| Blackheads and coarse pores can seriously impact facial beauty and reduce the self-confidence of patients.There are 3 major clinical causes of enlarged facial pores,namely high sebum excretion,decreased elasticity around pores,and increased hair follicle volume.Given the different possible causes for enlarged pores,therapeutic modalities must be individualized for each patient.The traditional treatment of acne include needle-lancing and topical drugs,such as retinoic acid and salicylic acid,However,conventional cleaning products are ineffective for certain blackheads.Evidence suggests that Q-switched Nd:YAG and lattice laser treatment for blackheads and coarse pores is effective,but the improvement was limited and the effect was not achieve lasting stability.Blackheads and coarse pores on nose with nasal hair follicles and sebaceous gland cell aggregated excessively in the dermis,dysregulation of cell proliferation and apoptosis plays an important key role in the pathogenesis and progression of nasal blackheads and coarse pores in patients.That is,the imbalance of proliferation and apoptosis of sebaceous gland cells and PCNA may be an important factor in the development of the nose blackheads and the coarse pores.The process of apoptosis is tightly regulated to many genes,including oncogenes,tumor suppressor genes and virus genes.These genes such as Bcl-2 family,caspase family,C-myc,cancer genes such as tumor suppressor genes such as P53.Bcl-2 family is the earliest study of apoptosis related genes,Bcl-2,Bcl-xl,Mcl and A1 genes belong to anti apoptosis genes.Bax,Bak and bcl-xS genes,which belong to the pro apoptotic gene.It is known that apoptosis may have three main signal transduction pathways: control pathways on which the p53 gene depends,mitochondrial pathways,and death receptor pathways of apoptosis.Caspase-3is a factor in performing apoptosis,which can cause a cascade amplification effect of downstream enzymes once triggered.Its excitation can lead apoptosis to an apoptosis stage that is related to nuclear change and is the most important downstream effect of protease.In the signal transduction pathway of cell apoptosis,the component ratio of Bcl-2 family members is the key factor of apoptosis regulation,especially the Bcl-2/Bax ratio in the molecular switch that can trigger apoptosis.When Bcl-2/Bax ratio decreased molecular switch to initiate the apoptosis program.Bcl-2 and Bax regulate apoptosis by forming homodimers or heterodimer,when Bcl-2 and Bax forms homodimer to induce apoptosis,Bcl-2 and Bax may form a heterodimer inhibit apoptosis.In recent years,more and more studies have been done on the effects of light or laser induced cell apoptosis.Pulsed dye laser,carbon dioxide laser and He-Ne laser have been reported to induce apoptosis.800nm semiconductor laser has been used for a long time on clinic,for higher safety and minor side effects,it has been widely applied in skin cosmetology areas and proved to be effective.To observe the effect of the function that applied with 800 nm semiconductor laser on the back skin of guinea pig hair follicle and sebaceous gland.All of above is for the clinical treatment of skin blackhead pores and provide a theoretical basis for the clinical treatment of skin blackhead pores.There is no research on using of800 nm semiconductor laser treatment of skin blackheads and coarse pores nationally and internationally.Part one Morphological observation on the effect and the change ofPCNA that the function of the treatment and of 800nm semiconductor laser on the black skin guinea pig hair follicle and sebaceous gland.Objective: To study the changes of 800 nm semiconductor laser on the black skin of guinea pig hair follicle and sebaceous gland histopathology and the function on protein expression of sebaceous gland cell proliferation cell nuclear antigen(PCNA).Methods: A total of 24 healthy short-haired male guinea pigs were evenly coated with coal tar suspension,an experimental area of 8cm × 3cm on the back of the guinea pigs.Divided into four groups: normal control group(NC),low-dose 800 nm semiconductor laser treatment group(L-LS),high-dose 800 nm semiconductor laser treatment group(H-LS),and Q-switched Nd:YAG treatment group(QC).Samples were taken respectively at 1d,7d,14 days after surgery.Observation of the changes of epidermis,dermis,sebaceous glands and hair follicle damage by HE staining,and the expression of the proliferating cell nuclear antigen(PCNA)was detected by immunohistochemistry.Results: HE staining showed that 800 nm semiconductor laser and Qswitched Nd:YAG laser treatment,compared with the normal control group:both the hair follicle distortion,degeneration,hair papilla and hair ball separation occurred.Inflammatory cell infiltration around the epidermis and the hair follicle.Dermis collagen fiber can be seen swelling but not break.14 d can be seen hair follicle epithelium become thinning.The number and the volume of hair follicles and sebaceous glands are decreased,and the reduction of inflammatory cells,and the morphology returned to normal.The reduction of hair follicles and sebaceous glands in high-dose Light Sheer Duet 800 nm laser group was the most obvious.Immunohistochemical staining showed that black dorsal skin PCNA positive particles were mainly distributed in the nuclei of epidermal basal layer cells,hair follicle epithelial cells and stromal cells.At each time period,expression of PCNA positive particles of black guinea pig dorsal skin in NC group(1d 32.6±5.5% 、 7d 33.5±5.4% 、 14 d 32.7±5.3%)were the highest.Followed by L-LS group(25.5±6.3%、7d 23.2±6.2%、14d 21.7±6.0%)and QC group(1d 15.4±4.5% 、 7d 14.3±4.6% 、 14 d 12.8±4.8%),H-LS group(1d11.2±4.7%、7d 8.6±4.2%、14d 6.3±4.3%)were the lowest,with the difference being statistically significant(1d F=12.304、7d F=13.655、14d F=15.234,P<0.001).And in 1st day to 14 th day the expression levels of PCNA in each groups were reduced with time,with the difference being statisticallysignificant(P < 0.05).The above results show that Light Sheer Duet 800 nm semiconductor laser therapy can effectively reduce the number and volume of hair follicles and sebaceous glands of the black skin guinea pig,and does not lead to damage dermal elastic fiber fracture,Light Sheer Duet 800 nm high-dose semiconductor laser treatment group was better than that of the low-dose group and Q-switched laser treatment group.Part two Effect on the apoptosis related genes that treatment on 800nm semiconductor laser sebaceous gland cell of black skin of guinea pigObjective: To study the effect of 800 nm semiconductor laser on the skin of guinea pig hair cell apoptosis and expression of Caspase-3,Bax,Bcl-2protein.A total of 24 healthy short-haired male guinea pigs were evenly coated with coal tar suspension,an experimental area of 8cm×3cm on the back of the guinea pigs.Divided into four groups: normal control group(NC),low-dose 800 nm semiconductor laser treatment group(L-LS),high-dose800 nm semiconductor laser treatment group(H-LS),and Q-switched Nd:YAG treatment group(QC).Samples were taken respectively at 1d,7d,14 days after surgery.TUNEL method was used to detect the apoptosis of guinea pig hair back skin sebaceous gland cells,Western-blot was used to detect the expression of Caspase-3,Bax,Bcl-2 protein.Results: TUNEL method was used to detect the black skin of guinea pig sebaceous cell apoptosis,and brown staining were TUNEL(+)cells,TUNEL(+)cells were mainly distributed in the basal layer of the epidermis and hair follicle epithelial layer and stromal layer.Cell apoptosis rate of black dorsal skin in NC group(1d 0.5±0.1%,7d 0.4±0.1%,14 d 0.6±0.2%)was the lowest,followed by L-LS group(1d 6.7±1.2%,7d 7.6±1.3%,14 d 8.2±1.5%)and QC group(1d 8.2±2.0%,7d 9.3±2.2%,14 d 10.2±2.3%),TUNEL(+)cells in H-LS group(1d 13.6±3.4%,7d 15.8±3.5%,14 d 17.2±3.6%)was the highest,with the difference being statistically significant(1d F=10.235,7dF=11.527,14 d F=14.520,P < 0.001).And from 1st day to 14 th day the TUNEL(+)cells in each groups were increased with time,with the difference being statistically significant(P < 0.05).Western blot was used to detect the expression level of caspase-3 protein in black skin of guinea pigs.The expression of caspase-3 protein was compared with that of GAPDH,which was the relative expression level of protein.Expression level of caspase-3 protein of black dorsal skin in NC group(1d 0.21±0.06,7d 0.22±0.05,14 d 0.20±0.07)was the lowest.In each period,the relative expression level of caspase-3 protein was increased by laser treatment.The relative expression level of caspase-3 protein in H-LS group(1d 0.47 ± 0.13,7d 0.50 ± 0.16,14 d 0.55 ± 0.18)was the highest.Followed by QC group(1d 0.44±0.20,7d 0.48±0.21,14 d 0.52±0.24).The relative expression level of caspase-3 protein was higher.And then for the L-LS group(1d 0.42±0.10,7d 0.46±0.12,14 d 0.50±0.14),And each group had significant difference compared with NC group(1d F=8.632,7d F=9.203,14 d F=9.624,P<0.001).And from 1st day to 14 th day the expression level of Caspase-3 protein in L-LS group,H-LS group and QC group was increased with time,with the difference being statistically significant(P < 0.05).In each period,laser treatment could up-regulate the relative expression level of Bax protein and down-regulate of Bcl-2 protein.The relative expression level of Bax protein was the highest in the high energy treatment group(H-LS)with Light Sheer Duet 800 nm semiconductor laser,the relative expression level of Bcl-2 protein was the lowest,and the ratio of Bcl-2/Bax was the lowest.Followed by QC group,up-regulate the relative expression level of Bax protein and down-regulate of Bcl-2 protein was higher.And then for L-LS group.NC group(Bax 1d 0.15±0.04,7d 0.16±0.03,Bcl-2 14 d 0.14±0.05;1d 0.23±0.08,7d 0.24±0.06,14 d 0.23±0.07)was the lowest.1d: In L-LS group the relative expression of Bax protein in black skin of guinea pigs was 0.36±0.10,Bcl-2 was 0.16±0.10;In QC group Bax was 0.40±0.21,Bcl-2was 0.14±0.05;In H-LS group was the highest expression level of Bax protein was 0.42±0.14,Bcl-2 was 0.10±0.03.Compared with the NC groupdifferences were statistically significant(Bax F=9.032 P < 0.001;Bcl-2F=9.562 P < 0.001).7d: In L-LS group the relative expression of Bax protein in black skin of guinea pigs was 0.40±0.13,Bcl-2 was 0.18±0.06;In QC group Bax was 0.43±0.22,Bcl-2 was 0.16±0.06;In H-LS group was the highest expression level of Bax protein was 0.46±0.13,Bcl-2 was 0.12±0.04.Compared with the NC group differences were statistically significant(Bax F=9.421 P<0.001;Bcl-2 F=9.624 P<0.001).14d: In L-LS group the relative expression of Bax protein in black skin of guinea pigs was 0.43±0.11,Bcl-2was 0.20±0.09;In QC group Bax was 0.45±0.23,Bcl-2 was 0.18±0.05;In H-LS group was the highest expression level of Bax protein was 0.49±0.16,Bcl-2 was 0.14±0.04.Compared with the NC group differences were statistically significant(Bax F=9.637 P<0.001;Bcl-2 F=9.758 P<0.001).And from 1st day to 14 th day up-regulate the relative expression level of Bax protein and down-regulate of Bcl-2 protein in L-LS group,H-LS group and QC group was increased with time,with the difference being statistically significant(P < 0.05).The above results show that Light Sheer Duet 800 nm semiconductor laser can promote the apoptosis of black guinea-pig skin sebaceous gland cells,through up-regulation expression of Bax protein and down-regulation of Bcl-2protein,decreasing Bcl-2/Bax ratio,to up-regulate the activation of caspase-3.Part three Study on the effect of 800 nm semiconductor laser irradiation on the improvement of nose blackheads and coarse poresObjective: Evaluation of the efficacy,satisfaction and safety of 800 nm semiconductor laser in the treatment of nasal blackheads and coarse pores.Methods: This study recruited 60 volunteers,all volunteers on nose tip or nosewing position appears different degrees of coarse pores and blackheads phenomenon.A total of 30 patients in the observation group were treated with Light Sheer ET 800 nm semiconductor laser.A total of 30 patients in the control group were treated with Q-switched Nd:YAG laser.Evaluation of facial coarse pore score(enlarged facial pores score,EFPS).The application of CANON D8000 SLR camera to the observation group andthe control group before and after treatment of the nose tip and nose on both sides of the blackhead and pores.The application of CANON D8000 SLR camera to take macro photo shoot before and after treatment to the observation group and the control group of the nose tip and both sides of nosewing of the blackhead and coarse pores.And determine according to photos that the the difference to the pores size,density and depth.The level of the patient’s nose blackhead and coarse pores were classified according to 1-6grade.Determine the average difference of the nose tip and both sides of nosewing of the blackhead and coarse pores before and after treatment.Improvement rate =(pre treatment rating-post treatment rating)/ pre treatment rating ×100%.Improvement rate = significant improvement rate +effective improvement rate.Subjective assessment by patients that the improvement situation of their nose pores and blackhead after treatment.The criteria are divided into three levels of very satisfied,satisfied and dis satisfied.The satisfaction rate is the sum of very satisfaction rate and satisfaction rate.The incidence of adverse reactions,including epidermal callus removal duration,erythema duration and pain swelling duration.The visual analogue scale VAS(visual analogue scale)was used to assess the degree of pain.The most painful for 10 points,painless for the 0 points.Volunteers according to their own pain situation to choose the score after the end of the treatment.Results: To evaluate the enlarged facial pores score of the patients in the observation group were treated with Light Sheer ET 800 nm semiconductor laser and the control group treated with Q-swtiched Nd:YAG laser before and after treatment.The observation group of volunteers of the nose pores and blackheads improvement rate was 93.3%,control group was 76.7%,with the difference being statistically significant(P < 0.05).Assessment of the recovery,medical treatment technology,service attitude and other aspects after the end of the treatment in both observation group and control group,out of 100 points.There were 10 patients in the control group were very satisfied,accounting for more than 33.3%;11 cases were satisfied,accounting for 36.7%;9 cases were not satisfied with theproportion of 30.3%;Total satisfaction was 70%.In the observation group,17 cases were very satisfied,10 cases were satisfied,3 cases were not satisfied.The proportion was 56.7%,33.3% and 10%,respectively;Total satisfaction was 90%.It can be seen that the two groups of volunteers for the treatment of satisfaction with the results of a larger difference,with statistical significance(P < 0.05).After the treatment of two groups of volunteers,all the volunteers after the end of treatment did not appear scar and skin hypopigmentation phenomenon.After treatment,the pain score,erythema duration and swelling duration of the observation group were significantly better than those of the control group,with the difference being statistically significant(P < 0.05).However,there was no significant difference between the two groups in epidermal callus removal duration(P > 0.05).The above results show that the improvement rate of 800 nm semiconductor laser treatment of nasal blackheads and coarse pores was better than Q-switched Nd:YAG laser.The incidence of adverse reactions was lower than that of Q-switched Nd:YAG laser treatment.Conclusion:1 Light Sheer Duet 800 nm semiconductor laser therapy can effectively reduce the number and volume of hair follicles and sebaceous glands of the black skin guinea pig,and does not lead to damage dermal elastic fiber fracture,Light Sheer Duet 800 nm high-dose semiconductor laser treatment group was better than that of the low-dose group and Q-switched laser treatment group.2 Light Sheer Duet 800 nm semiconductor laser can promote the apoptosis of black guinea-pig skin sebaceous gland cells,through up-regulation expression of Bax protein and down-regulation of Bcl-2 protein,decreasing Bcl-2/Bax ratio,to up-regulate the activation of caspase-3.3 The improvement rate of 800 nm semiconductor laser treatment of nasal blackheads and coarse pores was better than Q-switched Nd:YAG laser.The incidence of adverse reactions was lower than that of Q-switched Nd:YAGlaser treatment. |
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