-84% of that before the surgery respectively; and the diaphragm mobility had recovered by 35%-76% respectively, while vital capacity and forced vital capacity had become 93%-97%, 91%-98% of that before the surgery. In Group B, all cases' pulmonary function was normal half a year postoperatively.

CONCLUSIONSReinnervation of the posterior cricoarytenoid muscle by phrenic nerve seems to be best procedure with better post-operative voice and larger glottic area. Although the sufficient airway for decannulation can be acquired in Group A and Group C, but most of patients in Group A had pre-operative vocal level and badly abnormal in Group C.

Adult ; Aged ; Arytenoid Cartilage ; surgery ; Female ; Glottis ; physiopathology ; Humans ; Laser Therapy ; Male ; Middle Aged ; Phrenic Nerve ; surgery ; Treatment Outcome ; Vocal Cord Paralysis ; physiopathology ; surgery ; Voice Quality ; Young Adult