OBJECTIVETo establish a simple but effective method of laser scanning confocal microscopic imaging for Ca2+ oscillations of pancreatic acinar cells in adult mice.

METHODSPancreatic acinar cells from adult Kunming mice were isolated acutely with collagenase, and then loaded with fluo-4-AM, a Ca2+ indicator. A laser scanning confocal microscope armed with 488 nm laser was employed to record the dynamic fluorescent signals in-time and synchronously while acetylcholine (ACh) was added in the pancreatic acinar cells.

RESULTS(1) The classic pancreatic acinar cell Ca2+ oscillations were induced by a certain concentration of ACh (100 nmol/L) successfully and steadily, which could be blocked by atropine completely. (2) Plasmic Ca2+ oscillations from different parts of one acinar cell were usually with different amplitudes and almost the same frequencies. But both of amplitudes and frequencies were different among different cells. (3) The acinar cell Ca2+ oscillations were induced by ACh in a concentration-dependent manner.

CONCLUSIONThe laser scanning confocal microscopic imaging for adult mouse pancreatic acinar cell Ca2+ oscillations was established successfully. The features of being easy to use, direct to see lively, high efficiency and good flexibility make it a popular tool for researchers to choose.

Acinar Cells ; chemistry ; Animals ; Calcium ; analysis ; Calcium Signaling ; Cells, Cultured ; Mice ; Microscopy, Confocal ; methods ; Pancreas ; cytology

Bacterial Typing Techniques ; methods ; Brucella ; classification

Objective To explore the correlation between hyperuricaemia and blood pressure, and blood lipid and glucose. Methods By using simple cluster sampling, 2 branch units from PetroChina Changqing Oilfield Company were selected, and all the 720 subjects with hyperuricaemia (HUA) were assigned to the HUA group; another 620 participants with normal uric acid (UA) level into the normal group. The correlation between blood uric acid and blood pressure,and blood lipid and glucose was assessed by Logistic regression. Results The odds ratio (OR) of those who had 1,2 or 3 abnormal status of hypertension,hyperlipidemia and impaired fasting glucose in the HUA group were much higher than the normal group (OR values were 4. 036,2. 562, and 4. 174, respectively). Logistic regression showed that male, systolic blood pressure ( SBP), GLU, total cholesterol ( TC), triglyceride ( TG), low-density lipoprotein cholesterol (LDL-C) were risk factors of H UA ( OR values were 7. 736,2. 309,1.721,2. 761 and 1. 411,respectively) ,while high-density lipoprotein cholesterol (HDL-C, OR = 0. 211 ) was a protective factor of HUA. Conclusions Gender,blood pressure and blood lipid may have correlation with blood UA. Multiple risk factors should be considered to improve the effectiveness of health education and health promotion.

BACKGROUND:Chinese medicine therapy is effective for patients with mild-to-moderate lumbar spinal stenosis and also has a better long-term effect than surgical patients, which can enhance perioperative efficacy and prevent spinal restenosis. OBJECTIVE:To evaluate the clinical efficacy of “Tongmai songjin yigu” massage therapy on lumbar spinal stenosis. METHODS:Eighty-four patients with degenerative lumbar spinal stenosis were randomly and equaly divided into treatment group and control group. Treatment group were treated with “Tongmai songjin yigu” massage therapy; patients in the control group were treated according to the guidelines for diagnosis and treatment of common disease in orthopedics and traumatology promulgated by China Association of Chinese Medicine, once a day, continuous for 2 weeks. RESULTS AND CONCLUSION:After treatment, the visual analog scale score in the treatment group was improved significantly (P < 0.05), and there was a significant difference in the Japanese Orthopaedic Association Scores on subjective symptom and daily life between the treatment and control groups (P < 0.05). The effective rate of the treatment group was higher than that of the control group (P< 0.05). These findings indicate that “Tongmai songjin yigu” massage therapy is better than traditional massage, but clinical large-sample studies are required.

Adult ; Carcinoma, Adenoid Cystic ; complications ; Female ; Humans ; Thyroid Gland ; pathology ; Thyroid Neoplasms ; secondary ; Ultrasonography

OBJECTIVETo study the influence of male age on the outcome of conventional IVF-ET.

METHODSBased on male age, 170 couples undergoing conventional IVF-ET were divided into three groups: <35 yr (n = 60), 35 -39 yr (n = 77) and > or = 40 yr (n = 33). We observed the rates of fertilization, cleavage, good quality embryo, implantation, clinical pregnancy and abortion in different groups.

RESULTSThere were no significant differences among the three groups in semen volume ([3.10 +/- 1.22] ml vs [2.84 +/- 1.05] ml vs [2.80 +/- 0.79] ml), sperm concentration ([54.23 +/- 26.07] x 10(6)/ml vs [60.27 +/- 24.80] x 10(6)/ml vs [60.21 +/- 27.42] x 10(6)/ml) and sperm viability ([53.93 +/- 13.25]% vs [56.10 +/- 16.58]% vs [51.82 +/- 15.45]%) (P>0.05). The men of the > or = 40 yr group showed a significantly lower percentage of grade a + b sperm ([40.97 +/- 11.91]%) than those of the <35 and 35 - 39 yr groups ([48.47 +/- 11.78]% and [46.84 +/- 13.51]%) (P<0.05), and morphologically normal sperm ([11.76 +/- 5.97]%) than those of the <35 yr group ([15.25 +/- 6.94]% (P<0.05). The rates of fertilization, cleavage, good quality embryo, implantation, clinical pregnancy were 81.52%, 82.61%, 52.33%, 18.06% and 33.33% in the > or = 40 yr group, with no significant differences from those of the <35 yr group (83.18%, 82.68%, 56.99%, 22.40% and 40.00%) and the 35 - 39 yr group (78.78%, 80.66%, 55.01%, 21.74% and 38.96%) (P>0.05), while the abortion rate was markedly increased in the > or = 40 yr group as compared with the <35 yr group (36.36% vs 8.33%, P>0.05).

CONCLUSIONIncreasing male age is related with decreasing percentages of progressively motile sperm and morphologically normal sperm, but not obviously with the rates of fertilization, good quality embryo, implantation, pregnancy and abortion.

Adult ; Age Factors ; Female ; Fertilization in Vitro ; Humans ; Male ; Middle Aged ; Paternal Age ; Pregnancy ; Pregnancy Outcome

OBJECTIVETo explore the correlation of males'age with sperm apoptosis, sperm DNA integrity and other seminal parameters.

METHODSWe collected 104 semen samples and divided them into three groups according to the males' age: <35 yr (n = 43), 35 -39 yr (n = 31), and > or = 40 yr (n = 30). Based on the WHO Laboratory Manual (4th ed), we detected the seminal parameters, calculated the percentage of apoptotic sperm by flow cytometry (FCM), determined sperm DNA integrity by Acridine orange staining, and compared the results among the three groups.

RESULTSThere were no statistically significant differences among the < 35 yr, 35 -39 yr and > or = 40 yr groups in semen volume ([2.87 +/- 0.89] ml vs [2.98 +/- 1.09] ml vs [2.65 +/- 0.95] ml), sperm concentration ([60.40 +/- 25.43] x 10(6)/ml vs [69.74 +/- 28.33] x 10(6)/ml vs [55.97 +/- 27.22] x 10(6)/ml) (P>0.05). The percentage of progressively motile sperm was significantly lower in the > or = 40 yr ([39.00 +/- 8.35 %) than in the <35 and 35 -39 yr groups ([48.73 +/- 9.89]% and [45.65 +/- 10.55]%) (P<.0.1), and so was the percentage of morphologically normal sperm in the > or = 40 yr than in the < 35 yr group ([11.11 +/- 8.26]% vs [16.43 +/- 8.75 ]%, P<0.01). The percentage of apoptotic sperm was markedly higher in the > or = 40 yr than in the <35 yr group ([11.82 +/- 5.77]% vs [7.04 +/- 3.50]%, P<0.01), while the sperm DNA integrity significantly reduced in the > or = 40 yr group ([75.52 +/- 10.60]%) as compared with the <35 yr ([86.55 +/- 5.60])% and 35 -39 yr group ( [81.39 +/- 8.94]%) (P<0.01). The males' age was correlated positively with the rate of sperm apoptosis (P<0.01), and negatively with sperm DNA integrity and the percentage of progressively motile sperm (P<0.01).

CONCLUSIONThe advance in males' age increases sperm apoptosis and reduces sperm progressive motility, normal morphology and DNA integrity.

Adult ; Age Factors ; Apoptosis ; genetics ; DNA ; Flow Cytometry ; Humans ; Infertility, Male ; genetics ; Male ; Sperm Count ; Sperm Motility ; Spermatozoa ; cytology

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology

OBJECTIVE: To explore the mechanism of bone marrow mesenchymal stem cells(BMSCs) in alleviating silica-induced lung injury in mice. METHODS: Ten specific pathogen free healthy male C57 BL/6 mice were selected for isolating BMSCs and bone marrow macrophages(BMDMs). Transwell chamber was used, BMDMs were inoculated onto the upper chamber and BMSCs in the lower chamber. We divided them into sequencing control group and silica(SiO_2) exposure group. All cells were pre-stimulated with 50 μg/L mass concentration lipopolysaccharide for 4 hours. In the SiO_(2 ) group, 250 mg/L mass concentration SiO_2 was added to the upper chamber of transwell and cultured for 16 hours. Total RNA was extracted from the BMSCs collected from the lower chamber. HiSeq/MiSeq high-throughput sequencing technology was used to detect the BMSCs RNA paired-end sequencing. Transcriptome sequencing data was obtained and bioinformatics analysis was performed. Another 12 specific pathogen free healthy male C57 BL/6 mice were randomly divided into control group and experimental group. All mice received one intra-tracheal injection of 20.0 μL(250 g/L mass concentration) of silica dust suspension. After 6 hours, the mice in the control group was given 500.0 μL of 0.9% sodium chloride solution and mice in the experimental group was given 500.0 μL of BMSCs suspension(cell density 1×10~9/L) by tail vein infusion.Mice were sacrificed 12 hours later. The relative mRNA expression of interleukin(IL)-1 Ra, IL-10, tumor necrosis factor stimulating gene 6(TSG-6) and prostaglandin E2(PGE2) in lung tissues of mice were measured by quantitative real-time PCR(q-PCR). Meanwhile, BMDMs and BMSCs transwell co-culture models were established. The cells were divided into 5 groups: BMSCs group, BMSCs+BMDMs group, BMSCs+BMDMs+ lipopolysaccharide(LPS) group, 50 mg/L SiO_2 group, and 100 mg/L SiO_2 group. After 16 hours of corresponding SiO_2 stimulation, BMSCs of each group were collected and the relative mRNA expression levels of IL-1 Ra, IL-10, TSG-6, and PGE2 in the cells were detected by q-PCR. RESULTS: Compared with sequencing control group, BMSCs co-cultured with SiO_2 had 19 genes up-regulated and 21 genes down-regulated, including 10 genes up-regulated for more than 2.0-fold. The relative mRNA expression of IL-1 Ra, IL-10, PGE2 and TSG-6 in the lung tissue of mice increased in the experimental group than that of the control group(all P<0.05). The relative mRNA expression of TSG-6 increased by 37.5 times higher than that of the control group. Compared with the BMSCs+BMDM+LPS group, the level of TSG-6 mRNA relative expression increased in both the 50 mg/L SiO_2 group and the 100 mg/L SiO_2 group(all P<0.05). CONCLUSION: TSG-6 could be the key factor of BMSCs that can attenuate silica-induced lung injury.

OBJECTIVETo observe the effects of forearms devascularization on the volt-ampere (V-A) characteristics of Neiguan (PC 6) and Taiyuan (LU 9) so as to investigate the relationship among functions of acupoints, qi and blood in human body and V-A characteristics of acupoints.

METHODSEither forearms of 23 healthy volunteers were devascularized and the changes of V-A characteristics in ipsilateral Neiguan (PC 6) and Taiyuan (LU 9) in normal and devascularization statuses were detected by home-made intelligent V-A detecting system.

RESULTSCompared with the normal status, both range-increasing and range-decreasing V-A areas of Neiguan (PC 6) and Taiyuan (LU 9) were significantly increased in devascularization status (P < 0.05, P < 0.01), and the change rates of these two V-A areas of Neiguan (PC 6) were larger than those of Taiyuan (LU 9) (both P < 0.05). The inertia areas of Neiguan (PC 6) but not Taiyuan (LU 9) became significantly larger after devascularization (P < 0.01).

CONCLUSIONThe changes in V-A characteristics of Neiguan (PC 6) and Taiyuan (LU 9) can reflect the alteration of local blood supply, and Neiguan (PC 6) is more sensitive to changes of blood flow than Taiyuan (LU 9).

Acupuncture Points ; Adult ; Blood Circulation ; Electrophysiology ; Female ; Forearm ; blood supply ; physiology ; Humans ; Male ; Young Adult