[Mechanism of Qingluo Tongbi Formula for regulating immune-bone erosion in rheumatoid arthritis].

OBJECTIVE: To explore the mechanism of Qingluo Tongbi formula for regulating "immune-bone erosion" in rheumatoid arthritis (RA). METHODS: Sixty-four RA patients were randomized into two groups to receive treatment with oral methotrexate or Qingluo Tongbi Formula for 12 weeks. Flow cytometry was used to analyze the changes in the percentages of CD3-CD19+, CD19+CD27 and CD19+BAFFR+B cell subpopulations in peripheral blood of the patients, and serum levels of B cell activating factor (BAFF), RANKL, RANK and osteoprotegerin (OPG) levels were detected using ELISA. Before and after the treatment, serum levels of ß-CTX, TRACP-5b, BGP, BALP, and PINP were measured with ELISA, and bone mineral density was determined with DXEA dual-energy X-ray absorptiometry. In the cell experiment, RAW264.7 cells were induced to differentiated into osteoclasts and treated with Qingluo Tongbi Formula at low-, moderate and high doses (125, 250 and 500 µg/mL, respectively) or with methotrexate (2 µg/mL) for 48 h, and the changes in the expression levels of RANKL, RANK, OPG and c-Fos were detected using Western blotting. RESULTS: The B cell subgroups in RA patients were correlated with the RANKL/RANK/OPG system. Treatment with Qingluo Tongbi Formula obviously down-regulated the percentages of the B cell subgroups, lowered serum levels of BAFF, ß-CTX and TRACP-5b, increased the levels of BGP, BALP and PINP, and improved lumbar bone density of RA patients (P<0.05); All these changes were significantly correlated with the regulation of B cell expressions (P<0.05). In RAW264.7 cells-derived osteoclasts, Qingluo Tongbi Formula significantly decreased the expressions of RANKL, RANK and c-Fos and increased the expression of OPG (P<0.05). CONCLUSION: Qingluo Tongbi Formula inhibits bone erosion in RA possibly by regulating B cell subset percentages and BAFF expression and inhibiting osteoclast differentiation via the RANKL/RANK/OPG pathway.

[Bushen Huoxue Fang improves recurrent miscarriage in mice by down-regulating the JAK2/STAT3 pathway].

OBJECTIVE: To investigate the efficacy of Bushen Huoxue Fang (BSHXF, a traditional Chinese medicine formula) for improving recurrent spontaneous abortion (RSA) in mice and the role of tyrosine kinase (JAK2) and transcriptional activator (STAT3) signaling pathway in its therapeutic mechanism. METHODS: Female CBA/J mice were caged with male DBA/2 mice to establish RSA mouse models, which were randomly divided into model group, dydrogesterone group and BSHXF group, with the female mice caged with male BALB/c mice as the control group (n=6). From the first day of pregnancy, the mice were subjected to daily intragastric administration of BSHXF, dydrogesterone, or distilled water (in control and model groups) for 12 days. After the treatments, serum levels of antithrombin III (AT-III), activated protein C (APC), tissue plasminogen activator (t-PA), progesterone, human chorionic gonadotropin (HCG), and estradiol (E2) were detected in each group using ELISA. HE staining was used to observe the morphological changes of the endometrium of the mice. Western blotting was performed to determine the expressions of p-JAK2, p-Stat3 and Bcl-2 in the placenta of the mice. RESULTS: Compared with the control mice, the mouse models of RSA showed a significantly increased embryo loss rate with decreased serum levels of AT-III, T-PA, progesterone, APC and HCG, increased placental expressions of p-JAK2, p-STAT3 and Bax, and decreased expression of Bcl-2 (P < 0.05). Treatments with BSHXF and dydrogesterone both increased serum levels of AT-III, t-PA and HCG in the mouse models; Serum APC level was significantly reduced in BSHXF group and serum progesterone level was significantly increased in dydrogesterone group (P < 0.05). CONCLUSION: BSHXF can improve the prethrombotic state and inhibit cell apoptosis by downregulating the JAK2/STAT3 pathway to increase the pregnancy rate in mouse models of RSA.

[Association study between the KCNE family gene polymorphisms of potassium channel gene and the susceptibility of atrial fibrillation].

Objective: To investigate the relationship between KCNE family gene polymorphisms of potassium channel gene and the susceptibility of atrial fibrillation (AF). Methods: In the case-control study, a total of 648 subjects were studied, of which 338 patients with atrial fibrillation were selected from the Department of Cardiovascular Medicine, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2019 to December 2019, and 310 healthy people were selected from the physical examination population during the same period. DNA sequencing technology and polymerase chain reaction (PCR) were used to detect the genotype and allele frequency of rs1805127 of KCNE1, rs9984281 of KCNE2, rs9516, rs626930 of KCNE3 and rs12621643 of KCNE4. Results: The ages of subjects in atrial fibrillation group and control group were (69±13) and (73±8) years, respectively (P=0.077). Men subjects accounted for 57.70% (195 men) and 40.00% (124 men) in the two groups, respectively (P=0.092). The distribution frequencies of the allele C at rs1805127 of gene KCNE1, the allele A at rs9984281 of gene KCNE2 and the allele G at rs12621643 of gene KCNE4 were significantly different between groups (P<0.05). After adjustment for sex, smoking, hypertension, cardiac insufficiency and other factors, it was found that the increase in the frequency of the above three loci would increase the risk of atrial fibrillation (rs1805127 OR=7.064, 95%CI:1.559-31.997; rs9984281 OR=4.210, 95%CI:1.118-15.850; rs12621643 OR=2.679, 95%CI:1.025-6.998). Conclusion: The rs1805127 of KCNE1, the rs9984281 of KCNE2,the rs12621643 of KCNE4 were significantly associated with the susceptibility to atrial fibrillation.

[Transmission chains of local epidemic of COVID-19 caused by 2019-nCoV Delta variant in Zhenhai district, Ningbo].

Objective: To investigate the local epidemic of COVID-19 caused by 2019-nCoV Delta variant in Zhenhai district of Ningbo, identify the transmission chain and provide reference for the prevention and control of COVID-19 epidemic. Methods: The incidence data of COVID-19 in Zhenhai from 6 to 18 December, 2021 were collected in field investigation. Field epidemiological investigation was conducted to understand the epidemiological characteristics of COVID-19 cases and analyze the transmission chains. Results: The first case might be infected with 2019-nCoV through direct or indirect exposure when passing through a medium-risk area, then a family cluster was caused, and the epidemic spread through close contacts of family members with others such as work, daily life, and moxibustion. The epidemic lasted for 14 days, and 74 confirmed COVID-19 cases were reported. The median incubation period was 4.0(3.0,5.8)d. All the cases were in a chain of transmission for more than 6 generations, and the intergenerational interval was 3.5(2.0,5.3)d. The gene sequencing result indicated that the pathogen was Delta AY.4 variant of 2019-nCoV. Both the epidemiological investigation and the gene sequencing results supported that the local COVID-19 epidemic in Zhenhai was associated with the COVID-19 epidemic in Shanghai. Conclusions: The transmission chain of this epidemic was clear. Delta AY.4 variant has obvious characteristic to cause case clusters in families, places with poor ventilation, and residential communities. It is suggested to strengthen the health management in key areas and key populations, and increase the frequency of nucleic acid testing.

[Correlation analysis of cervical spine dysfunction, pain and muscle strength in office workers].

Objective: To expore the correlation between neck disability, neck pain and muscle strength in cervical pondylosis of office worker, and to provide scientific basis for the prevention and treatment of cervical spondylosis. Methods: In April 2021 ,234 patients with cervical spondylotic myelopathy treated in the Subsidiary Rehabilitation Hospital of Fujian University of Traditional Chinese Medicine from April 2015 to April 2017 were selected, the correlation between Neck Disability Index (NDI) score, neck pain and muscle strength was analyzed using the Spearman rank correlation method. Mann-Whitney U test was used to compare the difference of maximum muscle strength of isometric contraction. Results: NDI score was negatively correlated with neck flexion, extension, and muscle strength in the left and right flexion directions (r(s)=-0.164, -0.169, -0.222, -0.176, P=0.012, 0.010, 0.001 , 0.007). In mild and moderate functional disorder patients, the muscle strength in flexion, extension and left and right flexion direction was greater, the difference was statistically significant (P <0.01). Conclusion: There is a negative correlation between cervical functional disorder and cervical muscle strength in office workers, suggesting that strengthening cervical muscle strength may be a way to improve cervical spine function.

Effect of chromium methionine supplementation on lactation performance, hepatic respiratory rate and anti-oxidative capacity in early-lactating dairy cows.

Chromium may regulate dairy cow metabolism; a chelated formation of chromium methionine (Cr-Met) is available to the feed industry. The objective of this study was to investigate the effect of Cr-Met supplementation on lactation performance, hepatic respiratory rate and anti-oxidative capacity in early-lactating Holstein dairy cows. 64 multiparous cows were assigned to 16 blocks based on parity and milk yield and then the four cows in a block were randomly allocated to four treatment groups with 0, 4, 8 or 16 g/d of Cr-Met per cow supplemented to a basal diet. Cows were moved from an open dry lot to a naturally ventilated tie stall barn 2 weeks before treatment to adapt to this facility, fed and milked at 0630, 1400, and 1930 h every day. The experiment lasted for 12 weeks. Milk yield and composition were recorded weekly. Dry matter intake was measured every 2 weeks for a total of six times throughout the trial. The plasma variables were measured in weeks 4, 8 and 12 of the experiment. Supplementation of Cr-Met did not affect DM intake of cows. As the supplementation of Cr-Met increased, yields of milk, fat, energy corrected milk (P < 0.01) and lactose (P = 0.01) increased in a linear manner. In terms of plasma variables, insulin concentration decreased in a linear manner with Cr-Met supplementation. As for variables relating to hepatic respiration rate, concentrations of pyruvate and NAD in the plasma were increased in quadratic manners, and lactic dehydrogenase activity was linearly increased as Cr-Met feeding levels increased. Moreover, plasma glutathione peroxidase and superoxide dismutase activity were increased in a linear manner. In conclusion, our study suggested that Cr-Met supplementation improved lactation performance of early-lactating dairy cows through enhancing antioxidant capacity and hepatic cellular respiration.

[The role of hypothalamus polycomb gene methylation in bisphenol A exposure during pregnancy and premature puberty in female offspring].

Objective: To explore the role of hypothalamus Polycomb Group (PcG) gene (Eed, Ezh) methylation in the relationship between bisphenol A (BPA) exposure during pregnancy and premature puberty in female offspring. Methods: A total of 40 pregnant CD-1 mice were randomly and averagely assigned into four groups: control group (corn oil) and low, middle and high BPA-exposed groups (the poisonous doses were 8 mg/kg, 40 mg/kg, and 200 mg/kg, respectively) by random number table method. Each group was administered by gavage from gestational day (GD) 1 to 18. The vaginal opening of female offspring was observed from postnatal day (PND) 21 to 33. All female offsprings were sacrificed, and hypothalamus was remained on the PND 34. The methylation levels of Eed and Ezh in the hypothalamus were measured. The early puberty of CD-1 mice was evaluated by the rate of vaginal opening in advance, initial time of vaginal opening, the first estrus occurrence and vaginal opening days in advance. The path model was used to explore the role of Eed and Ezh gene methylation in the early puberty of female offspring with maternal BPA exposed including the number of days of vaginal opening in advance as a dependent variable and BPA exposure as an independent variable. Results: The rate of vaginal opening on the 28 day in each maternal BPA-exposure group [low, middle and high BPA-exposed groups were 40.00% (29/72), 47.62% (25/53) and 37.84% (20/53), respectively] was higher than that rate in the control group [14.06%(9/64)]. Similarly, the P(50)(P(25), P(75)) values of initial time of vaginal opening in low, middle and high BPA-exposed group were 28 (26, 30), 28 (26, 29), 28 (26, 30) days, respectively and the P(50)(P(25), P(75)) values of the first estrus occurrence in low, middle and high BPA-exposed group were 31 (27, 32), 30 (27, 31), 31 (28, 33) days, respectively, which were earlier than those in the control group [initial time of vaginal opening was 30(28, 31) days, and the first estrus occurrence was 32(30, 33) days] (all P values<0.05). Compared with the control group (the methylation levels of Eed1, Eed2, Ezh2 were 1.47%, 1.26%, 2.56%, respectively), the methylation levels of Eed1 (1.61%-1.82%), Eed2 (1.36%-1.43%) and Ezh2 (2.87%-3.05%) in female offspring were significantly higher in BPA-exposed groups (all P values<0.05). The results of path model analysis showed that BPA had no direct influence on puberty in advance, but had an indirect effect on puberty in advance (indirect effect path coefficient was 0.045 and 0.142, respectively) by mediating methylation of Eed2, and Ezh2. Conclusion: Early puberty in female offspring induced by maternal exposure to BPA during pregnancy through the increased methylation levels of hypothalamus PcG gene (Eed, Ezh) in female offspring.

Maresin 1 attenuates neuroinflammation in a mouse model of perioperative neurocognitive disorders.

BACKGROUND: Resolution of inflammation is an active and dynamic process after surgery. Maresin 1 (MaR1) is one of a growing number of specialised pro-resolving lipids biosynthesised by macrophages that regulates acute inflammation. We investigated the effects of MaR1 on postoperative neuroinflammation, macrophage activity, and cognitive function in mice. METHODS: Adult male C57BL/6 (n=111) and Ccr2RFP/+Cx3cr1GFP/+ (n=54) mice were treated with MaR1 before undergoing anaesthesia and orthopaedic surgery. Systemic inflammatory changes, bone healing, neuroinflammation, and cognition were assessed at different time points. MaR1 protective effects were also evaluated using bone marrow derived macrophage cultures. RESULTS: MaR1 exerted potent systemic anti-inflammatory effects without impairing fracture healing. Prophylaxis with MaR1 prevented surgery-induced glial activation and opening of the blood-brain barrier. In Ccr2RFP/+Cx3cr1GFP/+ mice, fewer infiltrating macrophages were detected in the hippocampus after surgery with MaR1 prophylaxis, which resulted in improved memory function. MaR1 treatment also reduced expression of pro-inflammatory cell surface markers and cytokines by in vitro cultured macrophages. MaR1 was detectable in the cerebrospinal fluid of older adults before and after surgery. CONCLUSIONS: MaR1 exerts distinct anti-inflammatory and pro-resolving effects through regulation of macrophage infiltration, NF-κB signalling, and cytokine release after surgery. Future studies on the use of pro-resolving lipid mediators may inform novel approaches to treat neuroinflammation and postoperative neurocognitive disorders.

miR-132 weakens proliferation and invasion of glioma cells via the inhibition of Gli1.

OBJECTIVE: Transcriptional factor Gli1 in Hedgehog signal pathway facilitates epithelial mesenchymal transition (EMT) and is associated with invasion or proliferation of multiple tumor cells. The previous study showed the correlation between miR-132 down-regulation and glioma pathogenesis. We investigated the role of miR-132 in mediating Gli1 expression and in affecting proliferation or invasion of glioma cells. PATIENTS AND METHODS: Dual luciferase reporter gene assay was used to confirm the targeted regulation between miR-132 and Gli1. Tumor tissues at different pathological grades (grade II, III and IV) were collected from glioma patients, in parallel with brain tissues from contusion surgery. The expression of miR-132 and Gli1 was measured by RT-PCR. Glioma cell line U251 was treated with miR-132 or si-Gli1 followed by measuring the expression of Gli1, E-cadherin, Vimentin and Cyclin D1. In addition, flow cytometry and transwell assay were performed to evaluate cell invasion potency. RESULTS: Bioinformatics analysis showed the complementary binding sites between miR-132 and 3'-UTR of Gli1 mRNA. Transfection of miR-132 mimic significantly reduced luciferase activity, indicating the targeted regulatory relationship between miR-132 and Gli1 mRNA. Compared with control group, miR-132 expression was decreased and Gli1 level was elevated in glioma tissues, both of which were correlated with the pathological grade. Transfection of miR-132 mimic or si-Gli1 remarkably suppressed the expression of Gli1, Vimentin or Cyclin D1 in U251 cells, up-regulated E-cadherin expression, suppressed cell proliferation and invasion. CONCLUSIONS: Our data indicated that over-expression of miR-132 could inhibit proliferation or invasion of glioma cells via targeted inhibition of Gli1 expression.

[A brief textual research on Mei shi fang (Prescriptions of Master Mei) and Yi men fang (Prescriptions of Medical Professionals)].

Mei shi fang (Prescriptions of Master Mei) cited in Zheng lei ben cao (Classified Syndromes Materia Medica) and Yi men fang (Prescriptions of Medical Professionals) cited in Yi xin fang (Ishinpo) are actually the same book with different titles, which is actually called Yi men mi lu (Secret Records of Medical Professionals) with 5 volumes or the 5-volume Mei chong xian fang (Prescriptions of Mei Chongxian) approximately compiled during the years of Zhenyuan to Yuanhe reigns of the Tang Dynasty (785-820) written by Mei Chongxian, a Tang taoist. Mei Chongxian and Mei Biao, the author of Shi yao er ya (Dictionary of Mineral Medicines), might probably be the same person. According to the citations of Yi xin fang and Zheng lei ben cao, Yi men mi lu should be a comprehensive medical book including theories, methods, prescriptions and medications, recorded classical prescriptions of predecessors extensively, with some innovations and high clinical and documentary significance.

The anti-diabetic drug exenatide, a glucagon-like peptide-1 receptor agonist, counteracts hepatocarcinogenesis through cAMP-PKA-EGFR-STAT3 axis.

Epidemiological studies have demonstrated a close association of type 2 diabetes and hepatocellular carcinoma (HCC). Exenatide (Ex-4), a potent diabetes drug targeting glucagon-like peptide-1 receptor (GLP-1R), is protective against non-alcoholic fatty liver disease (NAFLD). However, the Ex-4 function and GLP-1R status have yet been explored in HCC. Herein we investigated the effect of Ex-4 in diethylnitrosamine (DEN)-treated mice consuming control or high-fat high-carbohydrate diet. Administration of Ex-4 significantly improved obesity-induced hyperglycemia and hyperlipidemia and reduced HCC multiplicity in obese DEN-treated mice, in which suppressed proliferation and induced apoptosis were confined to tumor cells. The tumor suppression effects of Ex-4 were associated with high expression of GLP-1R and activation of cyclic AMP (cAMP) and protein kinase A (PKA). Importantly, Ex-4 also downregulated epidermal growth factor receptor (EGFR) and signal transducer and activator of transcription 3 (STAT3), which lie downstream of cAMP-PKA signaling, resulting in suppression of multiple STAT3-targeted genes including c-Myc, cyclin D1, survivin, Bcl-2 and Bcl-xl. The growth inhibitory effects of Ex-4 were consistent in GLP-1R-abundant hepatoma cell lines and xenograft mouse model, wherein both PKA and EGFR had obligatory roles in mediating Ex-4 functions. In addition, Ex-4 also effectively suppressed inflammatory and fibrotic phenotypes in mice fed with methionine-choline-deficient (MCD) diet and choline-deficient ethionine-supplemented (CDE) diet, respectively. In summary, Ex-4 elicits protective functions against NAFLD and obesity-associated HCC through cAMP-PKA-EGFR-STAT3 signaling, suggesting its administration as a novel approach to reduce HCC risk in diabetic patients.

Maternal obesity during pregnancy is negatively associated with maternal and neonatal iron status.

BACKGROUND/OBJECTIVES: Obesity among pregnant women may adversely affect both maternal iron status throughout pregnancy and placental transfer of iron. The objective of this study was to determine the association of maternal body mass index (BMI) with (1) maternal iron status and inflammation in mid and late pregnancy, (2) the change in maternal iron status throughout pregnancy and (3) neonatal iron status. SUBJECTS/METHODS: We examined longitudinal data from 1613 participants in a pregnancy iron supplementation trial in rural China. Women with uncomplicated singleton pregnancies were enrolled in the early second trimester of pregnancy and followed through parturition. Maternal blood samples obtained at enrollment and in the third trimester and cord blood samples were analyzed for a range of hematological and iron biomarkers. RESULTS: There was a negative association between maternal BMI and iron status at enrollment (transferrin receptor (sTfR): r=0.20, P<0.001; body iron (BI): r=-0.05; P=0.03). This association was markedly stronger among obese women. Maternal BMI was positively associated with maternal inflammation (C-reactive protein: r=0.33, P<0.001). In multiple linear regression models, maternal BMI was negatively associated with neonatal iron status (cord serum ferritin: -0.01, P=0.008; BI: -0.06, P=0.006) and associated with a lower decrease in iron status throughout pregnancy (sTfR: -4.6, P<0.001; BI: 1.1, P=0.004). CONCLUSIONS: Maternal obesity during pregnancy may adversely affect both maternal and neonatal iron status, potentially through inflammatory pathways.

[Outcomes of total deafness type of idiopathic suddendeafness in different ages].

Objective:This retrospective study was aimed to investigate the characteristics of hearing recovery in the complete deafness type of SSNHL(≥81 dBHL) in patients with different ages.Method:Clinical outcomes of 179 total deafness type of idiopathic sudden deafness were compared.Patients were divided into 5 groups according to age,they were,pediatric group(13 years or less),youthful group(14-44 years),middle-aged group(45-59 years),presenium group(60-74 years),senectitude group(75 years or higher).Patients were divided into 3 groups according to the initial degree of hearing loss: 81 dB group (81-89 dBHL),90 dB group(90- 99 dBHL),100 dB group(100 dBHL or higher).Routine comprehensive treatment including corticosteroids,the inner ear microcirculation improvement drugs,neurotrophic drugs,saturationoxygen and hyperbaric oxygen therapy,etc.was applied.Patients were treated in accordance with the age and body weight.Result:The percentage of youthful group(83/179,46.4%) was highest(P<0.05),middle-aged group(57/179,31.8%)followed(P<0.05),presenium group(26/179,14.5%)was lower(P<0.05),pediatric group(8/179,4.5%) and senectitude group(5/179,2.8%)were the lowest.No a complete recovery in either pediatric group or senectitude group.A complete recovery was rare in the other groups.Recovery rate of the different aged groups was similar(P>0.05).The percentage of 100 dB group(108/179,60.3%) was highest(P<0.05).The percentage of 81 dB group(39/179,21.8%)was similar to 90 dB group(32/179,17.9%)(P>0.05).Recovery rate was similar in 81 dB group(25/39,64.1%)and 90 dB group(18/32,56.2%)(P>0.05).Recovery rate of both 81 dB group and 90 dB group were greater than 100 dB group(24/108,22.2%)(P<0.05).The 100 dB group reduced the satisfactory recovery effects.There were no differences in the proportion of the patients with dizziness(95/179,53.1%)and without dizziness(84/179,46.9%)(P>0.05).Recovery rate of patients without dizziness(43/84,51.2%) was greater than with dizziness(24/95,25.3%)(P<0.05).The percentage of the patients without dizziness(31/39,79.5%)in 81 dB group was the highest(P<0.05),90 dB group(18/32,56.2%)followed(P<0.05).The percentage of the patients with dizziness in 100 dB group(73/108,67.6%)was highest(P<0.05).Recovery rate was similar in the patients without dizziness of 81 dB group(21/31,67.7%)and 90 dB group(11/18,61.1%)(P>0.05).Recovery rate of the above two groups was greater than that of 100 dB group(11/35,31.4%)(P<0.05).Conclusion:Recovery rate of the different aged groups was similar.The percentage of the patients with dizziness in 100 dB group was highest.Initial hearing threshold in excess of 100 dB reduced the satisfactory recovery in patients with total deafness type of SSNHL.Our results provided a good reference for other clinicians.

An improved dosage regimen of sertraline hydrochloride in the treatment for premature ejaculation: an 8-week, single-blind, randomized controlled study followed by a 4-week, open-label extension study.

WHAT IS KNOWN AND OBJECTIVE: This study aimed at evaluating the safety and efficacy of an improved dosage regimen of sertraline in patients with premature ejaculation (PE) and to examine whether the premature ejaculation diagnostic tool (PEDT) can be used as a measure of treatment response in these patients. METHODS: A total of 218 PE patients were randomized into control (n = 61) and treatment (n = 157) groups to receive mycelium of cordyceps sinensis C4 and sertraline 50 mg daily for 8 weeks, respectively. Following this blinded stage, sixty-three patients chose to take sertraline 100 mg daily for an additional 4-week period, and 80 other patients continued treatment with sertraline 50 mg. Main outcome measures include intravaginal ejaculatory latency time (IELT), PEDT score and Clinical Global Impression of Change (CGIC) score. RESULTS: At weeks 4 and 8, mean IELT of patients who subsequently chose to take 100 mg of sertraline was significantly lower than that of patients who continued taking 50 mg of sertraline, although the IELT value was comparable between the two groups of patients at baseline. However, with an additional 4-week treatment, the mean IELT increased significantly more in the 100-mg group than in the 50-mg continuation group. Similar results were also obtained in the analyses of the PEDT and CGIC scores. Both regimens were well tolerated, and relapse rate did not differ significantly between the two groups. WHAT IS NEW AND CONCLUSION: These findings suggest that PE patients not responding to an 8-week treatment with sertraline 50 mg can benefit from an additional 4-week treatment with sertraline 100 mg and that the PEDT may be a valid measure of treatment response in PE patients.

Effect of zinc source on performance, zinc status, immune response, and rumen fermentation of lactating cows.

Two experiments were conducted to examine the effect of zinc (Zn) source on the performance, Zn status, immune response, and rumen fermentation of lactating cows to find the most available Zn source for dairy production. In Experiment 1, a total of 30 multiparous Holstein cows were randomly allocated by body weight and milk yield to one of five treatments in a completely randomized design. Cows were fed a total mixed ration (TMR) with no Zn addition (containing 37.60 mg Zn/kg TMR by analysis), and the basal TMR supplemented with 40 mg Zn/kg TMR from either Zn sulfate or one of three organic Zn chelates with weak (Zn-AA W), moderate (Zn-Pro M), or strong (Zn-Pro S) chelation strengths, respectively for 55 days. In Experiment 2, the in vitro rumen fermentation method was used in a completely randomized design involving a 4 × 3 factorial arrangement of treatments. The four Zn sources were the same as those used in Experiment 1, and the three supplemental Zn levels in the rumen fluid were 0, 10, and 20 µg/mL, respectively. The feed intake, milk composition, and somatic cell count (SCC) were unaffected (P > 0.05) by treatments. However, the milk yield was increased (P < 0.05) by addition of Zn from both the Zn-AA W and Zn-Pro S. Plasma Zn level at the end of the experiment was increased (P < 0.05) by addition of Zn from all three organic sources. Serum antibody titers on day 21 after vaccination with foot and mouth disease (FMD) vaccine were increased (P < 0.05) by both supplemental Zn-AA W and Zn-Pro S. The organic Zn sources with different chelation strengths supplemented at the added Zn level of 10 µg/mL were more effective (P < 0.05) in improving the rumen fermentation than Zn sulfate, with the most effective being Zn-AA W. In conclusion, Zn source had no influence on the feed intake, milk composition, and SCC; however, both the Zn-AA W and Zn-Pro S were more effective than Zn-Pro M and Zn sulfate in enhancing the rumen fermentation, Zn status, and humoral immune response as well as improving milk yield of lactating cows. The improved milk production might be attributed to the improved rumen fermentation, Zn status, and immune function.

Nutritional value of dried fermentation biomass, hydrolyzed porcine intestinal mucosa products, and fish meal fed to weanling pigs.

Dried fermentation biomass (DFB) and hydrolyzed porcine intestinal mucosa are co-products of L-Lys • HCl production and heparin extraction, respectively. Three experiments were conducted to determine standardized ileal digestibility (SID) of AA (Exp. 1), concentration of DE and ME (Exp. 2), and standardized total tract digestibility (STTD) of P (Exp. 3) in DFB and 2 hydrolyzed porcine intestinal mucosa products (PEP50 and PEP2+), and compare these values with values for fish meal. In Exp. 1, 12 ileal cannulated barrows (BW = 11.5 ± 1.1 kg) were allotted to a replicated 6 × 6 Latin square design with 6 diets and 6 periods. A N-free diet, diet based on soybean meal (SBM), and 4 diets based on a combination of SBM and DFB, PEP50, PEP2+, or fish meal were formulated. With the exception of Lys, there were no differences in SID of indispensable AA between DFB and fish meal. Except for Thr, no differences in SID of indispensable AA between PEP50 and fish meal were observed, but SID of all indispensable AA, except Lys and Trp, was less (P < 0.05) in PEP2+ than in the other ingredients. In Exp. 2, 40 barrows (BW = 12.8 ± 1.4 kg) were allotted to 5 diets with 8 pigs/diet. A basal diet containing 96.4% corn and 4 diets containing corn and DFB, PEP50, PEP2+, or fish meal were formulated. The DE (5,445 kcal/kg DM) and ME (5,236 kcal/kg DM) in DFB were greater (P < 0.01) than in PEP50 (4,758 and 4,512 kcal/kg DM for DE and ME, respectively) and fish meal (4,227 and 3,960 kcal/kg DM for DE and ME, respectively). Also, DE in DFB was greater (P < 0.01) than in PEP2+ (4,935 kcal/kg DM), but ME in DFB was not different from that in PEP2+ (4,617 kcal/kg DM). Furthermore, DE in PEP50 and PEP2+ were greater (P < 0.01) than in fish meal, but ME did not differ from that in fish meal. In Exp. 3, 40 barrows (BW = 12.4 ± 1.3 kg) were randomly allotted to 5 diets with 8 pigs/diet. A P-free diet and 4 diets in which the sole source of P was from DFB, PEP50, PEP2+, or fish meal were formulated. The STTD of P in DFB (96.9%) and PEP2+ (97.6%) were greater (P < 0.01) than in PEP50 and fish meal (76.2% and 68.5%, respectively), and STTD of P in PEP50 was greater (P < 0.01) than in fish meal. In summary, SID of most indispensable AA did not differ among DFB, PEP50, and fish meal, but DE and ME and STTD of P in DFB were greater than in PEP50 and fish meal.

Amperometric nitric oxide sensor based on nanoporous platinum phthalocyanine modified electrodes.

This article describes the fabrication of electropolymerized Metallo 4', 4″, 4‴, 4'''' tetra-amine phthalocyanine (poly-MTAPc) modified electrodes for the detection of nitric oxide (NO) in phosphate-buffered saline (PBS) at pH 7.4. A two-step synthetic protocol using a laboratory microwave reactor was adopted to provide three MTAPc complexes bearing different metal centers (M = Cu(2+): CuTAPc, M = Zn(2+): ZnTAPc, and M = Pt(2+): PtTAPc). The MTAPc complexes and the intermediates were characterized by MALDI-TOF mass spectrometry, UV-vis spectroscopy, (1)H NMR spectroscopy, and elemental analysis. The MTAPc products were separately electropolymerized either onto a glassy carbon (GC) electrode as a thin-film or within the pores of Anodisc nanoporous alumina membrane as a densely packed array of poly-MTAPc nanotubes to produce two electrode systems. In the latter system, the surface area enhancement provided by the nanotube-arrayed morphology of the poly-MTAPc enabled a high faradaic (signal) to capacitative (background) current during NO electro-oxidation. Amperometric detection of NO using these two electrode systems shows that the sensitivity and linear ranges were insensitive to the metal centers (M = Cu(2+), Zn(2+), and Pt(2+)) of the poly-MTAPc material.

A role for trichosanthin in the expansion of CD4CD25 regulatory T cells.

CD4(+)CD25(+) regulatory T cells (Tregs) are critical for the peripheral immune tolerance. Understanding the signals for the generation of Tregs is important for the clinical immunotherapy, but only limited progress has been made on obtaining enough peripheral Tregs. The aim of this study was to evaluate the role of trichosanthin (Tk) extracted from Chinese medicinal herb Trichosanthes kirilowi on the function of Tregs in vitro and in vivo. We reported here that Tk is needed for the expansion of freshly isolated CD4(+)CD25(+)Tregs (nTregs) into Tk-expanded CD4(+)CD25(+)Tregs (Tk-Tregs) through up-regulating CD25 and Foxp3 expression. The dose-response analyses indicated that 100 ng/ml Tk was the most appropriate dose. The result of real-time PCR showed that Tk-Tregs expressed 1.5-fold higher levels of Foxp3 than those observed in nTregs. Tk-Tregs markedly suppressed activation of effector T cells at a suppressor/responder ratio of 1:1, 1:2, 1:4, 1:8 or 1:16, and their effect was dose dependent. Moreover, Tk-Tregs secreted more immunosuppressive cytokines interleukin (IL)-10 and transforming growth factor (TGF)-beta1 after stimulating with antigen and antigen-presenting cells (APC). Transwell experiments showed that not only cell-to-cell contact but also soluble cytokines were involved in suppressive mechanism of Tk-Tregs. And Tk-Tregs were more efficient in suppressing CD25(-)T cell response to specific antigen than to irrelative antigen. Most importantly, it was revealed for the first time that Tk-Tregs could prolong the survival duration of mice with acute graft-versus-host disease (aGVHD). In conclusion, the study suggests a possible therapeutic potential of Tk-Tregs for clinical treatment on aGVHD.

Catalpol attenuates MPTP induced neuronal degeneration of nigral-striatal dopaminergic pathway in mice through elevating glial cell derived neurotrophic factor in striatum.

The protective effect of an iridoid catalpol extracted and purified from the traditional Chinese medicinal herb Rehmannia glutinosa on the neuronal degeneration of nigral-striatal dopaminergic pathway was studied in a chronic 1-methyl-4-phenyl-1,2,3,4-tetrahydropyridine (MPTP)/probenecid C57BL/6 mouse model and in 1-methyl-4-phenylpyridimium (MPP(+)) intoxicated cultured mesencephalic neurons. Rotarod performance revealed that the locomotor ability of mice was significantly impaired after completion of model production and maintained thereafter for at least 4 weeks. Catalpol orally administered for 8 weeks (starting from the second week of model production) dose dependently improved the locomotor ability. HPLC revealed that catalpol significantly elevated striatal dopamine levels without changing the metabolite/dopamine ratios. Nor did it bind to dopamine receptors. Therefore it is unlikely that catalpol resembles any of the known compounds for treating Parkinsonism. Instead, catalpol dose dependently raised the tyrosine hydroxylase (TH) neuron number in substantia nigra pars compacta (SNpc), the striatal dopamine transporter (DAT) density and the striatal glial cell derived neurotrophic factor (GDNF) protein level. Linear regression revealed that both the TH neuron number and DAT density were positively correlated to the GDNF level. In the cultured mesencephalic neurons, MPP(+) decreased the dopaminergic neuron number and shortened the neurite length, whereas catalpol showed protective effect dose dependently. Furthermore, the expression of GDNF mRNA was up-regulated by catalpol to a peak nearly double of normal control in neurons intoxicated with MPP(+) for 24 h but not in normal neurons. The GDNF receptor tyrosine kinase RET inhibitor 4-amino-5-(4-methyphenyl)-7-(t-butyl)-pyrazolo-[3,4-d]pyrimidine (PP1) abolished the protective effect of catalpol either partially (TH positive neuron number) or completely (neurite length). Taken together, catalpol improves locomotor ability by attenuating the neuronal degeneration of nigral-striatal dopaminergic pathway, and this attenuation is at least partially through elevating the striatal GDNF expression.

De novo urothelial carcinoma in kidney transplant patients with end-stage aristolochic acid nephropathy in China.

OBJECTIVE: Aristolochic acid nephropathy (AAN) is a progressive renal interstitial fibrosis disease that was initially reported among a Belgian cohort of about 50 patients after the intake of diet pills containing the Chinese herb Aristolochia fangchi. In addition to renal disease, foci of AAN show increased incidences of urothelial carcinomas (UC). Immunosuppression is associated with an increased risk for the development of different malignancies. Our aim was to examine the outcomes of UC among patients with AAN after transplantation in China, the cradle of this traditional medicine. PATIENTS AND METHODS: We performed a retrospective evaluation of the charts and pathology reports of 1612 renal transplant recipients treated at our 2 institutions. RESULTS: From January 1998 to December 2006, we performed cadaveric kidney transplantations in 17 patients with AAN, all of whom were treated with cyclosporine plus azathioprine or mycophenolate mofetil plus prednisone. One-year graft survival was 100%. During the mean follow-up of 57 months (range, 21-108 months), 9 recipients (52.9%) developed UC, compared with a 0.46% prevalence of urinary tract tumors among other Chinese kidney transplant recipients. The age at which the diagnosis was made ranged from 39 to 66 years (mean, 53.6 +/- 6.8 years). Among the 9 patients with UC, 8 cases (88.9%) involved the upper urinary tract: bilateral, 3 cases, 37.5%; unilateral, 5 cases, 62.5%. In 1 patient only a bladder tumor was detected. Two patients showed the bladder, synchronous bilateral ureter, and pelvis to be involved. All patients with UC underwent surgical treatment, recovering uneventfully with functioning grafts after tumor excision, except 1 patient who underwent nephrectomy of the transplanted kidney. Six patients (75%) experienced recurrences during the follow-up period. Three patients died within a mean of 20 months (range, 1-42 months) after tumor excision. CONCLUSIONS: The risk for UC is increased among patients with AAN after transplantation. Regular screening for early detection of malignancy is mandatory. Longer follow-up and results from other transplant centers are needed to further investigate the relationship between AAN and UC among renal transplant patients.