Administration, Inhalation ; Animals ; Erythrocyte Count ; Erythrocytes ; drug effects ; Female ; Formaldehyde ; toxicity ; Hemoglobins ; analysis ; Male ; Mice

In order to improve the expression level of segment of GABAA receptor a1 subunit in E. coli, growth conditions of the recombatant, which influence the final yield of protein expression, including growth medium, inoculation ratio, temperature, pH, rotation speed, inducing time and concentration of IPTG and so on, were studied in shaking flasks. The results indicated that, with 3% inoculation ratio, cultured 3.5 hours at 37℃, and then induced 5 hours by IPTG at 32℃, the yield of GABAA receptor protein was 95mg/L and the biomass was 3.25 g/ L. In contrast, using a 16 L stirred fermentor instead of shaking flasks, the highest level of the protein expression, 136mg/L with 4.95g/L of biomass, was achieved after fermenting 5.5 hours.

Adult ; Aged ; Aged, 80 and over ; Buruli Ulcer ; pathology ; surgery ; DNA, Bacterial ; analysis ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mycobacterium ulcerans ; genetics ; isolation & purification ; Tuberculosis, Cutaneous ; pathology

ObjectiveTo investigate the features of the cortisol ( COR),thyroid hormone and autonomic function of anxiety disorder,and to observe the relationships between clinical subtypes,insomnia,gender factors and the neuroendocrine and autonomic function.Methods60 anxiety disorder patients and 60 healthy controls were randomly selected from a three-grade class A hospital over the same period.Blood at 8:00am to test the plasma COR,T3,T4,TSH level and examined the heart rate variability (HRV) on the same day,the anxiety disorder patients also finish the self-rating anxiety scale (SAS).ResultsThe plasma COR,TSH level and LF/HF index were significantly higher( (531.58 ± 218.59 ) nmol/L,P＜ 0.0l ; (3.50 ± 1.35 ) MIU/L,P＜ 0.05 ;3.87 ± 3.37,P＜0.0l ),while the SDNN,LF,HF index were significantly lower in anxiety disorder than that in the control group (27.34±13.13,P＜0.01;338.72 ±396.34,P＜0.01;135.77 ±149.95,P＜0.01).The plasma COR,TSH level and LF/HF index were significantly higher(P ＜ 0.05,P＜0.01,P＜0.01 ),while the SDNN and HF index were significantly lower in panic disorder than that in the generalized anxiety disorder (P ＜ 0.01 ).The plasma COR,TSH level and LF/HF index were significantly higher,while the SDNN,LF,HF index were significantly lower in panic disorder than that in the control group (P＜0.01 ).The plasma COR was significantly higher,while the SDNN and HF index were significantly lower in generalized anxiety disorder than that of the control group (P＜0.01 ).The plasma COR level and LF/HF index were significantly higher (P＜0.01,P＜0.05),while the SDNN,LF,HF index were significantly lower in anxiety disorder with or without insomnia than that in the control group (P＜ 0.01 ).The plasma TSH level was significantly higher in anxiety disorder with insomnia than that in the control group (P＜ 0.05 ).There were no significant difference of the neuroendocrine and autonomic function between anxiety disorder with or without insomnia (P ＞ 0.05 ).The plasma COR,TSH level were significantly higher,while the SDNN,LF,HF index were significantly lower in the female group than that in the male group (P＜0.01).The total SAS scores were significantly positively correhted with plasma COR levels,and was significantly negatively correlated with SDNN,HF index in the anxiety group.ConclusionThere are neuroendocrine and autonomic nervous system dysfunctions in anxiety disorder,and these dysfunctions are related with clinical subtypes,insomnia,and gender.

Autosomal dominant polycystic kidney disease (ADPKD) is a most common inherited renal disease, about 50% with a family history, although the exact etiology not yet clear. To date, ADPKD, a multisystem disorder without effective preventive and therapeutic means, has been shown to be detrimental to human health. Recent studies show that severe oligoasthenozoospermia, necrospermia, immotile sperm, azoospermia, epididymal cyst, seminal vesicle cyst, and ejaculatory duct cyst found in male ADPKD patients may lead to male infertility, though the specific mechanisms remain unknown. Structural anomaly of spermatozoa, defect of polycystin, mutation of PKD genes, and micro-deletion of the AZF gene could be the reasons for the higher incidence of abnormal semen quality in male ADPKD patients. Assisted reproductive techniques can increase the chances of pregnancy, whereas the health of the offspring should be taken into consideration. This article presents an overview of reproductive issues concerning infertile male ADPKD patients from the perspective of the morbidity, pathophysiological mechanism, diagnosis, and management of the disease.
Cysts ; pathology ; Ejaculatory Ducts ; pathology ; Female ; Humans ; Infertility, Male ; physiopathology ; Kidney ; pathology ; Male ; Mutation ; Polycystic Kidney, Autosomal Dominant ; physiopathology ; Pregnancy ; Reproductive Techniques, Assisted ; Semen Analysis ; Spermatozoa ; pathology

Child ; Ectodermal Dysplasia ; diagnosis ; genetics ; Family Health ; Genes, Recessive ; genetics ; Humans ; Male ; Sex Factors

Objective To investigate the efficacy and side effects of 4?-adrenergic receptor(?- AR)antagonists in the treatment of chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS).Meth- ods Totally,325 patients(mean age,33.2 year;disease history,2.4 years)with CP/CPPS were randomly divided into Prazosin(n=95),Terazosin(n=78),Phenoxybenzamine(n=27),and Doxazosin mesylate controlled release tablets(n=72)groups.Some antibiotics and medications were used for allopathy.In addi- tion,53 cases with no?-AR antagonist treatment served as control group.The efficacy and side effects in all the patients were observed and recorded.The chronic prostatitis symptom index(CPSI)was used to evaluate the efficacy.Results In control group,22(41.5%)patients responded well to the treatment( CPSI mark drop≥5)and 31(58.5%)failed to respond to the treatment(CPSI mark drop＜5).In study group,199 (73.2%)patients responded well to the treatment,and 73(26.8%)failed.The difference in efficacy be- tween?-AR antagonists and placebo treatment was significant(P＜0.01).In study group,specifically,the effective rate was 55.6% in Phenoxybenzamine,78.2% in Terazosin,76.4% in Doxazosin mesylate con- trolled release tablets,and 71.6% in Prazosin groups.The main side effects of?-AR antagonists were postur- al hypotension(a rate of 23.2% for Prazosin,17.9% for Terazosin,22.2% for Phenoxybenzamine,and 8.3% for Doxazosin mesylate controlled release tablets)and dysfunction of ejaculation(only in Phenoxy- benzamine group with a rate of 51.9%).The rates of withdrawing treatment due to side effects were in turn 18.5% of Phenoxybenzamine,7.4% of Prazosin,5.1% of Terazosin,and 0% of Doxazosin mesylate con- trolled release tablets.Conclusions As essential medications for the treatment of CP/CPPS,?-AR antag- onists can relieve the clinical symptoms(dropping NIH-CPSI mark significantly),but some side effects should be considered when some medications are selected.

PURPOSE: Parathyroid injury is the major cause of post-thyroidectomy hypocalcemia. This prospective study aimed to define the relation between the preservation status of the parathyroid glands and post-thyroidectomy hypocalcemia as well as to determine whether the number of preserved parathyroid glands could be a guideline for the management of post-thyroidectomy hypocalcemia. METHODS: We analyzed a total of 76 cases of total thyroidectomy due to malignancy were analysed. Parathyroid gland status at the end of thyroidectomy was classified as preserved intact, discolored (color change or equivocal viability), or autotransplanted. Patients were grouped according to the number of intact preserved parathyroid glands into group 1 (more than two), group 2 (one), or group 3 (zero). The total and ionized serum calcium levels of each group were monitored for three days after surgery. RESULTS: The number of intact preserved parathyroid glands was not found to be closely related to serum calcium level, although ionized serum calcium level was significantly lower in group 1 than group 2 (P<0.001). Hypocalcemia was not found in group 1. Hypocalcemia was found in 31% (10/32) of group 2. Among the 32 patients of group 2, all 4 parathyroid gland were identified in 9 patients and 4 patients were shown to be hypocalcemic. Therefore, the incidence of hypocalcemia for patients in whom only one intact parathyroid was preserved and all 4 parathyroid were indentified was 44% (4/9). Hypocalcemia was transient in all patients. CONCLUSION: The parathyroid glands should be preserved as much as possible to minimize post-thyroidectomy hypocalcemia. The number of intact preserved parathyroid glands can be a good guideline for predicting post-thyroidectomy hypocalcemia and proper care management.
Calcium ; Humans ; Hypocalcemia* ; Incidence ; Parathyroid Glands ; Prospective Studies ; Thyroidectomy*

OBJECTIVETo construct the recombinant plasmid of RCAS1, to express and purify its fusion protein GST-RCAS1, and to investigate its biological function.

METHODSRCAS1 encoding gene was amplified by RT-PCR from total RNA extract of MCF-7 cells and was ligated with expression plasmid vector pGEX-2T by T4 DNA ligase after digested by the restricted endonucleases BamH I and EcoR I. Then the ligated products were inserted into competence JM109 E. Coli and the positive recombinants were identified by restriction endonuclease digestion assay and DNA sequencing. The GST-RCAS1 fusion protein expression was induced by IPTG in BL21 E. Coli and was purified with GST column and identified by SDS-PAGE and Western blotting with anti-GST monoclonal antibody, anti-RCAS1 (N-18) and anti-RCAS1 (C-20) polyclonal antibody. The apoptosis of activated T cells induced by GST-RCAS1 fusion protein was detected by flow cytometry with Annexin V and propidium iodide (PI) staining.

RESULTA 642 bp product was cloned by RT-PCR and the recombinant plasmid was constructed successfully. The GST-RCAS1 fusion protein was recognized by GST monoclonal antibody and RCAS1(N-18 and C-20) polyclonal antibody. FACS analysis showed that GST-RCAS1 fusion protein induced apoptosis in activated T cells.

CONCLUSIONThe recombinant plasmid of RCAS1 has been successfully constructed and the GST-RCAS1 fusion protein expressed and purified. The apoptosis inducing effect of GST-RCAS1 fusion protein on activated T cells is demonstrated.

Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Breast Neoplasms ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; Tumor Cells, Cultured

Chemical constituents of Swertia patens. The whole plant of air-dried Swertia patens was extracted with 90% EtOH. The water extract was suspended in H₂O and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isola- ted and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, ¹H-NMR, ¹³C- NMR). Eighteen compounds were isolated and elucidated as 3, 4-dihydro-1H,6H,8H-naptho [1,2-c:4,5-c', d'dipyrano-1, 8-dione (1), angelone (2), gentiogenal (3), erythricin (4), erythrocentaurin (5), gentianine (6), swertiakoside B (7), swertiamarin (8), 2'-O-actylswertiamarin (9), amarogentin (10), 1, 3, 5-trihydroxyxanthone (11), 1, 3-dihydroxy-5-methoxyxanthone (12), 1-hydroxy- 2, 3, 5-trimethoxyxanthone (13), gentiocrucine (14), 3-hydroxyphenylketone (15), n-hexacosyl ester 4-hydroxy-trans-cinnamate (16), n-hexacosyl ester 4-hydroxy-cis-cinnamate (17), and cholest-4-en-3-one (18). Compounds 1-7, 9-18 were obtained from S. patens for the first time.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Swertia ; chemistry