Heterocyclic compounds and phenolic glycosides from flowers of Dendrobium officinale / 中国中药杂志

Eight heterocyclic compounds and twelve phenolic glycosides were separated from the water extract of Dendrobium officinale flowers through chromatographic techniques, such as Diaion HP-20 macroporous adsorption resin column chromatography(CC), silica gel CC, ODS CC, Sephadex LH-20 CC, and preparative high performance liquid chromatography(PHPLC). According to the spectroscopic analyses(MS, ~1H-NMR, and ~(13)C-NMR) and optical rotation data, the compounds were identified as dendrofurfural A(1), 2'-deoxyadenosine(2), 4-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl] butanoic acid(3), 4-[2-formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoic acid(4), 1-(2-hydroxyethyl)-5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde(5), 5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde(6), methyl 5-(hydroxymethyl)-furan-2-carboxylate(7),(S)-5-hydroxymethyl-5H-furan-2-one(8), 2-methoxyphenyl-1-O-β-D-glucopyranoside(9), arbutin(10), isotachioside(11), 2,6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside(12), orcinol glucoside(13), tachioside(14), gastrodin(15), 4-O-β-D-glucopyranosylvanillyl alcohol(16), 2,6-dimethoxy-4-hydroxymethylphenol-1-O-β-D-glucopyranoside(17), icariside D_2(18), 4-formylphenyl-β-D-glucopyranoside(19), and vanillin-4-O-β-D-glucopyranoside(20). Among them, compound 1 is a new furfural benzyl alcohol condensate, with the skeleton first found in Dendrobium. Compounds 2-9, 11, 13, and 19 are reported from Dendrobium for the first time, and compounds 14 and 18 are reported for the first time from D. officinale. Compounds 11 and 14 showed moderate DPPH radical scavenging capacity, and compounds 11-14 demonstrated potent ABTS radical scavenging capacity, possessing antioxidant activity.

Erratum: Author correction to "The FAPα-activated prodrug Z-GP-DAVLBH inhibits the growth and pulmonary metastasis of osteosarcoma cells by suppressing the AXL pathway" Acta Pharm Sin B 12 (2022) 1288-1304

[This corrects the article DOI: 10.1016/j.apsb.2021.08.015.].

A new bibenzyl derivative from stems of Dendrobium officinale / 中国中药杂志

Eleven compounds were isolated from the 95% ethanol extract of the stems of Dendrobium officinale after water extraction by various modern chromatographic techniques, such as silica gel column chromatography(CC), octadecyl-silica(ODS) CC, Sephadex LH-20 CC, preparative thin layer chromatography(PTLC) and preparative high performance liquid chromatography(PHPLC). According to spectroscopic analyses(MS, 1D-NMR, 2D-NMR) combined with optical rotation data and calculated electronic circular dichroism(ECD), their structures were identified as dendrocandin Y(1), 4,4'-dihydroxybibenzyl(2), 3-hydroxy-4',5-dimethoxybibenzyl(3), 3,3'-dihydroxy-5-methoxybibenzyl(4), 3-hydroxy-3',4',5-trimethoxybibenzyl(5), crepidatin(6), alternariol(7), 4-hydroxy-3-methoxypropiophenone(8), 3-hydroxy-4,5-dimethoxypropiophenone(9), auriculatum A(10) and hyperalcohol(11). Among them, compound 1 was a new bibenzyl derivative; compounds 2 and 7-11 have not been previously reported from Dendrobium plants; compound 6 was reported from D.officinale for the first time. Compounds 3-6 exhibited potent antioxidant activity with IC_(50) values of 3.11-9.05 μmol·L~(-1) in ABTS radical scavenging assay. Compound 4 showed significant inhibitory effect on α-glucosidase, with IC_(50) value of 17.42 μmol·L~(-1), indicating that it boasted hypoglycemic activity.

The FAP α -activated prodrug Z-GP-DAVLBH inhibits the growth and pulmonary metastasis of osteosarcoma cells by suppressing the AXL pathway

Osteosarcoma is a kind of bone tumor with highly proliferative and invasive properties, a high incidence of pulmonary metastasis and a poor prognosis. Chemotherapy is the mainstay of treatment for osteosarcoma. Currently, there are no molecular targeted drugs approved for osteosarcoma treatment, particularly effective drugs for osteosarcoma with pulmonary metastases. It has been reported that fibroblast activation protein alpha (FAPα) is upregulated in osteosarcoma and critically associated with osteosarcoma progression and metastasis, demonstrating that FAPα-targeted agents might be a promising therapeutic strategy for osteosarcoma. In the present study, we reported that the FAPα-activated vinblastine prodrug Z-GP-DAVLBH exhibited potent antitumor activities against FAPα-positive osteosarcoma cells in vitro and in vivo. Z-GP-DAVLBH inhibited the growth and induced the apoptosis of osteosarcoma cells. Importantly, it also decreased the migration and invasion capacities and reversed epithelial-mesenchymal transition (EMT) of osteosarcoma cells in vitro and suppressed pulmonary metastasis of osteosarcoma xenografts in vivo. Mechanistically, Z-GP-DAVLBH suppressed the AXL/AKT/GSK-3β/β-catenin pathway, leading to inhibition of the growth and metastatic spread of osteosarcoma cells. These findings demonstrate that Z-GP-DAVLBH is a promising agent for the treatment of FAPα-positive osteosarcoma, particularly osteosarcoma with pulmonary metastases.

The physiological and psychological mechanisms of infra-slow oscillation / 生理学报

Infra-slow oscillation (ISO) is a kind of brain rhythm between 0.01 and 0.5 Hz. ISO is widely distributed in multiple brain regions. As an important psychophysiological activity, the ISO interacts with high-frequency neural rhythm via cross-frequency coupling, but has different activity patterns from high-frequency neural activity. Physiologically, the ISO may be generated by the dynamic activity of thalamus, glia, and ions. Psychologically, the frequency, amplitude, and phase of ISO could all regulate cognitive activities, but in different ways. Investigations on the ISO expands the neural rhythm research to lower frequency range, further promoting the construction of rhythmic theory of brain function.

Clinical effects of isotretinoin combined with anti-sensitive moisturizing tolerance-extreme cream on severe acne / 中华医学美学美容杂志

Objective:To observe the efficacy of anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin in the treatment of severe acne.Methods:Fifty patients with severe acne were selected in the Dermatology Clinic of the Third People＇s Hospital of Hangzhou from November 2018 to July 2019. They were randomly divided into the experimental group of 25 cases and the control group of 25 cases. The experimental group was treated with anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin orally. The control group was treated with isotretinoin orally alone. Before and after treatment for 56 days, lactate score, skin cuticle hydration (SCH), transepidermal water loss (TEWL) and skin physiological indexes were measured.Results:After 56 days of treatment, the TEWL and SCH of the control group were 15.75±3.31 and 10.13±3.62, the TEWL and SCH of the experimental group were 12.17±3.61 and 28.07±3.17, respectively; the difference was statistically significant ( T was 3.610 and 12.398, P was 0.002 and 0.000, respectively). The volume and depth of cyst nodule, scar depression, skin roughness, absolute value and area of erythema in the experimental group were significantly lower than those in the control group ( T was 2.280, 1.676, 2.332, 1.508, 4.813 and 3.637; P was 0.031, 0.011, 0.027, 0.040, 0.000 and 0.001, respectively). Conclusions:Anti-sensitive moisturizing tolerance-extreme cream combined with isotretinoin has a good effect on severe acne and it can reduce the barrier damage and other adverse reactions.

Effects of Loquat Lung-Cleaning Decoction on acne vulgaris with fusidic acid cream / 中华医学美学美容杂志

Objective:To investigate the clinical effect of Loquat Lung-Cleaning Decoction combined with fusidic acid cream in the treatment of acne vulgaris.Methods:From March 2019 to November 2019, 50 patients (9 males and 21 females, aged 13-31 years) with acne vulgaris were collected from the acne clinic of the dermatology department of the Hospital and divided into the experimental group and the control group with 25 cases in each group. The control group was treated with fusidic acid cream alone, and the experimental group was treated with Loquat Lung-Cleaning Decoction combined with fusidic acid cream. Six weeks after treatment, the clinical effect and adverse reactions were observed.Results:In the experimental group, 7 cases were cured, 10 cases were markedly effective, 6 cases were effective, but 2 cases were ineffective; the apparent efficiency was 68%. In the control group, 4 cases were cured, 6 cases were markedly effective, 9 cases were effective, and 6 cases were ineffective; the apparent efficiency was 40%. The effective rate of treatment in the experimental group was significantly higher than that in the control group, and the area of skin lesions was significantly smaller than that in the control group. The difference was statistically significant (rank sum test Z=1.98442, P=0.0472<0.05, rate analysis χ2 test χ2=3.945, P<0.05). Conclusions:The treatment of acne vulgaris with Loquat Lung-Cleaning Decoction and fusidic acid cream is effective.

Effects of dyclonine gel on success rate and complication of laryngeal mask airway insertion / 实用医学杂志

Objective To evaluate the effects of dyclonine gel on the success rate and complication of laryngeal mask airway(LMA)insertion. Methods Eighty ASAⅠ~Ⅱpatients undergoing ureteroscopic lithotripsy under general anesthesia received insertion of LMA coated with either dyclonine gel(group A,n=40)or paraffin oil(group B ,n = 40). Systolic blood pressure(SBP),diastolic blood pressure(DBP)and heart rate(HR)were recorded before anesthesia (T0),after induction (T1),at LMA insertion (T2),and 3 min after LMA insertion (T3). Time and frequency of LMA insertion ,incidence of abnormal sensation of pharynx and sore throat after surgery in 2 groups were recorded. Results Compared with those at T0,the SBP,DBP and HR were lower at T1, T2 and T3 in 2 groups(P<0.05). The one-time success rate of group A and group B were 90%and 97.5%respec-tively(P>0.05).The incidence of abnormal sensation of pharynx and sore throat was significantly lower in group A (5%vs. 20%)(P<0.05). Conclusion Dyclonine gel cream-coated LMA can reduce the incidence of sore throat in patients undergoing ureteroscopic lithotripsy under general anesthesia.

Clinical study of local application of triamcinolone acetonide in promoting wound healing after granulomatous lobular mastitis operation / 中国医师进修杂志

Objective To investigate the effect of local application of triamcinolone acetonide in promoting wound healing and recurrence after granulomatous lobular mastitis (GLM) operation. Methods The clinical data of 55 GLM patients with the diameter > 5 cm were retrospectively analyzed, and all patients treated with incision drainage to breast abscess and removal of inflammatory lesions. Thirty - one patients were treated with routine dressing change after operation (control group), and 24 patients were treated with routine dressing change and triamcinolone acetonide injection after operation (experiment group). The wound dressing time, recovery rate and recurrence rate were compared between 2 groups, and hormone related adverse reaction of experiment group was observed. Results The wound dressing time in experiment group was significantly shorter than that in control group:(58 ± 19) d vs. (71 ±25) d, and there was statistical difference (P0.05. The patients of experiment group did not have the hormone related adverse reaction. Conclusions The local application of triamcinolone acetonide after GLM operation can accelerate the wound healing, and it may also contribute to higher recovery rate and lower recurrence rate.

Effects of dyclonine gel on success rate and complication of laryngeal mask airway insertion / 实用医学杂志

Objective To evaluate the effects of dyclonine gel on the success rate and complication of laryngeal mask airway(LMA)insertion. Methods Eighty ASAⅠ~Ⅱpatients undergoing ureteroscopic lithotripsy under general anesthesia received insertion of LMA coated with either dyclonine gel(group A,n=40)or paraffin oil(group B ,n = 40). Systolic blood pressure(SBP),diastolic blood pressure(DBP)and heart rate(HR)were recorded before anesthesia (T0),after induction (T1),at LMA insertion (T2),and 3 min after LMA insertion (T3). Time and frequency of LMA insertion ,incidence of abnormal sensation of pharynx and sore throat after surgery in 2 groups were recorded. Results Compared with those at T0,the SBP,DBP and HR were lower at T1, T2 and T3 in 2 groups(P<0.05). The one-time success rate of group A and group B were 90%and 97.5%respec-tively(P>0.05).The incidence of abnormal sensation of pharynx and sore throat was significantly lower in group A (5%vs. 20%)(P<0.05). Conclusion Dyclonine gel cream-coated LMA can reduce the incidence of sore throat in patients undergoing ureteroscopic lithotripsy under general anesthesia.

Effects of Yishen Tonglong Capsules on sex hormone levels in mice with benign prostatic hyperplasia / 中华男科学杂志

<p><b>Objective</b>To study the effect of Yishen Tonglong Capsules (YTC) on the sex hormone levels in the mouse model of benign prostatic hyperplasia (BPH).</p><p><b>METHODS</b>The BPH model was made in male mice by subcutaneous injection of testosterone propionate at 5 mg per kg of the body weight per day for 3 weeks. Then the model animals were divided into five groups of 10 in each: model control, Longbishu Capsules (LBS), and high-, medium- and low-dose YTC. Another 10 mice were included as normal controls. The mice in the LBS group were treated intragastrically with LBS at 0.45 g per kg of the body weight, those in the high-, medium- and low-dose YTC groups with YTC at 1.2, 0.6 and 0.3 g per kg of the body weight, and those in the model and normal control groups given the same volume of distilled water. After 8 weeks of treatment, the mice were sacrificed and their prostates taken for measurement of their wet weight, calculation of the prostatic index (PI), determination of the levels of serum testosterone (T), estradiol (E2) and E2/T ratio, and observation of the morphological changes of the prostate tissue under the light microscope.</p><p><b>RESULTS</b>The wet weight of the prostate and PI were significantly decreased in the LBS and medium- and high-dose YTC groups as compared with the model control group (P<0.01). The serum T and E2 levels and E2/T ratio were (1.73±0.02) ng/ml, (73.08±1.03) pg/ml and 42.30±0.53 in the normal control, (3.86±0.02) ng/ml, (145.79±0.88) pg/ml and 37.76±0.25 in the model control, (2.47±0.02) ng/ml, (95.87±0.47) pg/ml and 38.80±0.13 in the LBS, (2.91±0.03) ng/ml, (112.68±0.77) pg/ml and 38.80±0.42 in the low-dose YTC, (2.77±0.02) ng/ml, (112.16±0.82) pg/ml and 40.56±0.29 in the medium-dose YTC, and (2.75±0.03) ng/ml, (107.11±0.61) pg/ml and 38.92±0.36 in the high-dose YTC group, all with statistically significant differences between the model control and the other groups (P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>Yishen Tonglong Capsules significantly reduced the levels of serum T and E2 and elevated the E2/T ratio in the mouse model of BPH, which manifested the action mechanism of Yishen Tonglong Capsules in the treatment of BPH.</p>

Etiologies and clinical features of 19 cases with bilateral acute sensorineural hearing loss / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the etiologies and clinical features for bilateral acute sensorineural hearing loss (bi-ASNHL).</p><p><b>METHODS</b>The clinical data of 19 cases presenting with bi-ASNHL were retrospectively analyzed, including the clinical features, systemic examinations, laboratory examinations, audiology and radiology results, as well as the prognosis.</p><p><b>RESULTS</b>There were 15 non-otologic diseases in 19 patients, accounting for 78.9% of the total cases, most of which were disorders with multisystem and multi-organ disorder. The central nervous system diseases including fungal meningitis, tuberculous meningitis, and viral encephalitis in 3 patients. The clinical features of deafness were bilateral, progressive, accompanied with fever, headache, dizziness, nausea, vomiting and change of mental status. There was a decrease in speech recognition score (SRS), and speech recognition threshold (SRT) was obviously inferior to pure tone average (PTA) disproportionally. Diseases of immune system including antineural cytoplasmic antibody (ANCA)-associated systemic vasculitis (AASV), relapsing polychondritis (RP), rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE) in 5 patients. They showed the characteristics of bilateral, progressive and simultaneous autoimmune disease. Hematological and endocrine system diseases including diabetes mellitus, leukemia, and thyroid hypofunction in 5 patients. The deafness had the characteristics of symmetry and progressivity. Otologic diseases including large vestibular aqueduct syndrome (LVAS) and sudden sensorineural hearing loss (SSNHL) in 4 patients; Drug-induced sensorineural deafness happened in 2 patients. After the treatment aimed at the causes, 1 case was cured, 3 patients were markedly effective, 7 patients were effective, and 8 patients were ineffective(including dead and refusal cases), with a total effective rate of 57.9%.</p><p><b>CONCLUSIONS</b>The most of bi-ASNHL cases are often associated with systemic diseases. Clinicians should analyze the history and clinical characteristics in detail, and complete specific laboratory examinations, audiology and imaging examinations in order to reveal the causative diseases. It should be treated aimed at the etiology.</p>

The Effects of Phonosurgery Combined with Voice Therapy for Vocal Polyps Accompanied with Muscle Tension Dysphonia / 听力学及言语疾病杂志

Objective To evaluate the effects of phonosurgery combined with voice therapy in patients with vocal polyps accompanied with muscle tension dysphonia (MTD) .Methods The study retrospectively enrolled 117 patients with a diagnosis of vocal polyps accompanied with MTD .All the patients were divided into two groups ac‐cording to the different remedies:the surgery group (n=57) ,and the combination group (n=60) .The Xion acous‐tic analysis was carried out on the two groups before and after therapy .We compared the results in each group ,the results of post-therapy between two groups ,and that of post-therapy with the control group(n=50) .The acous‐tic parameters were jitter percent (jitter) ,shimmer percent (shimmer) ,dysphonia severity index (DSI) .Results The results obtained in the study showed that all the voice analysis parameters were significantly improved after therapy in both the surgery group and combination group(P0 .05) .Conclusion As for vocal polyps accompanied with M TD ,successful phonosurgical treatment and voice therapy can improve patients’ voice quality .

Application of two different gastrectomy methods in proximal gastric cancer / 国际肿瘤学杂志

Objective To evaluate the impacts of the two different gastrectomy methods on the quality of life,complication and prognosis in proximal gastric cancer.Methods One hundred and two cases of proxi-mal gastric cancer in Tung Wah Hospital were collected for retrospective analysis.They were divided into proxi-mal gastrectomy/gastroesophagostomy (PG)group (n =50)and total gastrectomy/esophagojejunostomy (TG) group (n =52),according to the methods of gastrectomy and reconstruction.The postoperative complications, nutritional status and prognosis of the two groups were compared.Results The incidence of reflux esophagitis was obviously higher in PG group than that in TG group (38.0% vs 1 9.2%,χ2 =4.464,P =0.035).No sig-nificant differences were found between the two groups in the incidences of postoperative infection,bleeding and anastomotic leakage (χ2 =0.063,P =1 .000;χ2 =0.001 ,P =0.978;χ2 =0.31 1 ,P =0.577).There were no significant differences between PG and TG group in total plasma protein [(65.26 ±4.1 0)g/L vs (65.33 ± 3.75)g/L,t =-0.402,P =0.688],albumin [(39.76 ±2.1 7)g/L vs (39.59 ±2.04)g/L,t =1 .778,P =0.076],hemoglobin [(1 07.33 ±1 1 .1 0)g/L vs (1 08.09 ±1 1 .1 7)g/L,t =-1 .502,P =0.1 33]and weight loss [1 .00 ~8.00 kg vs 0.50 ~8.20 kg,t =-1 .622,P =0.1 05]in one year postoperatively.All cases were followed-up for 7 months to 1 0 years.No significant differences were found between PG and TG group in the incidences of anastomotic tumor recurrence (4.0% vs 5.8%,χ2 =0.1 71 ,P =0.679),metastasis (24.0% vs 28.8%,χ2 =0.308,P =0.579)and median survival time (53.6 months vs 49.8 months,χ2 =2.564,P =0.1 09).Conclusion Compared with PG group,the incidence of postoperative reflux esophagitis is effectively reduced,and the incidences of malnutrition,tumor recurrence and metastasis and death are not increased in TG group.Hence,TG should be a safe and effective surgery strategy.

Experimental study on the impact of photodynamic therapy on the tissue structure of normal nasal mucosa in rabbit / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the reactive changes of normal nasal mucosa in rabbit under photodynamic therapy (PDT) and to make a preliminary research for the treatment of allergic rhinitis (AR) with PDT.</p><p><b>METHODS</b>Twenty-four New Zealand rabbits were divided into 2 groups, an experimental group and a control group with 12 rabbits in each group. PDT was applied to the experimental group, while the control group was given no treatment. The nasal mucosa was sampled separately from the same position of the rabbits from the 2 groups on 1st, 3rd, 7th, 14th, 21st, 28th day. Histomorphological changes of the sampled nasal mucosa were observed under light microscope and transmission electron microscope (TEM). The damage of three tachykinins: substance P (SP), neurokinin A (NKA) and neurokinin B (NKB) of nerve fibers was observed after immunohistochemical staining.</p><p><b>RESULTS</b>Compared with the control group, the nasal mucosa tissues from the experimental group had serious inflammatory reaction with basal layer damaged on the 1st and 3rd day after PDT application, the epithelial cells of nasal mucosa were arranged in disorder, and part of cilium shortened and became abnormal or even disappeared, each organelles damaged obviously; on the 7th, 14th, 21st day, it could be seen that ciliated cell, columnar cell and goblet cell started regeneration, basal cell and lamina propria glands proliferated, and the glands appeared secretion phenomenon; on the 28th day, ciliated columnar epithelium took back the nasal mucosas with small amount of microvilli, and mucous granules were found in the column cells. Nasal immunohistochemical staining of the experimental group from various stages showed that three kinds of neuropeptides were not expressed.</p><p><b>CONCLUSIONS</b>Normal rabbit nasal mucosas will be temporarily damaged after PDT application, the damaged nasal mucosa begin to recover in one week, and return to normal in about four weeks. Most structure and functions have recovered at the fourth week except some nerve endings.</p>

Experimental study on the treatment of allergic rhinitis with photodynamic therapy / 国际生物医学工程杂志

Objective To preliminarily investigate the therapeutic effect of photodynamic therapy (PT) treating allergic rhinitis (AR).Methods Eighteen New Zealand rabbits were randomly divided into 3 groups,experimental group A,negative control group B and blank control group C.Rabbits of A and B groups were sensitized with ovalbumin (OVA) to get animal AR model.Group A was treated with PDT while group B and C were applied with natural light treatment.Symptoms and signs before and after treatment were compared and the change of nasal mucosa was observed with light microscope and transmission electron microscope.Results After PDT treatment,group A rabbits' AR symptoms and signs were improved significantly compared to group B,light microscope and transmission electron microscope observation showed that the number of inflammatory cells of group A rabbits decreased significantly and the mucosal structure returned to normal after PDT treatment.After 21 d,as compared with group C,the differences of the times of sneezing and scratching and nasal secretion had no statistical significance (P＞0.05).Conclusions PDT has certain therapeutic effect in experimental AR treatment,which can be used in AR treatment after consummating its therapeutic mechanism.

Resonance Characteristics of Pre-and Post -UPPP Operation in Patients with OSAHS / 听力学及言语疾病杂志

Objective To study the influence of UPPP on resonance characteristics of four resonance cavity in patients with OSAHS .Methods Using multi-channel voice analysis system to test the cavity resonance character-istics and energy distribution of 36 OSAHS patients (including pre - and post -operative) and 36 normal males when they speak /a:/.The frequency spectrum was 4 000Hz(FR1 ,FR2 ,FR3 ,FR4 ) .Results FR2 energy values of the head cavity and mouth cavity of OSAHS patients heighten significantly after UPPP (P<0 .05);FR1 energy val-ues of head ,mouth and thoracic cavity were higher than that of in normal males'after UPPP(P<0 .05) .FR2 ,FR3 , FR4 energy values of head cavity ,FR2 ,FR3 energy values of mouth cavity ,FR2 energy values of thoracic cavity were lower than those of in normal males'after UPPP(P<0 .05) .Conclusion UPPP surgery removes the obstruction of the upper airway of OSAHS patients and changes the morphology of the pharynx ,thas changes the resonance charac-teristics of the head cavity and oral .

Effects of calcipotriol on melanocytes and CD8 + cytotoxic T lymphocytes from patients with vitiligo / 中华皮肤科杂志

Objective To evaluate the effect of calcipotriol on the proliferation of and cytokine secretion by melanocytes and perilesional CD8+ cytotoxic T lymphocytes (CTLs) from patients with vitiligo.Methods Melanocytes isolated from abdominal skin and CD8+ CTLs from perilesional skin of patients with vitiligo were subjected to successive culture in vitro.After several passages,the melanocytes and CD8+ CTLs were cultured alone or in combination with or without the presence of various concentrations of calcipotriol for 24 to 48 hours.MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium,inner salt) method was used to evaluate the proliferative activity of cells,enzyme-linked immunosorbent assay to determine the levels of interleukin (IL)-6,tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supematant of cells,flow cytometry to detect cell apoptosis.Some co-cultured melanocytes and CTLs were treated with calcipotriol of 10-8 mol/L and anti-IL-6 antibody of various concentrations (0,1,2,2.5,5,10 mg/L) for two days followed by enumeration of cells.The concentrations of 108 and 10-9 mol/L (calcipotriol) were chosen for relevant tests.Results There was a marked apoptosis in MCs after coculture with CD8+ CTLs.The 24-hour treatment with calcipotriol of 104 and 10-9 mol/L had no obvious effect on the proliferation of melanocytes cultured alone (both P ＞ 0.05),but accelerated the proliferation of melanocytes cocultured with CTLs (both P ＜0.05) as well as that of CD8+ CTLs cultured alone or in combination with melanocytes (all P ＜0.05).A statistical decrease was observed in IL-6,TNF-α and IFN-γlevels in the supernatant of cocultured melanocytes and CTLs compared with those in the supernatant of melanocytes and CTLs cultured alone,and calcipotriol of 10-9 mol/L intensified the decrease in supernatant IL-6 level (t =2.89,P ＜0.05),but no statistical changes were noted for the level of TNF-α or IFN-γin the supernatant of the coculture system after treatment with calcipotriol of 104 or 104 mol/L compared with that before treatment (both P ＞ 0.05).In the coculture system pretreated with calcipotriol of 10-8 mol/L,the number of CD8+ CTLs significantly decreased (t =3.15,P ＜0.05),whereas that of melanocytes significantly increased (t =3.53,P ＜0.05) after the treatment with anti-IL-6 antibody of 5 mg/L.Conclusions Perilesional CD8+ CTLs have a specific cytotoxic effect on melanocytes,and calcipotriol may inhibit the cytotoxic effect of CD8+ CTLs by suppressing the secretion of IL-6,which may partly explain the therapeutic mechanism of calcipotriol for vitiligo.

In vitro effects of tea polyphenols on the secretion of TNF-α, IFN-γ and IL-2R by peripheral blood CD8+ T lymphocytes from vitiligo patients / 中华皮肤科杂志

Objective To detect the level of cytokines and their receptors secreted by peripheral blood CD8 + T lymphocytes from vitiligo patients,and to evaluate the influence of tea polyphenols on their secretion.Methods CD8+ T lymphocytes were isolated from the peripheral blood of 12 patients with progressive vitiligo,12 patients with stable vitiligo and 10 healthy controls,and cultured in vitro for 20 days.Then,some CD8+ T lymphocytes were treated with tea polyphenols of 100 mg/L for two days.Enzyme-linked immunosorbent assay (ELISA) was carried out to determine the levels of tumor necrosis factor (TNF)-α,interferon (IFN)-γ and interleukin-2 receptor (IL-2R) in the culture supernatants of CD8+ T lymphocytes before and after the treatment with tea polyphenols.Analysis of variance and t test were conducted to assess differences in these parameters among these groups and changes between pretreatment and posttreatment,respectively.Results There was a sequential decrease in the level of TNF-α ((191.302 ± 6.077) vs.(175.966 ± 2.467) vs.(173.664 ± 3.600) ng/L,F =4.784,P ＜ 0.05),but a sequential increase in that of IFN-γ ((280.182 ± 36.070) vs.(371.670 ± 24.352) vs.(447.147 ± 8.432) ng/L,F=9.036,P＜ 0.01) and IL-2R ((8.375 ± 0.161) vs.(8.845 ± 0.161) vs.(9.345 ±0.125) ng/L,F =9.639,P ＜ 0.01) in the culture supernatant of CD8+ T lymphocytes from patients with progressive vitiligo to patients with stable vitiligo and healthy controls.After two days of tea polyphenol treatment,a statistical decrease was observed in the level of TNF-α in the culture supernatant of CD8 + T lymphocytes from patients with progressive vitiligo ((164.797 ± 1.784) ng/L,P ＜ 0.01),patients with stable vitiligo ((166.150 ± 3.576) ng/L,P ＜ 0.05) and healthy controls ((155.028 ± 5.759) ng/L,P ＜ 0.05),but no statistical changes were noted for IFN-γor IL-2R (all P ＞ 0.05) despite a slight elevation in the level of IFN-γin the culture supernatant of CD8+ T lymphocytes from patients with stable vitiligo and in that of IL-2R from all the three groups as well as a mild reduction in that of IFN-γ from patients with progressive vitiligo and healthy controls.Conclusions The changes of cytokines and their receptors secreted by CD8 + T lymphocytes may be associated with the induction and progression of vitiligo.Tea polyphenols may treat vitiligo via suppressing the secretion of TNF-α by CD8+ T lymphocytes.

Protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T ceils from vitiligo patients / 中华皮肤科杂志

Objective To evaluate the protective effects of tea polyphenols against the destruction of melanocytes by CD8+ T cells from vitiligo patients.Methods Skin tissue was resected from the margin of vitiligo lesions followed by the isolation and culture of CD8+ T lymphocytes,and from the normal skin of vitiligo patients followed by the isolation and culture of melanocytes.Flow cytometry was carried out to evaluate the purity of CD8+ T cells.The melanocytes were cocultured with the CD8 + T cells at different ratios followed by the evaluation of killing effect of CD8+ T cells.Various concentrations (200 and 400 μg/ml) of tea polyphenols were added into the co-culture system of CD8+ T cells and melanocytes at a ratio of 5 ∶ 1 followed by an additional culture of 48 hours.Then,flow cytometry was performed to detect the apoptosis in melanocytes in the coculture system.Results CD8+ T lymphocytes were successfully obtained from the marginal area of vitiligo lesions with a purity of more than 90％,which highly expressed the antigens CD137 and CD69.The coculture with CD8+ T cells markedly accelerated the apoptosis in melanocytes,while the accelerative effect was inhibited by tea polyphenols of 200 and 400 μg/ml.Conclusions The CD8+ T cells infiltrating the edge of vitiligo lesions display a potential destructive effect on autologous melanocytes from vitiligo patients,and tea polyphenols have a protective effect against the destruction of melanocytes by CD8+ T cells.