Regulation of ischemic stroke by circadian rhythm and intervention by traditional Chinese medicine / 中国中药杂志

Circadian rhythm is an internal regulatory mechanism formed in organisms in response to the circadian periodicity in the environment, which modulates the pathophysiological events, occurrence and development of diseases, and the response to treatment in mammals. It significantly influences the susceptibility, injury, and recovery of ischemic stroke, and the response to therapy. Accumulating evidence indicates that circadian rhythms not only regulate the important physiological factors of ischemic stroke events, such as blood pressure and coagulation-fibrinolysis system, but also participate in the immuno-inflammatory reaction mediated by glial cells and peripheral immune cells after ischemic injury and the regulation of neurovascular unit(NVU). This article aims to link molecular, cellular, and physiological pathways in circadian biology to the clinical consequences of ischemic stroke and to illustrate the impact of circadian rhythms on ischemic stroke pathogenesis, the regulation of NVU, and the immuno-inflammatory responses. The regulation of circadian rhythm by traditional Chinese medicine is reviewed, and the research progress of traditional Chinese medicine intervention in circadian rhythm is summarized to provide a reasonable and valuable reference for the follow-up traditional Chinese medicine research and molecular mechanism research of circadian rhythm.

Identification of pathogenic genes in a family with oculocutaneous albinism / 中华皮肤科杂志

Objective:To detect mutations in a pedigree containing two brothers with oculocutaneous albinism (OCA) by whole-exome sequencing and Sanger sequencing.Methods:Clinical data were collected from a pedigree with OCA, and DNA was extracted from peripheral blood samples obtained from the proband and other family members. The whole-exome coding region of the proband was directly sequenced by whole-exome sequencing technology to identify potential pathogenic mutations, and Sanger sequencing was conducted to verify the gene mutations.Results:Both the proband and his younger brother presented with generalized white skin, golden-yellow hair, bilateral nystagmus, photophobia, translucent iris, conjunctival congestion, and refractive errors of both eyes. The proband′s parents, grandparents, maternal grandparents, and children were all phenotypically normal, and his parents′ marriage was non-consanguineous. Three heterozygous mutations were identified in the OCA2 gene of both the proband and his younger brother, including a nonsense mutation c.1290T>A, and 2 missense mutations c.1363A>G and c.1204T>C. The mutation c.1204T>C has not been previously reported, and was a novel gene mutation in the OCA2 gene. In addition, 1 heterozygous mutation c.1204T>C was identified in the OCA2 gene in the proband′s father and daughter, 2 heterozygous mutations c.1290T>A and c.1363A>G were found in the OCA2 gene in the proband′s mother, and 1 heterozygous mutation c.1290T>A was identified in the OCA2 gene in the proband′s son and the daughter of the proband′s younger brother.Conclusions:Three gene mutations were identified in the OCA2 gene in the 2 patients with OCA, and the nonsense mutation c.1290T>A may be the pathogenic mutation causing the clinical phenotype of this family. These findings expand the pathogenic mutational spectrum of the OCA gene.

Detection of Chlamydia trachomatis DNA in urine and urethral/cervical swab samples: analysis of results from 1 475 outpatients attending sexually transmitted disease clinics / 中华皮肤科杂志

Objective:To compare the detection rate of genital Chlamydia trachomatis (CT) DNA between urine and urethral/cervical swab samples. Methods:From December 2018 to December 2019, a total of 1 475 outpatients were collected from sexually transmitted disease clinics in 7 medical institutions, such as Department of Venereology, Guangzhou Institute of Dermatology, including 1 118 males and 357 females. One urethral/cervical swab sample and one urine sample were collected successively from each patient. Real-time fluorescence-based PCR was performed to detect CT DNA in urine and urethral/cervical swab samples, and paired chi-square test was used to compare the positive rate of CT DNA between the 2 kinds of samples. Random- or fixed-effect meta-analysis was conducted for the test of heterogeneity and merging of positive rates of CT DNA in the urine and urethral/cervical swabs among 7 medical institutions.Results:The positive rate of CT DNA in the urine samples was significantly higher than that in the swab samples from 4 medical institutions (all P < 0.05) , while there was no significant difference in the positive rate of CT DNA between the 2 kinds of samples from 3 medical institutions (all P > 0.05) . The heterogeneity ( I2) estimates of the CT-DNA positive rate in urine and swab samples among different medical institutions were 78.6% (95% CI: 55.9% - 89.6%) and 73.7% (95% CI: 43.7% - 87.7%) , respectively; meta-analysis showed that the total merged positive rate of CT DNA in the urine samples was 10.8% (95% CI: 7.2% - 15.9%) , which was significantly higher than that in the swab samples (7.8%, 95% CI: 4.9% - 12.1%; χ2 = 39.2, P < 0.05) . Compared with the swab sample-based CT-DNA detection method, the sensitivity, specificity, positive predictive value, negative predictive value and consistency rate of the urine sample-based CT-DNA detection method were 97.0% (128/132) , 96.3% (1 293/1 343) , 71.9% (128/178) , 99.7% (1 293/1 297) , and 96.3% (1 421/1 475) , respectively. The positive rate of CT DNA in the urine samples from 1 118 male patients was 11.0% (95% CI: 7.2% - 16.5%) , which was significantly higher than that in the swab samples (7.6%, 95% CI: 4.9% - 11.8%; χ2 = 34.3, P < 0.05) . There was no significant difference in the positive rate of CT DNA between the urine (11.9%, 95% CI: 7.7% - 17.9%) and cervical swab samples from 357 female patients (10.4%, 95% CI: 7.6% - 14.0%; χ2 = 3.2, P > 0.05) . Conclusions:The positive rate of CT DNA in urine samples is higher than or similar to that in urethral/cervical swab samples. The urine sample-based CT-DNA detection method has characteristics of convenience, non-invasiveness, painlessness and low cost, and is worthy of clinical promotion.

Acute and chronic toxicity of 0.5% podophyllotoxin-loaded nanostructured lipid carriers to vaginal mucosa in rabbits and rats / 南方医科大学学报

OBJECTIVE@#To test the acute and chronic toxicity of topical application of 0.5% podophyllotoxin-loaded nanostructured lipid carriers (POD-NLC) to the vaginal mucosa.@*METHODS@#Twelve New Zealand rabbits were randomized into 3 groups and subjected to daily topical applications of normal saline (control group), 0.5% podophyllotoxin tincture (POD-T) or 0.5% POD-NLC on the vaginal mucosa for 10 consecutive days, and the pathological changes in the mucosa were graded using the Eckstein scoring system.The acute toxicity of POD-NLC was tested in 20 SD female rats, which received intravaginal administration of POD-NLC or vehicle for 3 times within 24 h; After 14 days of continuous observation, the rats were dissected for calculating the viscera coefficient.For testing the chronic toxicity of POD-NLC, 80 SD female rats were randomized into 4 groups and subjected to daily intravaginal administration of the vehicle or POD-NLC at low, moderate or high doses for 13 consecutive weeks.The rats were weighed once a week and at the end of the experiment, 2/3 of the rats from each group were sacrificed to collect blood samples, calculate the viscera coefficient, and examine the pathological changes in the liver.The remaining 1/3 rats were observed for another 2 weeks without further drug treatment and the same examinations were performed.@*RESULTS@#In the rabbits, 0.5% POD-NLC elicited only mild irritation while POD-T caused moderate irritation of the vaginal mucosa.In the acute toxicity test, the organ coefficients were comparable between the rats treated with the vehicle and POD-NLC (>0.05).Long-term intravaginal administration of POD-NLC did not produce significant changes in the behavior, activity, body weight, blood biochemical profiles or organ coefficient as compared with the vehicle control group (>0.05).@*CONCLUSIONS@#Intravaginal administration of 0.5% POD-NLC causes very mild irritation without obvious acute or chronic toxicity to the vaginal mucosa in rabbits and rats.

Analysis of the clinical and genetic characteristics of a progressive symmetric erythrokeratoderma / 中国医师杂志

Objective To investigate the clinical and genetic characteristics of a progressive symmetric erythrokeratoderma (PSEK).Methods The clinical and genetic characteristics of a PSEK were compared and analyzed with clinical collection and review of 17 PSEK families reported in China since 1980.Results PSEK was consistent with autosomal dominant inheritance.PSEK had the feature with incomplete penetrance and variable expression.The clinical features included hyperkeratotic plaques with distinct border and strikingly symmetric distribution pattern on the extremities.Part of patients was extended to other areas of the body.The onset of the disease commonly started in infancy or childhood.The general health condition was not affected.PSEK might be associated with other clinical symptoms.The diseased potential increased in the family of consanguineous marriage.Conclusions PSEK has genetic heterogeneity.Its causative genes have not been determined.Further studies are needed.

Distribution of podophyllotoxin-loaded nanostructured lipid carriers after topical application on cervical mucosa in Tibet minipigs / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the distribution and systemic toxicity of podophyllotoxin-loaded nanostructured lipid carriers (POD-NLC) after topical application on the cervical mucosa in Tibet minipigs.</p><p><b>METHODS</b>Twelve Tibet mini-pigs were randomized into test group and control group to receive topical application of 0.5% POD-NLC and 0.5% POD tincture, respectively, on the cervical mucosa. Cervical mucosal irritation, targeted distribution and systemic absorption of POD were observed at different time points within 24 h after the drug application.</p><p><b>RESULTS</b>No local inflammation reaction was observed in the test group, while serious local irritations (swelling, blisters, blood blisters, erosion and ulceration) occurred in the control group. The fluorescence intensity of POD in the mucosal tissue reached the peak level at 4 h after drug application in the control group, while the POD fluorescence intensity increased slowly and reached the peak level at 16 h in the test group. The peak blood POD concentration occurred at 6 h after POD-NLC application in the test group (14.28∓0.33 ng/mL), as compared to 4 h in the control group (42.46∓0.32 ng/mL). At all the time points within 24 h, blood POD concentration remained significantly lower in the test group than in the control group (P<0.05), and the area under curve of blood POD concentration in the control group was 1.38-fold greater than that in the test group.</p><p><b>CONCLUSION</b>POD-NLC allows sustained release of POD and achieves a higher POD concentration in the mucosal tissue without causing local irritation or obvious systemic toxicity in Tibet minipigs.</p>

Mutation analysis of the COL7A1 gene in pretibial dominant dystrophic epidermolysis bullosa / 中华皮肤科杂志

Objective To explore the role of type Ⅶ collagen (COL7A1) gene in the pathogenesis of pretibial dominant dystrophic epidermolysis bullosa (DDEB-Pt).Methods Peripheral blood samples were obtained from a sporadic Chinese patient of Han nationality with DDEB-Pt,his parents and 100 healthy human controls.A modified salting-out method was used to extract genomic DNA from the blood samples,and PCR was performed to amplify 118 exons of the COL7A1 gene followed by DNA sequencing.Results A G→A mutation was identified at position 6109 (G6109A) in exon 78 of the COL7A1 gene in this patient,which caused a change from GCT to ACT at codon 2037 in the triple helix region,and resulted in the substitution of glycine (Gly) by arginine (Arg) (p.Gly2037Arg).Conclusion A novel glycine substitution mutation was identified in the COL7A1 gene in the patient with DDEB-Pt,which may be a pathogenic mutation.

Expression of BATF, a member of the activator protein-1 family, in renal tissues of MRL/lpr mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of BATF, a member of the activator protein-1 family, in the renal tissues of mice with lupus nephritis.</p><p><b>METHODS</b>The renal tissues from 24-week-old female MRL/lpr mice and age- and sex-matched C57BL/6 mice were examined for BATF protein expressions using Western blotting and for expressions of BATF, RORγt and IL-17A mRNA using quantitative real-time PCR. The results of laboratory examinations were analyzed in relation with the histopathology of the mice.</p><p><b>RESULTS</b>The urinary protein and ds-DNA levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). Compared to normal control mice, MRL/lpr mice showed obvious glomerular fibrosis and inflammatory cell infiltrating with significantly increased BATF protein and mRNA expressions (P<0.05) and RORγt and IL-17 mRNA expressions in the renal tissues (P<0.05). In MRL/lpr mice, the expression of BATF mRNA was positively correlated with the RORγt and IL-17A mRNA expressions in the renal tissues.</p><p><b>CONCLUSION</b>The renal expressions of BATF protein and mRNA is increased in MRL/lpr mice. BATF may participate in the immunopathogenesis of lupus nephritis by enhancing Thl7 cell response.</p>

Role of endoplasmic reticulum stress pathway in podophyllotoxin nanostructured lipid carriers-induced apoptosis of VK2/E6E7 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the mechanism of podophyllotoxin nanostructured lipid carriers (POD-NLC)-induced apoptosis of VK2/E6E7 cells mediated by endoplasmic reticulum stress (ERS).</p><p><b>METHODS</b>VK2/E6E7 cells cultured in vitro were exposed to 0.125, 0.25, and 0.5 µg/ml POD-NLC or blank NLC for 24 h. The intracellular calcium concentration was measured by laser scanning confocal microscopy (LSCM), and the expression levels of GRP78, GRP94, and calpain2 mRNA and proteins in the cells were detected using RT-PCR and Western blotting.</p><p><b>RESULTS</b>Compared with the control cells, the cells exposed to POD-NLC showed a concentration-dependent increase of intracellular calcium concentration (P<0.01), and the differences were statistically significant between different dose groups (P<0.05). RT-PCR and Western blotting showed that POD-NLC up-regulated GRP78, GRP94 and calpain2 mRNA and proteins expressions, which showed significant differences between blank-NLC and the control groups (P>0.05).</p><p><b>CONCLUSION</b>POD-NLC induces apoptosis of VK2/E6E7 cells possibly by triggering the endoplasmic reticulum stress response.</p>

Influence of traditional Chinese medicine constitution type on the susceptibility of hypertensive cases to intracerebral haemorrhage / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the influence of Chinese medicine constitution type on the susceptibility of hypertensive patients to intracerebral haemorrhage.</p><p><b>METHODS</b>Primary hypertensive patients were studied and divided into the hypertension and the hypertensive intracerebral haemorrhage groups, depending on whether or not the patients had intracerebral haemorrhage. The demographic characteristics, physiological characteristics, living habits, biochemical tests, other chronic diseases, Chinese medicine constitution type, etc. were collected and compared between the two groups. The neurological deficit in the hypertensive intracerebral haemorrhage group was also compared among the different constitution types.</p><p><b>RESULTS</b>A total of 304 patients participated in this investigation, including 213 cases in the hypertension group and 91 cases in the hypertensive intracerebral haemorrhage group. The percentages of dampness-heat and qi-depression types in the intracerebral haemorrhage group were greater than those with the same types in the hypertension group (20.9% vs 6.1%, 22.0% vs 8.0%; P<0.05). The percentages of each constitution type were different between genders in both groups. In addition, there were more male cases (14/20) with qi-depression type and more female cases (7/8) with phlegm-dampness type in the hypertensive intracerebral haemorrhage group than those with the same types (3/17 and 9/34, respectively) in the hypertension group (P<0.05). The cases with phlegm-dampness and qi-depression types had greater levels of blood lipids in the intracerebral haemorrhage group than those with the same types in the hypertension group (P<0.05). Furthermore, the cases with the qi-deficiency type presented with a more severe neurological deficit than those with the other types (P<0.05).</p><p><b>CONCLUSIONS</b>Chinese medicine constitution type might have an impact on the susceptibility of hypertensive patients to have an intracerebral haemorrhage. The heat-dampness and qi-depression types might lead to greater susceptibility than the other types. In addition, the patient's gender and blood lipids might also influence the susceptibility along with the constitution type.</p>

Study on the distribution of Chinese medical constitutions of hypertension complicated diabetes patients / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the distribution features of Chinese medical constitutions in hypertension complicated diabetes patients.</p><p><b>METHODS</b>Recruited were 251 primary hypertension inpatients at the Department of Neurology and the Department of Cardiology, Mindong Hospital of Ningde City from October 2010 to March 2011. They were assigned to two groups according to whether they were complicated with diabetes, i.e., the primary hypertension complicated diabetes (as the case group, 78 cases) and the primary hypertension without complicated diabetes (as the control group, 173 cases). The constitution types were investigated by questionnaire. The constitution type distribution was compared between the two groups. The data including gender, age, and the distribution of the constitution type were compared between the two groups. The levels of TG, TC, LDL-C, Hb, FPG, and ALB were detected on the 2nd day after admission. The levels of TG, TC, LDL-C, Hb, and ALB were compared be- tween the two groups in patients of yin deficiency constitution, phlegm dampness constitution, and qi deficiency constitution.</p><p><b>RESULTS</b>There was no statistical difference in the hypertension grading, the disease course, and chronic disease complications between the two groups (P > 0.05). The main constitution types were yin deficiency (accounting for 26.0%), phlegm dampness (accounting for 19.1%), and qi deficiency (accounting for 19.1%) in the control group. The main constitution types were yin deficiency (accounting for 32.1%), phlegm dampness (accounting for 30.8%), and qi deficiency (accounting for 17.9%) in the case group. The ratio of phlegm dampness type in the case group was higher than that in the control group with statistical difference (P = 0.041). There was no statistical difference in the constitution distribution in the same gender between the two groups (P > 0.05). There was no statistical difference in the constitution distribution in those younger than 80 years between the two groups (P > 0.05). Compared with those older than 80 years in the control group, the ratio of phlegm dampness was higher, and the ratios of yang deficiency, yin deficiency, qi deficiency, and dampness heat were lower in the case group with statistical difference (P = 0.020). There was no statistical difference in the constitution distribution among different age stages in the case group (P > 0. 05). But there was statistical difference in the constitution distribution among different age stages in the control group (P < 0.05). The yin deficiency and qi deficiency constitutions were dominated in thinner patients of the control group, while yin deficiency constitution was dominated in thinner patients of the case group, showing no statistical difference between the two groups (P > 0.05). There was no statistical difference in the distribution of constitution type in overweight patients between the two groups (P = 0.458). Compared with those of gentle type constitution in the same group, the levels of TC and LDL-C increased in those of phlegm dampness constitution in the two groups (P < 0.05). The level of TC increased in those of qi deficiency constitution in the case group. The level of Hb decreased in those of qi deficiency constitution in the control group (P < 0.05). Compared with those of qi deficiency constitution in the same group, the levels of TC and Hb obviously increased in those of phlegm dampness constitution in the control group (P < 0.05). The level of ALB increased in those of yin deficiency constitution in the case group (P < 0. 05). Compared with the control group, the level of FPG of those of each constitution increased in the case group (P < 0.05) ,.and the level of TC increased in those of qi deficiency constitution (P = 0.007).</p><p><b>CONCLUSIONS</b>The main constitution types of hypertension complicated diabetes patients were yin deficiency, phlegm dampness, and qi deficiency. The ratio of phlegm dampness was higher in hypertension complicated diabetes patients than hypertension without complicated diabetes patients. The levels of TC and LDL-C were higher in those of phlegm dampness constitution type. The level of TC was higher in hypertension complicated diabetes patients of qi deficiency constitution.</p>

Determination of eotaxin, tumor necrosis factor-α and interferon-γ levels in plasma from patients with chronic urticaria treated with the combination of BCG-PSN and cetirizine / 中华皮肤科杂志

Objective To investigate the changes to eotaxin,tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in plasma from patients with chronic urticaria treated with the combination of polysaccharide nucleic acid fraction of bacillus Calmette-Guerin (BCG-PSN) and cetirizine.Methods Totally,123 patients with chronic urticaria were enrolled into this study,and classified into two groups:the combination therapy group (n =60) treated with intramuscular BCG-PSN 2 ml every other day and oral cetirizine 10 mg once daily,and the monotherapy group (n =63) treated with oral cetirizine 10 mg once daily alone.The treatment lasted 54 days.Clinical efficacy was evaluated.Venous blood samples were collected from the patients before and after the treatment,as well as from 56 healthy controls.Enzyme-linked immunosorbent assay was performed to quantify the plasma levels of eotaxin,TNF-α and IFN-γ.Results At the end of the treatment,the total response rate was significantly higher in the combination therapy group than in the monotherapy group (88.3％ vs.63.4％,P ＜ 0.05).Before the treatment,no significant differences were observed in the plasma levels of eotaxin,TNF-α or IFN-γ between the two treatment groups,whereas the patients showed higher plasma levels of eotaxin and TNF-α but lower plasma level of IFN-γ compared with the healthy controls (all P ＜ 0.5).Both the combination therapy and monotherapy resulted in a statistical decrease in plasma eotaxin and TNF-α but an increase in plasma IFN-γ (all P ＞ 0.05),and the absolute values of changes in the three parameters were significantly higher in the combination therapy group than in the monotherapy group (eotaxin:(13.27 ± 4.11) μg/L vs.(8.12 ± 2.58) μg/L,t =8.3654,P ＜ 0.05; TNF-α:(12.38 ± 3.95) ng/L vs.(10.32 ± 3.41) ng/L,t =3.1005,P ＜ 0.05; IFN-γ:(17.06 ± 5.24) μg/L vs.(12.54 ± 4.07) μg/L,t =5.3573,P ＜ 0.05).Further more,the differences between the patients and healthy controls in the three parameters disappeared at the end of the treatment (all P ＞ 0.05).Conclusion BCG-PSN combined with cetirizine seems superior to cetirizine alone for the treatment of chronic urticaria.

Autologous peripheral blood hematopoietic stem cell transplantation in the treatment of three cases of severe pemphigus / 中华皮肤科杂志

ObjectiveTo explore the clinical efficacy and safety of autologous peripheral blood stem cell transplantation(APBSCT) in the treatment of pemphigus.MethodsTotally,3 patients with pemphigus vulgaris who responsed poorly to 6-month treatment with glucocorticoids or immunosuppressants or experienced aggrevation of disease and developed treatment-related complications,received APBSCT and were followed up for more than 5 years.There were 1 male and 2 females with an average age of 27.3(21-39) years.The mobilization program included cyclophosphamide (CTX) 4 g/m2,recombinant human granulocyte colonystimulating factors(G-CSF) and Rituximab 375 mg/m2,and the preconditioning regimen included intravenous CTX (50 mg/kg per day on days -6,-5,-4,-3),antithymocyte globulin at 2.5 mg/kg per day(on days -3,-2,-1 and 0) and Rituximab (600 mg/d on days 0 and 7).ResultsAll the 3.patients were successfully engrafted.The mean time for peripheral reconstruction:white blood cells 13.3 days (from day 11 to 16),platelet 16.3 days (from day 16 to 17).Monitoring of immunity indices and related antibodies showed no abnormality and the immune system was well reconstructed.No serious complications occurred during the follow up,and the patients' quality of life was obviously improved.ConclusionAPBSCT may be an effective and safe option for the treatment of pemphigus.

In vivo imaging of alopecia areata with confocal laser scanning microscopy / 中华皮肤科杂志

ObjectiveTo assess the microscopic features of alopecia areata(AA) by using in vivo confocal laser scanning microscopy(CLSM).MethodsBetween January 2010 and May 2011,26 male and 20 female patients diagnosed with AA were enrolled in this study.AA lesions and perilesional normal skin were examined by means of in vivo CLSM.Tissue specimens were obtained from the lesions and perilesional normal skin of 10 out of the 46 patients and subjected to routine histopathologic examination.ResultsThe number of hair follicles per unit area(cm2) was decreased in lesions of progressive AA and resting AA compared with normal perilesional skin(134.856 ± 18.301 and 147.159 ± 17.536 vs.301.613 ± 35.317,both P ＜ 0.05).Although the quantity of hair follicles increased in lesions of recovery AA((227.778 ± 16.861 )/cm2),but was still less than that in the normal perilesional skin (P ＜ 0.05).There was a lack of hair shaft in follicles,as well as an inflammatory infiltration in hair follicles,around hair follicles and capillaries in superficial dermis in lesions of progressive AA.The inflammatory infiltration was attenuated in lesions of resting AA.In lesions of recovery AA,the infiltration was further attenuated with an apparent growth of lanugos and terminal hairs.ConclusionsAs far as AA lesions are concerned,CLSM images are consistent with histopathological findings.CLSM may serve as a promising tool for monitoring efficacy and predicting prognosis based on hair growth cycle,hair follicle number,and inflammatory infiltration degree.

Preparation of podophyllotoxin nanostructured lipid carriers and its effects on immortalized human cervical epithelial cells with HPV infection in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of podophyllotoxin nanostructured lipid carriers (POD-NLC) on immortalized human cervical epithelial cells (H8) infected with HPV in vitro.</p><p><b>METHODS</b>POD-NLC was prepared by emulsion evaporation method and characterized using transmission electron microscopy, Zetasizer analyzer and high-performance liquid chromatography (HPLC). H8 cells were treated with different concentrations (0.0001-1 µg/ml) of POD-NLC, free POD, or blank nanostructured lipid carriers (NLC), and the cell proliferation was assessed using MTT assay to evaluate the cytotoxic effects. The changes of cell morphology were observed using fluorescence microscopy, and the cell cycle changes and cell apoptosis were analyzed using flow cytometry.</p><p><b>RESULTS</b>POD-NLC showed a spherical or elliptical shape with good stability in vitro. The average particle size of POD-NLC was 85.6∓10.25 nm, with a Zeta potential of 26.2∓4.1 mV and entrapment efficiency of POD of (88.56∓3.1)%. POD-NLC caused a significant inhibition of H8 cell proliferation in a concentration- and time-dependent manner. At an equivalent concentration, POD-NLC produced a stronger inhibitory effect on cell proliferation than POD. The inhibition rate of H8 cells after a 48-h exposure to POD-NLC and POD reached 95.8% and 65.6%, respectively, and at the highest concentration of 1 µg/ml, the IC(50) of POD-NLC and POD was 0.015 µg/ml and 0.13 µg/ml, respectively. Blank NLC did not obviously affect the proliferation of H8 cells. POD-NLC and POD both caused obvious increases in G(2)/M phase cell percentages and induced typical apoptotic changes of the cells, and their effects were comparable (P>0.05).</p><p><b>CONCLUSION</b>Compared with POD, POD-NLC has more potent effect in inhibiting H8 cell proliferation and inducing cell apoptosis, suggesting its potential in the treatment of cervical HPV infection.</p>

Mechanism of the reduction of cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB by propyl gallate / 药学学报

The probable mechanism of the reduction of rat cerebral ischemic-reperfusion injury by propyl gallate was studied. Intraluminal suture middle cerebral artery occlusion model of rat was employed. Propyl gallate was injected immediately after the ischemia was happened. The activity of NF-kappaB, and the expression of COX-2 and HSP70 on the peripheral ischemia were determined by Western blotting. The expression of TNF-alpha was determined by ELISA assay. RT-PCR and immunofluorescence staining were employed to detect the transcription and expression of TLR-4. Results showed that propyl gallate could inhibit the activity of NF-kappaB in the peripheral ischemia, and reduce the expression of COX-2 and TNF-alpha. As the upstream of NF-kappaB, the transcription and expression of TLR-4 decreased, as well as HSP70, the endogenic ligand of TLR-4. As an antioxidant, propyl gallate could reduce the cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB and decreasing the COX-2 and TNF-alpha in the peripheral ischemia. It also could influence HSP70 and TLR-4.

Quantitative determination of podophyllotoxin in dermal and blood microdialysis samples of rats by liquid chromatography-tandem mass spectrometry / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop and validate a sensitive method for quantitative analysis of podophyllotoxin in blood and dermal microdialysis samples of rats based on liquid chromatography-tandem mass spectrometry (UFLC-MS-MS).</p><p><b>METHODS</b>The microdialysis samples were prepared by liquid-liquid extraction using ethyl acetate with etoposide as the internal standard (IS). Podophyllotoxin was separated with an Agilent ZORBAX XDB-C18 column (2.1 mmx50 mm, 3.5 microm). The mobile phase consisted of acetonitrile: 10 mmol/L ammonium acetate (40:60, V/V) at a flow rate of 0.3 ml/min and the analysis was performed at the ambient temperature. The UFLC-MS/MS system was operated in the mode of multiple reaction monitoring using the electrospray ionization technique in positive mode.</p><p><b>RESULTS</b>Podophyllotoxin and etoposide responses were optimized at the transitions m/z 432.7-->397.3 and 589.5-->229.5, respectively. Calibration curves were linear over the range 2.0-1000 ng/ml. The lowest limits of quantification and detection values were 2.0 ng/ml and 0.7 ng/ml, respectively. The inter- and intra-day precision and accuracy were both less than 15%.</p><p><b>CONCLUSION</b>This selective and sensitive method can be used to quantity podophyllotoxin in the blood and dermal microdialysates of rats.</p>

Effect of wet dressing with Jingjielianqiao decoction on healing of acute radiation-induced skin ulcers in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the pharmacodynamics of Jingjielianqiao decoction in promoting the healing of acute radiation-induced skin ulcers in mice.</p><p><b>METHODS</b>Skin ulcers were induced in mice by radiation, and Jingjielianqiao decoction, rivanol or normal saline was applied for the treatment. The swelling of the legs was measured in mice using volumetric method, and the ulcer area after treatment was compared among the 3 groups.</p><p><b>RESULTS</b>No significant difference was found in the anti-inflammatory effects between Jingjielianqiao decoction group and rivanol group (P>0.05), but a significant difference was noted between Jingjielianqiao decoction group and normal saline group and also between rivanol group and normal saline group (P<0.01). Significant differences were observed in the ulcer healing among the 3 groups (P<0.01).</p><p><b>CONCLUSION</b>Jingjielianqiao decoction can significantly suppress radiation-induced acute inflammation and shows better effect than rivanol in promoting the healing of acute radiation-induced skin ulcer in rats.</p>

Preparation of lidocaine nanoemulsion and its transdermal absorption by rat skin ex vivo / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare lidocaine nanoemulsion and investigate its transdermal delivery ability in vitro.</p><p><b>METHODS</b>The optimal Km (surfactant/cosurfactant) value and the component proportion were determined by pseudoternary phase diagrams combined with Origin software analysis. The diameter and distribution range were detected by Zeta particle size analysis instrument, and the morphology of the nanoemulsion was observed by electron microscope. The permeation flux of lidocaine was determined in vitro using the modified Franz diffusion cell combined with HPLC, and the cumulative transdermal absorption amount and the apparent skin transdermal velocity were compared among nanoemulsion, gel and tincture containing 5% lidocaine. The permeation mode of lidocaine nanoemulsion was analyzed.</p><p><b>RESULTS</b>The average drop size of lidocaine nanoemulsion was 29.8-/+14.4 nm, and 98% of the drop sizes ranged from 15.1 to 45.5 nm and 2% from 77.9 to 261.3 nm. The nanoemulsion drop showed a spherical morphology in a polydisperse system. The Kp value of the nanoemulsion (3.07-/+0.74 cm/h) was significantly higher than that of gel (1.27-/+0.35 cm/h) and tincture (0.97-/+0.18 cm/h), and the permeation rate of the nanoemulsion was 69.82-/+7.48 microg x cm(-2) x h(-1), which fitted the the Zero-order release dynamic procedure.</p><p><b>CONCLUSIONS</b>The component proportion of lidocaine nanoemulsion can be conveniently obtained through pseudoternary phase diagrams and Origin software analysis, and the drop size, distribution, morphology and system type can be determined by Malvern Zetasizer combined with electron microscopy. The results also indicate that the nanoemulsion system with high permeation rate may provide a new promising means for local anesthesia.</p>

Effects of methotrexate on the expression of Toll like receptor(TLR)2 and TLR4 in human peripheral blood CD14~+ mononuclear cells from patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To investigate the expressions of TLR2 and TLR4 in patients with psoriasis vulgaris and the efrect of methotrexate(MTX)on them.so as to explore the therapeutic mechanism of MTX in psoriasis vulgafis.Methods Forty-three patients with psoriasis vulgaris were recruited into the study together with 30 normal human controls.Oral MTX was given to patients with an interval of 12 hours for three times per week until the control of conditions followed by 4 weeks of mainmining treatment.The dosage of MTX was 5 mg initially and decreased to 2.5 mg in the maintaining period.Flow cytometry was used to detect the expression of TLR2 and TLR4 in peripheral blood CD14~+ cells from the controls and patients at baseline,4 and 8 weeks after the beginning of treatment.Results The expression rate of TLR2 and TLR4 in CD14~+ cells was(92.6±4.3)%and(48.5±4.6)%,respectively,in untreated patients,significantly higher than that in normal controls(botll P＜0.01).A significant increase was observed in the expression rate of TLR2 and TLR4 in patients with active psoriasis compared with those with inactive psoriasis [(97.5±4.1)%vs(87.6±5.6)%,(55.3±5.8)%vs(40.7±7.1)%,both P＜0.05].Eigh weeks after the beginning of treatment with MTX.the expression rate of TLR2 and TLR4 significantly decreased to (79.6±6.7)%and(34.6±5.9)%.respectively(both P＜0.05).The psoriasis area and severity index(PASI)score had no significant correlation with the expression rate of TLR2 or TLR4(r=0.24.0.27,both P＞0.05).Conclusions TLR2,TLR4 and innate immune response mediated by both receptors play an important role in the pathogenesis of psoriasis.MTX may exert its therapeutic effect on psoriasis by inhibiting the expression of TLR2 and TLR4.