OBJECTIVE:To establish UV-spectrophotometry method for the determination of melatonin in the compound melatonin capsules.METHODS:The UV-spectrophotometry was adopted with the detection wavelength at277nm.RESUL_ TS:There was a good linear relationship between melatonin and its absorbability(r=0.9996)within the concentration range of5～25?g/ml.The average recovery was96.49%(RSD=1.21%,n=6).CONCLUSION:The method is accurate,reliable,convenient and stable.

Objective:To explore clinical therapeutic effect and safety of diltiazem treating coronary artery spasm. Methods:A total of 100 patients with coronary artery spasm were selected in our hospital.According to number ta-ble method,they were randomly and equally divided into routine treatment group (n=50,received,lipid regulation and expanding coronory artery etc.)and diltiazem group (n=50,received diltiazem pumping based on routine treat-ment).Clinical and ECG therapeutic effect,changes of blood lipids,nitric oxide (NO)and endothelin-1 (ET-1) levels before and after treatment and safety were compared between two groups.Results:(1 )Compared with rou-tine treatment group,there were significant increase in total effective rates of clinical effect (60.00% vs.94.00%) and ECG (62.00% vs.92.00%)in diltiazem group,P<0.01 both;(2)Compared with before treatment,there were significant improvements in all ECG indexes,heart rate and blood pressure in two groups after treatment,and those of diltiazem group were significantly better than those of routine treatment group (P<0.05 all);(3)Com-pared with before treatment and routine treatment group,there were significant improvements in blood lipid levels in diltiazem group after treatment (P<0.05 all);(4)Compared with before treatment and routine treatment group,there was significant rise in NO level [(95.17±19.99)μmol/L,(95.17±21.22)μmol/L vs.(106.71± 22.38)μmol/L]and significant reduction in ET-1 level [(64.09±16.29)ng/L,(59.98±14.28)ng/L vs.(44.91± 6.38)ng/L]in diltiazem group after treatment (P<0.05 all);(6)During treatment,there were no adverse reac-tion in both groups.Conclusion:Diltiazem based on routine medication possess significant therapeutic effect and high safety for treating coronary artery spasm,which is worth extension in clinic.

Objective To report clinical symptoms of a Chinese pedigree of familial paramyotonia congenital (PMC) with progressive myopathy (PM), and investigate the mutations of hot spots in the adult skeletal muscle sodium channel α-subunit (SCN4A). Methods The medical history and clinical phenotype of the patients from this large family with PMC were collected. Insertional and spontaneous activity were recorded by routine electromyograph (EMG), and the exercise test (ET) and cool water test were also performed on some patients during episodes. The mutations of SCN4A were screened by PCR-SSCP and DNA sequencing in affected and unaffected members. Results The family is a four-generation kindred with 15 members affected by severe, homogeneous paralysis periodiea paramyotoniea pheuotype. The onset was early, and almost all patients developed severe progressive myopathy by middle age. Routine EMG shows myotonia discharge in all affected subjects. The compound remarkably motor action potential (CMAP) decreased more than 40% after ET with greater decreases in cool water test than in ET. The mutation screening study revealed a missense mutation (Met1592Val) in SCN4A in patients. Conclusions Autosomal dominant inheritance pattern with complete penetrance was observed in this family. The phenotype is in accord with that reported in other ethnic populations with more severe symptoms. The ET and cool water tests may be used as an easy and reliable diagnostic method. Our research supports that periodic paralysis and paramyotonia can be caused by the same mutation in SCN4A. Mutation Met1592Val is a hotspot for mutation screening in patients with PMC accompanied by PM in the Chinese population.

Objective To investigate the differential expression levels and significance of regulatory T cell (Treg) and T helper 17 (Th17) related immunologic factors in peripheral blood of arsenic-exposed rats.Methods Thirty-two Wistar rats were numbered by weight,randomly divided into four groups [control,low (1.25 ml/kg),medium (2.50 ml/kg),and high (5.00 ml/kg)],8 rats per group.Rats in control group were given oral gavage of deionized water,and low,medium and high arsenic exposed groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week.After 4 months,the urine and peripheral blood samples of rats were collected,urinary arsenic (uAs) was detected by inductively coupled plasma-mass spectrometry (ICP-MS),the results were shown in [median (minimum and maximum)],uAs was corrected by urinary creatinine (uCr),the unit was μg/g Cr;enzyme-linked immune-sorbent assay (ELISA) was applied to detect the levels of Treg,Th17,T lymphocytes activation related immunologic factors [interleukin-10 (IL-10),transforming growth factor beta1 (TGF-31),IL-17,IL-6,IL-2],the results were shown in (x) ± s.Results The uAs of the rats was compared between control,low,medium,and high arsenic exposed groups [7.50 (3.16-9.81),72.34 (62.34-106.63),209.15 (154.41-232.20),369.73 (289.50-516.55) μg/g Cr],the differences were statistically significant (F =337.55,P ＜ 0.05).IL-10 [(85.03 ± 7.11),(93.96 ± 8.14),(97.48 ± 6.23),(93.47 ± 4.41) ng/L],TGF-β1 [(72.88 ± 2.96),(81.45 ± 8.15),(86.08 ± 7.55),(90.29 ± 5.35) ng/L],IL-17 [(18.15 ± 3.66),(25.54 ± 5.59),(31.48 ± 5.74),(37.25 ± 7.36) ng/L],IL-6 [(83.68 ± 8.48),(85.14 ± 7.11),(89.78 ± 5.36),(93.48 ± 5.77) ng/L],and IL-2 [(80.65 ± 6.90),(73.86 ± 8.00),(69.93 ± 7.77),(62.06 ± 9.82) ng/L] of the rats were compared between control,low,medium,and high arsenic exposed groups,the differences were statistically significant (F =5.094,11.054,16.249,3.474,5.119,all P ＜ 0.05).There were positive correlations between uAs and TGF-β1,IL-17 concentration (r =0.723,0.605,all P ＜ 0.01),while IL-2 showed a negative correlation (r =-0.484,P ＜ 0.05).Concltsion Arsenic exposure could affect the secretion of Treg and Th17 related immunologic factors,so as to the imbalance of anti-inflammatory and pro-inflammatory,which may play a role in the formation and development of arsenic-related immune injury.

rse effect when compared with tibolone.

Objective:To understand the relationship between the disease condition of patients with coal-burning-borne endemic arsenic poisoning (abbreviated as coal-burning-borne arsenic poisoning) and serum lipid metabolism indicators.Methods:Using a case-control study method, in the coal-burning-borne arsenic poisoning village of Yuzhang Town, Qianxinan Prefecture, Guizhou Province, 204 patients with arsenic poisoning diagnosed according to the standard of "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) were included in case group, including 87 males and 117 females, aged(53.37 ± 8.06) years old; and they were divided into mild arsenic poisoning group (71 cases), moderate arsenic poisoning group (59 cases) and severe arsenic poisoning group (74 cases) according to the clinical grading. Another 63 residents were selected into control group in a non-arsenic-exposed village about 12 km away from the diseased village, including 23 males and 40 females, aged (53.78 ± 9.10) years old. A face-to-face questionnaire survey was conducted for each group of people, including basic information such as general demographic characteristics, smoking status, and drinking status; fasting peripheral blood was collected, and an automatic biochemical analyzer was used to detect serum total cholesterol (TC) and triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) levels.Results:There were significant differences of serum TC [(4.94 ± 1.00), (5.00 ± 0.99), (5.27 ± 0.94), (5.57 ± 1.07) mmol/L], TG [(2.17 ± 0.90), (2.25 ± 1.31), (2.66 ± 1.43), (2.78 ± 1.40) mmol/L], LDL-C [(2.51 ± 0.79), (2.74 ± 0.64), (2.97 ± 0.66), (3.15 ± 0.80) mmol/L], and HDL-C levels [(1.57 ± 0.55), (1.42 ± 0.43), (1.36 ± 0.42), (1.30 ± 0.38) mmol/L] in control group, mild, moderate and severe arsenic poisoning groups ( F = 5.83, 3.64, 9.72, 4.41, P < 0.01 or < 0.05). Among them, the serum TC level in severe arsenic poisoning group, serum TG and LDL-C levels in moderate and severe arsenic poisoning groups were significantly higher than those in control group ( P < 0.05); the serum HDL-C level in moderate and severe arsenic poisoning groups were lower than that in control group ( P < 0.05); the serum TC, TG and LDL-C levels in severe arsenic poisoning group were significantly higher than those in mild arsenic poisoning group ( P < 0.05). After linear trend test, serum TC, TG and LDL-C levels all showed an upward trend with the degree of arsenic poisoning ( Ftrend = 15.77, 10.14, 29.15, P < 0.05), and serum HDL-C level showed a downward trend with the degree of arsenic poisoning ( Ftrend = 12.75, P < 0.05). There were significant differences in the abnormal rates of serum TC, TG and LDL-C levels among control group and mild, moderate and severe arsenic poisoning groups (χ 2 = 21.16, 16.60, 8.29, P < 0.01 or < 0.05). Among them, the serum TC and TG levels abnormal rates in moderate and severe arsenic poisoning groups and serum LDL-C level abnormal rate in severe arsenic poisoning group were higher than those in control group ( P < 0.05), the serum TC, TG and LDL-C levels abnormal rates in severe arsenic poisoning group were higher than those in mild arsenic poisoning group ( P < 0.05). There was no significant difference of the serum HDL-C level abnormal rate among four groups (χ 2 = 2.11 , P > 0.05). The results of trend chisquare analysis showed that the abnormal rates of serum TC, TG and LDL-C levels presented an increasing trend with the degree of arsenic poisoning (χ 2trend = 19.90, 15.25, 7.63, P<0.05). The results of logistic regression analysis showed that the risk of abnormal serum TC level in patients with severe arsenic poisoning was 2.90 times that in control group [odds ratio ( OR) = 2.90, 95% confidence interval ( CI): 1.43 - 5.91], and the risk of abnormal serum LDL-C level in patients with severe arsenic poisoning was 2.87 times that in control group ( OR = 2.87, 95% CI: 1.22 - 6.71). Conclusion:There is a correlation between the disease condition of patients with coal-burning-borne arsenic poisoning and their dyslipidemia.

Objective To assess the expression pattern of protease-activated receptor 2 (PAR2) in human keratinocytes and to characterize its biological functions in the regulation of skin barrier.Methods Primary human keratinocytes and human N/TERT keratinocytes were used as the subject of this study.The expression and distribution of PAR2 in the keratinocytes were analyzed by using immunoflorescence staining and Western blot.Two different PAR2 agonists,trypsin and a PAR2-activating peptide (AP),as well as a PAR2-antagonistic peptide (H2N-FSLLRY-COOH) and a control peptide were used to induce the activation of PAR2 in the keratinocytes.Then,a fluorescence-based calcium mobilization assay was performed to evaluate the biological function of PAR2.Data were statistically analyzed by one-factor analysis of variance.Results Under normal culture conditions,PAR2 was weakly expressed in keratinocytes,and the expression was unaffected by culture medium composition or culture duration.Calcium mobilization was induced by trypsin of 50-250 nmol/L and the PAR2-activating peptide in a dose-and time-dependent pattern.The maximal activation of PAR2 was observed in keratinocytes treated with the PAR2 agonist HAN-SLIGKV-COOH of 75-250 μmol/L.The PAR2-antagonistic peptide (H2N-FSLLRY-COOH) obviously suppressed the increase in calcium mobilization induced by trypsin,while the control peptide PAR-RAP showed no inductive effect on the PAR2 activation based on the absence of calcium mobilization.The substrate-induced calcium release was complete within 250 seconds,and peaked at 50 seconds after the initial trypsin or PAR-AP stimulation.Moreover,the activation of PAR2 was accompanied by an increase in ERK phosphorylation and elicitation of MAPK signaling pathway in keratinocytes.Conclusions Human keratinocytes positively express PAR2,which can be activated by trypsin and PAR2-activating peptides,and the activation of PAR2 may influence the physiological function of keratinocytes by inducing intracellular calcium release.

The corona virus disease 2019 (COVID-19) has recently become pandemic and is still spreading. Many severe or critical COVID-19 cases meet the diagnostic criteria of sepsis and septic shock in the clinical manifestations. It is important to study the pathogenesis and treatment strategy of COVID-19 for the disease prevention and control. This article reviews the clinical features and treatment progress of COVID-19 viral sepsis.

OBJECTIVETo investigate the effects of two different dosages of conjugated equine estrogen (CEE) on preventing bone loss and relieving the symptoms of menopausal syndrome in women at an early stage of menopause.

METHODSA total of 236 postmenopausal women were randomly allocated to one of the following groups: Group A: 0.625 mg CEE + 2 mg medroxyprogesterone acetate (MPA) + 1 tab Caltrate-D per day; Group B: 0.3 mg CEE + 2 mg MPA + 1 tab Caltrate-D per day; Group C: 1 tab Caltrate-D per day as the control group. The study was continued for 2 years.The following parameters were monitored: (1) L2-4 bone mineral density (BMD) (measured with dual energy X-ray absorptiometry (DEX)), (2) menopausal syndrome improvement (assessed by comparing Kupperman scores), (3) vaginal bleeding rate, and the thickness of the endometrium and breast in each group.

RESULTSOverall, 213 cases (90%) completed the 1-year study and 176 cases (75%) completed the 2-year study. The percentage changes in L2-4 BMD at the 12th and 24th month in Group A were +2.3% and +3.7%, respectively, with the posttreatment values being significantly higher than pretreatment values (P < 0.001). The percentage changes were +2.7% at 12th month (P < 0.05) and +0.7% at 24th month (P > 0.05) in Group B. And that of Group C were -0.4% at 12th month and -1.6% at 24th month (P > 0.05). L2-4 BMD in both Group A and B was significantly higher than that in Group C at 12th and 24th month (A vs C, P < 0.001; B vs C, P < 0.05). Kupperman Scores were significantly reduced after 1, 3, 6, 12 and 24 months in all 3 groups when compared with baseline (P < 0.001). Scores in Group A and Group B were significantly lower than that in Group C (P < 0.001). However, the vaginal bleeding rates in Group A were significantly higher than that in Group B or in Group C. There was no atypical hyperplasia of endometrium in the 3 groups by the end of the study. One patient in Group A developed superficial thrombophlebitis by the end of 12th month.

CONCLUSIONContinuous combination of CEE and MPA is effective in preventing bone loss and relieving the symptoms of menopausal syndrome in women at an early stage of menopause. The vaginal bleeding rates in the Group treated with 0.625 mg/d CEE were significantly higher than those treated with 0.3 mg/d CEE.

Alkaline Phosphatase ; blood ; Bone Density ; drug effects ; Endometrium ; pathology ; Estrogens, Conjugated (USP) ; administration & dosage ; Female ; Hormone Replacement Therapy ; adverse effects ; methods ; Humans ; Menopause ; Middle Aged ; Progestins ; administration & dosage ; Uterine Hemorrhage ; epidemiology

Objective: To explore the effects of sodium arsenite (NaAsO₂) exposure on the activation and extracellular matrix secretion of human hepatic stellate cells, and to provide a theoretical basis for the mechanism study of arsenic induced hepatic fibrosis. Methods: Different doses of NaAsO₂ (0.0, 0.1, 1.0, 10.0, 50.0, 100.0 μmol/L) were exposed to human hepatic stellate cell line (Lx-2) for 24, 48 and 72 huors. CCK-8 assay was used to measure cell viability and IC₅₀ of NaAsO₂ on Lx-2 was then calculated; According to IC₅₀ results, 0.000, 1.875, 3.750, 7.500, and 15.000 μmol/L of NaAsO₂ were exposed to Lx-2 cells for 24 hours, besides, 7.500 μmol/L of NaAsO₂ was exposed to Lx-2 cells for 0, 12, 24, 48, and 72 hours, then collected cells and culture supernatant; HSC activation-related protein, including α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1) expression levels were detected by Western blot analysis, the main extracellular matrix including laminin (LN) , hyaluronic acid (HA), collagen Ⅳ (COL-Ⅳ) and procollagen Ⅲ(P Ⅲ NP) secretion level was detected by Elisa assay. Results: CCK-8 assay showed that the cell viability of Lx-2 cells were increased obviously at low doses (≤1.0 μmol/L) of arsenic exposure, especially at 48 and 72 h. In contrast, with the increasing doses of arsenic exposure, the survival rate of Lx-2 cell was decreased gradually, and the survival rate of the high-dose (50, 100 μmol/L) arsenic exposure group at 24, 48 and 72 h were significantly lower than 0.0 μmol/L group, P<0.05. The IC₅₀ of NaAsO₂ on Lx-2 cells at 24, 48, 72 h were calculated as 72.75, 48.19 and 29.95 μmol/L, respectively; The expression levels of HSC activation-related protein showed that, after treated with 1.875, 3.750, 7.500, 15.000 μmol/L NaAsO₂ for 24 h, α-SMA and TGF-β1 protein level were higher than 0.000 μmol/L group. The increased expression of α-SMA and TGF-β1 protein were most significant in 7.500 μmol/L NaAsO₂ group (P<0.05). In addition, the expression levels of α-SMA and TGF-β1 also showed a time-dependent increasing in Lx-2 cells after treated with 7.500 μmol/L NaAsO₂ for 0, 12, 24, 48 and 72 h; Elisa assay showed that after treated with 1.875, 3.750, 7.500, 15.000 μmol/L NaAsO₂ for 24 h, the secretion levels of HA, LN, COL-Ⅳ and PⅢNP were obvious higher than 0.000 μmol/L group (P<0.05). Moreover, the secretion levels of HA, LN, COL-Ⅳ and P Ⅲ NP also showed a time-dependent increased manner in Lx-2 cells after exposed to 7.500 μmol/L NaAsO₂ for 0, 12, 24, 48 and 72 h (P<0.05). Conclusion NaAsO₂ exposure to Lx-2 cells can upregulate the expression level of HSC activation-related proteins, induce its further activation, then increase ECM secretion level.