Objective To investigate the effect of heat treatment on the proliferation of, melanin synthesis and tyrosinase activity in cultured normal human melanocytes. Methods Normal human foreskin tissue was obtained by sterile circumcision and melanocytes were harvested by using methods for epidermal cell culture. Basic fibroblast growth factor (bFGF) was utilized as the primary mitogen to establish the culture system of normal human epidermal melanocytes. Masson-Fontana staining was proformed to identify melanocytes.Third-passage melanocytes were treated with hyperthermia at various temperatures (39 ℃, 41 ℃, 42 ℃, 43 ℃ and 45℃) for 1 hour a day for consecutive 3 days followed by the measurement of cell viability with MTT assay. The hyperthermia at optimized temperature was used to treat fourth-passage melanocytes for 1 hour a day for consecutive 3 days; subsequently, the tyrosinase activity were detected with L-Dopa as the substrate, and melanin content was determined in heat-treated and untreated (control) melanocytes. Results The hyperthermia at 42 ℃ exhibited the strongest promotive effect on the proliferation of melanocytes among these 5 hyperthermia conditions. After treatment with hyperthermia at 42 ℃ for 1 hour a day for consecutive 3 days, melanocytes showed an increment in tyrosinase activity by 36.4% and melanin synthesis by 78% compared with the untreated melanocytes (both P＜0.05). Conclusions Heat treatment can enhance the proliferation of cultured human melanocytes, promote their melanin synthesis and tyrosinase activity.

BACKGROUND:Intra-articular injection played an important role in the treatment of osteoarthritis and has more options with the development of novel drug delivery systems.The cartilage targeting function is aimed at the adhesion or retention of drugs in the cartilage layer to form a drug bank to achieve slow release and precise drug delivery. OBJECTIVE:To review various cartilage targeting biomaterials and their characteristics in the treatment of osteoarthritis by articular injection. METHODS:Using the term"osteoarthritis,drug carrier,drug delivery,cartilage targeting,penetrate"as key words,relevant articles were searched in CNKI,WanFang and PubMed databases.According to inclusion and exclusion criteria,67 articles were finally selected for further review. RESULTS AND CONCLUSION:The research on cartilage-targeting biomaterials is mainly divided into two directions.One is the combination of electrostatic interaction,such as the combination of positively charged biomaterials and negatively charged polysaccharides in cartilage.This kind of scheme is operable and easy to modify,but limited by the shortcomings of electrostatic interaction itself,it performs badly in advanced osteoarthritis.Another one is the specific binding of various components in cartilage which is strong and reliable,and related biomaterials have excellent performance in advanced osteoarthritis,which is an important direction for future cartilage-targeted therapy.

Objective: To investigate the expression levels of S100A6, Notch1 in multiple myeloma (MM) patients and its clinical significance. Mathods: The expression levels of S100A6, Notch1 in 28 MM cases and 20 healthy controls were determined by real time quantitative PCR (RQ-PCR) , and their relationships with clinical features and outcomes were analyzed. Immunohistochemical was used to analysis the levels of S100A6 and Notch1 in bone marrow biopsy samples and intramedullary metastases soft tissues. RQ-PCR and Western blot were used to test the changes of Notch1 mRNA and Notch1 protein in U266 MM cells after S100A6 silenced by siRNA. Results: ①The expression levels of S100A6, Notch1 in primary MM patients was 2.19±1.25, 2.98±0.64, significantly higher than those in controls (0.71±0.20, 0.58±0.39, P<0.05) and patients in platform status (0.85±0.26, 0.72±0.40, P<0.05) . 8 cases with intramedullary metastasis had significantly higher levels of S100A6 (3.36±1.23) and Notch1 (5.71±3.96) , as compared to those without extra medullary metastases. ②S100A6 expression was positive correlation with Notch1 (r=0.505, P=0.007) . ③S100A6 and Notch1 proteins were positive in plasma cells of bone marrow biopsy samples and intramedullary metastases soft tissues. ④The Notch1 mRNA and Notch1 expression decreased significantly after 48 hours treatment by S100A6 siRNA in U266 cells. Conclusion S100A6 and Notch1 were closely associated with MM progress and intramedullary metastasis. They have significant correlation and might be as two prognostic molecular markers in MM.

Objective To investigate the effect of the peroxide proliferator-activated receptorgamma (PPAR-γ) agonist rosiglitazone on the motor function recovery of hind limbs in rats with spinal cord injury.Methods Sixty-eight female SD rats were used to establish spinal cord injury model by modified Allen method.(1) Eight rats were randomly divided into negative control group and rosiglitazone group with four rats in each group.The expression of aspartate proteolytic enzyme-1 (caspase-1) in spinal cord of rats 7 days after injury was detected by immunohistochemical staining.(2) Forty-eight rats were randomly divided into negative control group,rosiglitazone group,rosiglitazone + Clostridium chitosans group [nuclear factor kappa B (NF-kappa B) activator],rosiglitazone + monosodium urate group [oligomerization domain-like receptor protein 3 (NLRP3) antagonist],with 12 rats in each group.BBB scores of hindlimb motor function were assessed at 1,3,14,21 and 28 days after injury in each group.The expression of interleukin-1 β (IL-1 β) and tumor necrosis factor-α (TNF-α) in each group was detected by ELISA at 28 days after injury.Microglia were isolated from the spinal cord of 12 rats and cultured for 7 days.They were randomly divided into the following five groups:(1) negative control group:no drug treatment;(2) rosiglitazone group:1 micromol/L rosiglitazone treatment;(3) rosiglitazone + Clostridium chitin group:1 micromol/L rosiglitazone + 20 micromol/L Clostridium chitosporin treatment;(4) Clostridium chitosan treatment Mycoplasma group:20 μ mol/L shell Clostridium treatment;(5) Clostridium chitosanin + MCC950 group [(NLRP3) antagonist]:20 μmol/L Clostridium chitosanin + 100 nmol/L MCC950;Western blot was used to detect the expressions of caspase-1,NF-kappa B and NLRP3 in microglia cells;ELISA was used to detect the expressions of IL-1β and TNF-α in the supernatant of microglia culture.Results Compared with negative control group,caspase-1 expression was decreased in rosiglitazone group in spinal cord injury area [gray matter area:5.1 ± 0.8∶6.9 ± 1.1;white matter area:5.6 ± 0.9 ∶ 7.5 ± 1.1] (P ＜ 0.05).At 28 days after operation,the rosiglitazone group had the highest BBB score [(14.7 ± 1.6) points],and the BBB score of rosiglitazone + Clostridium chitosans group (10.5 ± 2.1) points was superior to that of rosiglitazone + monosodium urate group [(7.2 ± 1.3)points,P ＜ 0.05].The expressions of IL-1β and TNF-α in rosiglitazone + monosodium uric acid group were lower than those in other groups at 28 days after injury (P ＜ 0.05).In vitro,the expressions of caspase-1,NF-kappa B,IL-1β and TNF-α in rosiglitazone group were lower than those in negative control group (P ＜ O.05).The expressions of caspase-1,NLRP3,IL-1β and TNF-α in rosiglitazone + Clostridium chitosani group were higher than those in rosiglitazone group,(P ＜ 0.05).The expressions of caspase-1 and IL-1β were higher than those in Clostridium chitosani + MCC950 group (P ＜0.05),but there was no significant difference in the expression of TNF-α between the two groups (P ＞0.05).Conclusion Rosiglitazone can promote the recovery of hind limb motor function in rats with spinal cord injury by inhibiting the expression of NF-kappa B,thereby reducing the activation of NLRP3 inflammatory bodies in microglia and ultimately inhibiting the occurrence of inflammation.

Objective:To evaluate the patch technique in repairing huge rotator cuff tear.Methods:A retrospective analysis was conducted of the 9 patients with huge rotator cuff tear who had been repaired with patch technique at Department of Orthopaedics, Guangzhou Red Cross Hospital from March 2017 to March 2019. They were 5 males and 4 females, aged from 53 to 79 years (average, 61 years). Shoulder movement limitation was found in 7 cases, night pain in 5, and positive Neer impingement sign and Hawkins sign in 7. By the Bigliani acromion classification, there were 6 cases of type Ⅱ and 3 cases of type Ⅲ. Comparisons were made between preoperation, 12 and 15 months postoperation in terms of scores of the visual analogue scale(VAS) and the University of California Los Angeles (UCLA) scoring system, and shoulder range of motion.Results:All the 9 patients were followed up for 15 to 24 months(mean, 18 months). Arthroscopy found tears in more than 2 tendons in all of them. The VAS scores were 6.7±1.6, 4.5±1.3 and 3.7±1.1 at preoperation, 12 and 15 months postoperation; the UCLA scores were 7.9±1.2, 21.5±4.1 and 23.9±4.3 at preoperation, 12 and 15 months postoperation. There were statistically significant differences in both the VAS and UCLA scores between preoperation, 12 and 15 months postoperation ( P<0.05). There were also statistically significant differences between the 3 groups in the shoulder range of motion at preoperation, 12 and 15 months ( P<0.05). MRI reexamination at 12 months postoperation showed minor re-tear < 3 cm in 2 patients. Conclusion:The patch technique is a reasonable and effective treatment to repair huge rotator cuff tears, resulting in good mid-term outcomes.

OBJECTIVE: To detect potential mutations of the PKHD1 gene in two pedigrees affected with infantile polycystic kidney disease. METHODS: Clinical data and peripheral venous blood samples were collected from the probands and their parents as well as fetal amniotic fluid cells. Genome DNA was extracted from the peripheral blood samples and amniotic fluid cells. Exons 32 and 61 of the PKHD1 gene were amplified with PCR and subjected to direct sequencing. RESULTS: The proband of pedigree 1 was found to carry c.4274T>G (p.Leu1425Arg) mutation in exon 32 and c.10445G>C (p.Arg3482Pro) mutation in exon 61 of the PKHD1 gene, which were inherited from her father and mother, respectively. The fetus has carried the c.4274T>G (p.Leu1425Arg) mutation. In pedigree 2, the wife and her husband had respectively carried a heterozygous c.5979_5981delTGG mutation and a c.9455delA mutation of the PKHD1 gene. No chromosomal aberration was found in the umbilical blood sample, but the genetic testing of their fetus was failed. Based on software prediction, all of the 4 mutations were predicted to be pathogenic. CONCLUSION PKHD1 c.4274T>G (p.Leu1425Arg), c.10445G>C (p.Arg3482Pro), c.5979_5981delTGG and c.9455delA were likely to be pathogenic mutations. The results have facilitated genetic counseling and prenatal diagnosis for the two pedigrees.
DNA Mutational Analysis ; Female ; Genetic Counseling ; Humans ; Mutation ; Pedigree ; Polycystic Kidney Diseases ; diagnosis ; genetics ; Pregnancy ; Prenatal Diagnosis ; Receptors, Cell Surface ; drug effects

Objective:To investigate the reparative efficacy and mechanism of pressure-adjustable macroporous antibacterial hydrogel in the treatment of infected wounds.Methods:Staphylococcus aureus was used to establish wound infection models in healthy C57BL/6 mice. The models were divided into 3 groups subjected to 3 different treatments: a negative control group with no hydrogel treatment (group A), a control group treated by common medical hydrogel (group B) and an experiment group treated by pressure-adjustable macroporous antibacterial hydrogel (group C). On days 1, 3, 6, 9 and 12, the effects of 3 treatments were compared on the wound area and the number of bacterial colonies under scab, on the apoptosis of fibroblasts based on the changes of type Ⅰ procollagen, and on the inhibition of inflammation during wound repair by detecting the expression of interleukin-6 (IL-6) and tumor necrosis factor (TNF-α).Results:On days 1 and 3, there was no significant difference between the 3 groups in the wound area ( P>0.05), but on days 6, 9 and 12, there were significant differences between the 3 groups in the wound area ( P<0.05). On day 6, the wound areas in group B (1.23 cm 2 ± 0.16 cm 2) and in group C (1.14 cm 2 ± 0.12 cm 2) were significantly smaller than that in group A (1.56 cm 2 ± 0.16 cm 2) ( P<0.05), but there was no significant difference between groups B and C ( P>0.05). On days 9 and 12, the wound areas in group B (0.97 cm 2 ± 0.13 cm 2 and 0.76 cm 2 ± 0.10 cm 2) and in group C (0.66 cm 2 ± 0.06 cm 2 and 0.48 cm 2 ± 0.07 cm 2) were significantly smaller than those in group A (1.49 cm 2 ± 0.11 cm 2 and 1.39 cm 2 ± 0.13 cm 2), and those in group C were significantly smaller than those in group B (all P<0.05). On day 1, there was no significant difference between the 3 groups in the number of bacterial colonies under scab ( P>0.05). On days 3, 6, 9 and 12, the numbers of bacterial colonies under scab in groups B and C were significantly smaller than that in group A ( P<0.05), and that in group C was significantly smaller than that in group B ( P< 0.05). The nucleic acid electrophoresis showed that the grayscale bands in group C were significantly darker than those in groups A and B. The early apoptosis rate of the fibroblasts in group C[low-right positive fluorescence (LR%): 9.72%] was significantly lower than that in group A (43.99%) and that in group B (38.43%), and that in group B was significantly lower than that in group A ( P<0.05). On day 12, the ratio of the gray values of IL-6 and β-actin (0.64 ± 0.10) and the ratio of the gray values of TNF-α and β-actin (0.34 ± 0.05) in the fibroblasts in group C were significantly higher than those in group A (1.22 ± 0.21 and 0.60 ± 0.14) and in group B (0.88 ± 0.02 and 0.41 ± 0.06) ( P<0.01). Conclusion:The pressure-adjustable macroporous antibacterial hydrogel is an effective treatment of infected wounds and its mechanism may be related to the reduced apoptosis of fibroblasts.

Objective:To improve the understanding of indolent mantle cell lymphoma (MCL).Methods:The data of a patient with indolent leukemic MCL in the Second Affiliated Hospital of Nanjing Medical University in May 2013 were collected. The cell morphology was analyzed by using cell smear, the flow cytometry was used to make immunophenotype analysis, the karyotype analysis was performed by usig cytogenetic technique, and polymerase chain reaction (PCR) was used to make the immunoglobulin gene analysis. At the same time, lymph node pathology and immunohistochemistry were also analyzed. The related articles published were reviewed to sum up the characteristics and the treatment of indolent MCL.Results:The male patient aged 60 years was obviously asymptomatic accompanied with slow disease progression, leukemic manifestation and without lymphadenopathy. He received pathological biopsy because of located lymphadenopathy in 2008. Small cell morphology, Kappa light chain immunophenotype, t(11;14) translocation showed after the cytogenetic examination, clonal immune globulin gene rearrangement and low Ki-67 positive index were identified. In situ MCL was diagnosed by retrospective pathology.Conclusions:Indolent MCL is extremely rare. It is typically asymptomatic with none or minimal nodal involvement, indolent disease course, leukemic phase with mild lymphocytosis, Kappa light chain expression, simple karyotype, classical or small cell morphology of tumor cells and the positive index of Ki-67 <10%. In situ MCL can be seen in pathology examination. IgVH gene mutation positive and SOX11 negative expression are notable in indolent MCL. International prognostic index of MCL is probably not appropriate in the prognostic analysis of leukemic indolent MCL. It is emphasized that initial observation and having therapies only after the disease progression can be suited for indolent MCL.

OBJECTIVE: To investigate the chemical constituents from the leaves of Jatropha curcas and evaluate their inhibition on lipopolysaccharide (LPS)-activated BV-2 microglia cells. METHODS: The n-BuOH extract of the leaves of J. curcas was isolated by macroporous adsorption resin, silica gel, ODS, column chromatography and semi-preparative HPLC. The structures of the compounds were identified by MS, NMR, ECD, and other spectroscopic methods. In addition, anti-neuroinflammatory effects of isolated compounds were evaluated by measuring the production of nitric oxide (NO) in over-activated BV-2 cells. RESULTS: Seventeen compounds, including (7R,8S)-crataegifin A-4-O-β-D-glucopyranoside ( 1), (8R,8'R)-arctigenin ( 2), arctigenin-4'-O-β-D-glucopyranoside ( 3), (-)-syringaresinol ( 4), syringaresinol-4'-O-β-D-glucopyranoside ( 5), (-)-pinoresinol ( 6), pinoresinol-4'-O-β-D-glucopyranoside ( 7), buddlenol D ( 8), (2R,3R)-dihydroquercetin ( 9), (2S,3S)-epicatechin ( 10), (2R,3S)-catechin ( 11), isovitexin ( 12), naringenin-7-O-β-D-glucopyranoside ( 13), chamaejasmin ( 14), neochamaejasmin B ( 15), isoneochamaejasmin A ( 16), and tomentin-5-O-β-D-glucopyranoside ( 17) were isolated and identified. Compounds 2, 4 and 8 significantly inhibited the release of NO in BV-2 microglia activated by LPS, with IC₅₀ values of 18.34, 29.33 and 26.30 μmol/L, respectively. CONCLUSION Compound 1 is a novel compound, and compounds 2, 3, 8, 14- 17 are isolated from Jatropha genus for the first time. In addition, the lignans significantly inhibited NO release and the inhibitory activity was decreased after glycosylation.