Objective To investigate the distribution of Malassezia species in lesional and non-lesion-al sites of patients with pityriasis versicolor(PV),species-variation in different anatomic sites and in lesions with different pigmentation,and the relationship between various Malassezia species and severity and age of PV patients.Methods A total of629skin specimens taken by sterile adhesive tape from the lesions and non-lesional skin were inoculated on media containing rapeseed oil in113patients with PV.Isolated colonies were identified to species based on physiological and morphological characteristics.Results The isolation rates of Malassezia spp.were not significantly different from both lesions and corresponding non-lesional skin.Among non-lesional sites,the isolation rate was significantly higher in forehead and trunk than that in upper and lower extremities.Five species were identified out of565strains obtained from the patients,including M.sympodialis(44.78%),M.furfur(32.94%),M.globosa(11.68%),M.obtusa(5.84%)and M.restricta(4.76%).Two dif-ferent species were isolated simultaneously from27sites.There was no obvious difference in species distribu-tion patterns between lesions and non-lesional sites.M.restricta was isolated from forehead exclusively.Species-variation was closely linked to lesions with different pigmentation and the age of patients,not to the severity of disease.Conclusion There is neither statistical difference of Malassezia isolation rate and species distribution between lesions and non-lesional skin,nor correlation between disease severity and species-varia-tion.The anatomic sites,the diversity of pigmentation pattern and the age of patients seem to be associated with different Malassezia species.

Objective To study on the association of apolipoprotein E(apoE)genotype with coronary heart disease(CHD)and type 2 diabetes mellitus(T2DM).Methods PCR-RFL,multiplex amplification refractory mutation system(muli-ARMS)and PCR-SSCP methods were used to detect the genotype of apoE,and DNA sequencing technique were used for further confrm the genotype and gene variations in 2 446 Chinese individuals,including 238 cases of CHD,316 cases of T2DM and 1 892 healthy controls.Fasting blood glucose(FBG)and plasma lipids levels[TC,TG,HDL-C,LDL-C,apoA I,apoB and Lo(a)]were measured by usual methods.Results Compared with the controls,plasma HDL-C(t=2.66)and apoA I(t=2.30)levels in the CHD group were significantly lower(P<0.05),but not in T2DM group;plasma TC level(t=5.22)in the T2DM group were significantly higher(P<0.05),but not in CHD group;systolic pressure(t=8.48,5.74)diastolic pressure(t=5.66,3.35),plasma TG(t=3.38, 4.56),LDL-C(t=2.48,7.00),apoB(t=1.67,2.24),Lp(a)(t=4.16,4.15)and FBG(t=7.04, 16.93)levels were significantly higher in both CHD group and T2DM group(P<0.05).The distributions of apoE ε2/2,ε2/3,ε3/3,ε2/4,ε3/4 and ε4/4 respectively were 0.4%,13.4%,58.0%,1.3%, 26.5%,0.4% in the CHD group;0.6%,5.7%,72.8%,1.9%,14.9%,4.1% in the T2DM group; 0.5%,10.5%,69.6%,1.6%,16.8%,1.1% in the control group.Significant differences were found between the CHD group(χ2=14.90,P=0.00),T2DM group(χ2=7.08,P=0.03)and the control group for the frequencies of apoE genotype.The distribution of ε3/4 was higher(26.5% vs 16.8%)and ε3/3 Was lower(58.0% vs 69.6%)in the CHD group.In the T2DM group.the distribution of εε4/4 Was higher (4.1% vs 1.1%),and 2 cases of ε3/3 with Arg 150 His mutation in exon 4 of apoE gene were firstly reposed in China,which is none in the CHD and control groups.Conclusions The results suggested that apoE ε3/4 and ε4 genotypes might be associated with the susceptibility of CHD and T2DM.respectively. To some extent,apoE ε3/3 may not be a good genotype for T2DM because of the Arg 150 His mutation. Blood pressure and plasma lipids could be used for diagnosis of the two diseases.

Objective To investigate changes in expressions of activation antigens CD69 and HLA-DR in CD3+T lymphocytes in peripheral blood and skin lesions in patients with psoriasis vulgaris. Methods Peripheral blood samples were obtained from 20 patients with psoriasis vulgaris and 20 healthy controls, and skin specimens from the lesions of 15 out of the 20 patients and 10 healthy controls. Flow cytometry was performed to quantify the expressions of CD69 and HLA-DR in peripheral blood CD3+T cells, and an immunohistochemical study to measure the expression of HLA-DR in skin specimens. Statistical analysis was carried out by a two-sample t-test and Pearson correlation analysis with the SPSS 19.0 software. Results Compared with the healthy controls, the patients with psoriasis vulgaris showed increased expression rates of CD69 (4.70%± 1.90%vs. 1.56%± 0.95%, t=6.629, P<0.01)and HLA-DR (8.97%± 1.79% vs. 3.02% ± 1.15%, t= 6.204, P< 0.01)in peripheral blood. Pearson correlation analysis revealed that the percentage of CD3+HLA-DR+cells in peripheral blood was positively correlated with the psoriasis area and severity index (PASI)score (r=0.5626, P<0.05). The expression rate of HLA-DR was significantly higher in the dermis (64.87%± 17.31%vs. 19.80%± 5.69%, t=7.916, P<0.01), but lower in the epidermis(11.80%± 5.55%vs. 27.40%± 8.61%, t=5.479, P<0.01)in the psoriatic specimens compared with the control specimens. Immunohistochemically, HLA-DR was widely expressed in the dermis of psoriatic lesions, but mainly distributed around blood vessels in the control skin. Conclusions There is an aberrant activation of CD3+T cells in peripheral blood and inflammatory cells in skin lesions in patients with psoriasis vulgaris, and the percentage of CD3 +HLA-DR+ cells in peripheral blood is correlated with the severity of psoriasis vulagaris.

Objective To report a case of disseminated cryptococcosis with cutaneous manifestations and osteomyelitis. Methods and Results A 33 year old female was admitted due to multiple nodules and ulcers on the upper arms, shoulders, buttocks and thighs for one year. The patient was pregnant when admitted, and gave birth to a premature baby during her illness. The nodules increased half a month after delivery, which was suspected to be hematogenously disseminated pulmonary tuberculosis and was given anti tuberculous therapy for three months but failed. Physical examination showed there were 39 nodules or ulcers on the face, gum, trunk, buttocks and extre mities. The bone structure of the left tibia and fibula destroyed and a sinus developed on the left fibula. Microbiologic examination showed that lots of spores were seen in the smear of pus and necrotic tissues, which produced yeast like colonies in culture with positive urease and caffeic acid test. Cryptococcus neoformans, serotype A was identified by API yeast reaction band and serology. Inoculation with mice and rats showed that their brains, lungs and livers were involved easily. Further identification as C.neoformans var.neoformans was obtained based on sequence analysis of ribosomal internal transcribed spacer region 2. The anti tuberculous therapy was stopped and anti fungal therapy was initiated at once. Intravenous and topical amphotericin B in combination with fluconazole were chosen in the initial therapy and itraconazole for maintenance. The nodules disappeared after 30 days and the last ulcer in the left tibia healed completely after 200 days. The anti fungal therapy was discontinued after 277 days and the patient was completely cured.

To strengthen the quality control of palbociclib, three related substances were separated from the bulk drug. These substances were identified as 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)pyrido ［2, 3-d］ pyrimidin-7(8H)-one(A), 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridine-2-yl)amino)-6-vinylpyrido ［2, 3-d］ pyrimidin-7(8H)-one(B), tert-butyl4-(6-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7, 8-dihydropyrido ［2, 3-d］ pyrimidin-2-yl)amino)pyridin-3-yl)piperazi-ne-1-carboxylate(C). Based on the structures of the impurities, the possible routes to them were discussed, and their structures were elucidated by 1H NMR and MS.

Objective:To investigate the surgical method and clinical effect of free medial plantar flap (MPF) combined with anterolateral thigh flap (ALTF) in repair of large soft tissue defect in the weight bearing area of sole.Methods:From April, 2017 to August, 2019, 8 patients with large soft tissue defects in plantar weight bearing area were repaired by free MPF combined with ALTF. Four patients had the defects located in the hindfoot and the surrounding area, 3 in the forefoot and 1 in the whole foot. A tandem flap made of free MPF combined with ALTF was used to repair the heel in 5 patients and repair the plantar forefoot in 3 patients. The size of defects ranged from 15.0 cm×10.0 cm to 26.0 cm×22.0 cm. The size of the MPF ranged from 6.0 cm×5.0 cm to 8.0 cm×6.5 cm. The donor sites repaired with full thickness skin graft. The size of ALTF ranged from 15.0 cm×7.0 cm to 27.0 cm×11.0 cm. Two donor sites were sutured directly, and the other 6 were repaired by full-thickness abdominal skin graft. The patients entered follow-up at outpatient clinic and via WeChat for 9 to 18 months.Results:All the 8 tandem flaps and the donor grafts survived. Only 1 ALTF had a distal necrosis and healed after change of dressing. All the flaps had good elasticity and good texture. All the recipient area of MPF achieved sensation recovery of pain and touch. But the ALTF only partially recovered tactile sensation. The weight-bearing and walking function were good. At the last follow-up, all patients were evaluated by Maryland foot score, of which 4 patients were excellent, 3 were good, and 1 was fair.Conclusion:The free MPF combined with ALTF is one of the ideal methods for the repair of plantar soft tissue defect in the weight bearing area of sole. It can better restore the foot weight-bearing and walking function with good clinical effect.

Objective To explore the clinical effect of repairing the large area of soft tissue defect of the calf by the retrograde anterolateral thigh flap with single high cutaneous perforator. Methods From January, 2014 to July, 2017, 9 cases of large area of soft tissue defects were repaired by the retrograde anterolateral thigh flap with sin-gle high cutaneous perforator.There were 7 males and 2 females, aged 24-48 years.Soft tissue defects area of the calf was 10.0 cm×7.0 cm to 35.0 cm×15.0 cm, including skin grafting and skin stretch to repair the area. The perforating point of the high cutaneous artery branches was designed at the proximal end of the flap, which was used as the single nutrient vessel of the flap. The rotation point of the flap was moved upward to the proximal thigh, which not only in-creased the blood supply of the flap, but also made the flap repair range to the distal calf. The flap range was 15.0 cm×10.0 cm to 22.0 cm×12.0 cm. Results All flaps were cut smoothly, and no vascular crisis occurred. All flaps survived smoothly.All patients were followed-up for 6-12 months. The appearance of flaps was plump, slightly bloat-ed, and their color was similar to the recipient area. The texture was soft, and no active disorder in the donor site. Conclusion The retrograde anterolateral thigh flap with single high cutaneous perforator can be designed at a high rotation point.By increasing the number and caliber of the anastomotic branch between the pedicle and lateral superi-or genicular artery, the blood supply and reflux of flap can be improved, and the survival rate is not affected. Com-pared with the traditional anterolateral thigh flap, it has great advantages.

Objective To investigate the clinical effect of lateral supramalleolar perforator flap with the rotation point down on the repair of forefoot soft tissue defect.Methods From January,2016 to June,2017,23 cases (15 males and 8 females) of forefoot soft tissue defect were repaired by lateral supramalleolar perforator flap with the rotation point down.The age ranged from 8 to 67 (mean,42.9) years.The soft tissue defects were located far from the tarsometatarsal joint of the foot and the areas were 4.0 cm×4.0 cm-8.0 cm×12.0 cm.The areas of flap stitched directly or take blade thick skin repair on the thigh.All cases were followed-up by telephone or Internet,outpatient visit and home visit.Results Twenty-one cases survived successfully.Small area necrosis occurred at the distal end of the flap in 1 case,and healed after changing treatment.In the other cases,tension blister appeared at the distal end of the flap.After cut off some surgical stitches,the flap survived.All cases were followed-up from 6 to 12 months.The shape of flaps were not bloated and had good textures.The walking of affected limbs had no significant effect,and the functional recovery was satisfactory.Conclusion The lateral supramalleolar perforator flap is easy to cut and the perforator of flap is constant,which is a appropriate flap to repair the soft tissue defect of the forefoot.

To explore the method and effect of repairing the soft tissue defect of the lateral heel with the retrograde lateral supramalleolar flap pedicled with the end perforator of peroneal artery. Methods From May, 2015 to February, 2018, 16 cases of lateral calcaneal soft tissue defect were repaired with the retrograde lateral supramalleolar flap pedicled with the end perforator of peroneal artery.All wounds were treated with one-stage dilata-tion and VSD to control infection. In cases of chronic calcaneal bone infection, the bone defect formed after extensive resection of infected bone was temporarily filled with antibiotic bone cement. The area of soft tissue defect on the lat-eral heel was 3.0 cm×2.0 cm-8.0 cm×5.0 cm, and the area of flaps was 3.5 cm×2.5 cm-8.5 cm×5.5 cm. The small donor area of the flap was sutured directly, and the larger area was repaired by skin grafting. Patients with chronic calcaneal bone infection underwent bone cement removal and autogenous bone transplantation after inducing mem-brane formation 6 to 8 weeks after flap transplantation. All cases were followed-up, including 7 cases outpatient fol-low-up and 9 telephone follow-up. Results All the 16 flaps survived smoothly. The donor and recipient areas of the flaps healed primarily. All cases were followed-up for 3 to 13 months. The flaps had good shape, no swelling, similar color to heel skin and no pigmentation.Ankle flexion and extension were not restricted.Four cases with chron-ic osteomyelitis of calcaneus healed well after second-stage bone grafting, with an average healing time of 8.5 months. Conclusion The retrograde lateral supramalleolar flap with the end perforator of peroneal artery is an ideal method for repairing the soft tissue defect on the lateral heel with simple operation and reliable blood supply.

Objective:To explore a surgical technique and treatment outcomes of the segmentle bridging reconstruction for severed fingers with single segmentle defect by using the free second toe bone-joint composite tissue combined with the great toe flap.Methods:From June 2010 to September 2017, 5 patients suffered from severed segmental defect of finger were treated. According to the defects of bone-joints, blood vessels, nerves, tendons and other soft tissues, the reconstruction surgery was designed to create a Flow-through bridging composite flap pedicled with the first dorsal metatarsal artery or the plantar artery. The blood vessles carried by the pedicle were anastomosed with the vessels in the finger to restore the blood supply to the distal finger while having the defected finger segment reconstructed. A Flow-through bridging composite flap was created by taking the second toe bone-joint composite tissue combined with a C-shaped or half-moon shaped flap from the fibular side of the great toe. Skin graft, retrograde lateral tarsal flap or free perforator flaps were used in 3 cases to repair the donor sites of the great and second toes. Iliac strip was implanted in 2 cases for toe salvage. Kirschner wires were removed 4-6 weeks after surgery followed by functional exercise.Results:All of the 5 reconstructed distal segments of the fingers survived with the healing of fractures in 8-12 week after surgery. The postoperative follow-up lasted 6-36 months and all the patients had the follow-ups at the outpatient clinic. It was found that the maximum flexion of the reconstructed interphalangel joint was 60 degrees together with dorsiflezion. According to the evaluation standard of the reconstructed function for thumb and finger issued by the Hand Surgery Society of the Chinese Medical Association, 3 fingers were in excellent and 2 in good. No obviouse affect on walking was found in all the patients.Conclusion:The free second toe joint composite tissue together with the great toe flap can be used to bridge the single segmental defect of a finger. It restores the blood supply to as well as the appearance of the distal finger, helps the recovery of the reconstructed distal finger. It is an ideal technique in the treatment of a severed distal segment of the finger.