Objective To explore the inhibition and apoptosis of the human cutaneous T-cell lymphoma cell lines Hut-78 by traditional Chinese medicine SVCⅢ. Methods After Hut-78 cells were treated with SVCⅢ of different concentration, the inhibition and apoptosis of Hut-78 cells was determined by MTT, agarose gel electrophoresis of DNA fragment and FACS. Results SVCⅢ could inhibit remarkably Hut-78 cells growth and DNA ladder was seen by agarose gel electrophoresis. The proliferation of Hut-78 cells were inhibited in G_1 stage by FACS. Conclusion SVCⅢ can promote growth retardation and apoptosis of human cutaneous T-cell lymphoma cell lines Hut-78, which suggests SVCⅢ has antineoplastic function.

Objective To construct eukaryotic cell expression vector of human frangible histone triad (FHIT) gene. Methods A 456 bp cDNA fragment was amplified from the total RNA of normal human thyroid tissue by RT PCR method and cloned into plasmid pcDNA3. The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes Kpn Ⅰ and Bst XⅠ and sequenced by Sanger dideoxy mediated chain termination. The expression of FHIT gene was detected by immunocytochemical methods. Results The results showed that the cDNA fragment included 456 bp entire coding region. The recombinant eukaryotic cell expression vector of pcDNA3 FHIT was constructed, and the sequence of the insert was identical to the published sequence. MM96L cells transfected with the pcDNA3 FHIT plasmid expressed high level of Fhit protein in cytoplasm. Conclusion The recombinant plasmid pcDNA3 FHIT can provide a strong molecular tool for the studies of neoplasm pathogenesis.

Objective To detect the FHIT gene promoter methylation and protein expression in mycosis fungoides(MF).Methods Tissue specimens were collected from 48 patients with MF and 18 normal human controls.FHIT protein expression was determined by immunohistochemistry,and methylation status of FHIT gene by methylation-specific PCR.Results Abnormal methylation of FHIT gene was found in 26(54.2%)out of the 48 specimens.Thirty(63.5%)specimens of MF were negative for FHIT protein,which was observed in all the control specimens.The promoter methylation of FHIT was closely correlated with the protein expression of FHIT,but unrelated to the sex of,tumor staging or lymph node metastasis in patients with MF.Conclusion The FHIT gene promoter methylation may contribute to the inactivation and abnormal expression of FHIT protein in MF.

Objective To evaluate the protective effect of epigallocatechin gallate (EGCG) against interleukin (IL)-17-induced injury to keratinocytes,and to explore its mechanism.Methods Some cultured HaCaT cells were divided into 3 groups to be treated with IL-17 alone at concentrations of 50,70,90 μg/L,respectively,with those receiving no treatment as the blank control group.Some HaCaT cells were divided into 5 groups:IL-17 group treated with 90 μg/L IL-17 alone,IL-17 + EGCG group treated with 90 μg/L IL-17 and 60 μmol/L EGCG,IL-17 + SP600125 group treated with 90 μg/L IL-17 and SP600125 (a JAK signaling pathway inhibitor),IL-17+ EGCG + anisomycin group treated with 90μg/L IL-17,60xmol/L EGCG and anisomycin (a Janus kinase signaling pathway activator),and blank control group receiving no treatment.After different durations of treatment,CCK-8 assay was performed to evaluate cellular proliferative activity,flow cytometry to detect cell apoptosis,enzyme-linked immunosorbent assay (ELISA) to measure expression levels of IL-6,IL-23 and IL-8,and Western-blot analysis to determine protein expressions of c-Jun N-terminal kinase (JNK) and phosphorylated JNK (P-JNK).Results IL-17 promoted cellular proliferation of HaCaT cells,and the proliferation rate,which was correlated with the concentration of IL-17,reached the maximum in the 90-μg/L IL-17 group (P ＜ 0.05).EGCG at 60 μmol/L significantly inhibited cellular proliferation of,promoted apoptosis in,and reduced IL-6,IL-23 and IL-8 expressions in,HaCaT cells induced by 90 μg/L IL-17 (all P ＜ 0.05).Compared with the IL-17 group,the IL-17 + EGCG group and IL-17 + SP600125 group both showed significantly decreased P-JNK expression,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).However,compared with the IL-17 + EGCG group,the IL-17 + EGCG + anisomycin group showed significantly increased protein expression of P-JNK,cell proliferation rate and IL-6,IL-23 and IL-8 expression levels (all P ＜ 0.05).Conclusion EGCG protected against IL-17-induced injury to HaCaT cells,such as abnormal cell proliferation,apoptosis and inflammatory response,likely by inhibiting the JNK signaling pathway.

Objective To investigate the effect of the exogenous fragile hisdidine triad(FHIT) gene on the proliferation and the apoptosis of cutaneous carcinoma cell line A431,and to explore the mechanism of tumor suppression by the FHIT gene.Methods The plasmids pcDNA3-FHIT and pcDNA3-vector were transfected into the cutaneous carcinoma cell line A431 without FHIT gene expression,and then the transfected cells were screened by G418 and the expression of FHIT was determined by the immunocytochemical staining technique.The effect of FHIT on the growth characteristics of cutaneous carcinoma cell line A431 was observed by MTT,colony forming test and flow cytometry.Results Stable FHIT gene expressing A431 cells were produced,the proliferation activity and colony forming capability of A431FHIT were suppressed,whereas the apoptosis was increased.All these differences between A431-FHIT cells and the two control groups of cutaneous carcinoma cells had statistical significance.Conclusion Transfecting the exogenous FHIT gene into cutaneous carcinoma cells line A431can suppress the proliferation of tumor cells,and can also induce apoptosis and cell cycle arrest.

The exprimental teaching of medical immunology is the important constituent.We have explored the existing problems on course content,teaching method and experimental test way according to our practical teaching experience for the past few years.We have also made the preliminary attempt about reforming exprimental teaching of medical immunology.

Objective:To evaluate the curative effects of Masquelet technique and 3D printing in repair of Cierny-Mader type Ⅳ long bone osteomyelitis.Methods:A retrospective study was conducted of the 8 patients who had been treated at Department of Orthopaedics, The Third Affiliated Hospital to Peking University for Cierny-Mader type Ⅳ osteomyelitis of the lower extremity from June 2017 to December 2019. They were 6 males and 2 females, aged from 27 to 79 years (average, 54.6 years). The defects involved femoral shaft in 5 cases, femoral metaphysis in one, tibia shaft in one, and tibial metaphysis in one. The defect lengths ranged from 7.7 to 15.5 cm, averaging 10.2 cm. Stage one was local infection control and temporary stability reconstruction using Masquelet technique, stage two design and 3D printing of the prosthesis and stable pattern design, and stage three prosthesis implantation and rehabilitation. The ranges of motion of the knee and ankle were recoded postoperatively and the functions evaluated using the Johner-Wruhs scores.Results:The average follow-up time for the 8 patients was 12.6 months (from 6 to 18 months). The total treatment time from the first admission to the last discharge ranged from 62 to 125 days (average, 91.0 days), the time for stage one from 13 to 57 days (average, 28.7 days), that for stage two from 30 to 87 days(average, 48.3 d), and that for stage three from 28 to 84 days (average, 63.0 days). The infection was controlled and there was no recurrence, implant loosening or breakage. Seven patients were capable of full weight-bearing at 14.7 days (from 4 to 42 days) after surgery. One patient recovered full weight-bearing 6 months after surgery due to severe osteoporosis. Fine functional recovery was achieved in the 8 patients, with a range of motion from 0° to 100° for the knee and a range from 35° dorsal flexion to 40° toe flexion for the ankle. The Johner-Wruhs scores at the last follow-up showed 2 excellent, 5 good and one moderate cases.Conclusion:In repair of Cierny-Mader type Ⅳ long bone osteomyelitis, Masquelet technique and 3D printing can shorten the treatment process and allow for early recovery.

ObjectiveTo investigate the role of Notch1 gene in xenografted human cutaneous squamous cell (SCL-1) carcinoma. MethodsFifteen nude mice were divided into three groups, including untreated group(inoculated with SCL-1 cells treated with phosphate buffered saline), empty vector group (inoculated with SCL-1 cells transfected with empty vector) and Notch1 group(inoculated with SCL-1 cells transfected with Notch1 expression vector). All the mice were inoculated with SCL-1 cells(1 x 108/ml) of0.2 ml. Then, the growth of xenografted tumor was observed every other day. Fifteen days later, the mice were sacrificed, tumor tissue was dissected and subjected to terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay for the detection of cell apoptosis, reverse-transcription(RT)-PCR and Western blot for the examination of mRNA and protein expressions of Notch1, bcl-2 and bax, respectively. ResultsThe proliferation of xenografted tumor in Notch1 group was obviously inhibited compared with the untreated group. The weight of xenografted tumor in Notch1 group was significantly lower than that in the untreated group and empty vector group (0.574 ± 0.219 g vs. 2.642 ± 0.404 g and 2.606 ± 0.512 g, F= 26.642, P＜ 0.01). TUNEL assay demonstrated that the number of apoptotic cells per 500 cells in tumor tissue specimens was(87 ± 9) in Notch1 group, evidently higher than that in the untreated group(8 ± 2) and empty vector group(10 ± 3) (F = 194.266, P ＜ 0.05 ). Further, RT-PCR and Western blot revealed that the mRNA and protein expressions of Notch1 and bax were significantly upregulated, but those of bcl-2 were markedly downregulated in the Notch 1 group, with significant difference among the three groups(all P ＜ 0.05). ConclusionsNotch 1 gene can inhibit the growth of xenogra ffted human cutaneous squamous cell(SCL-1) carcinoma and induce SCL-1 cell apoptosis likely by upregulating bax expression and downregulating bcl-2 expression.

Objective To investigate the minimal invasive management of cystitis glandularis with invasion of the ureteric meatus.Methods The clinical data of 18 cases were reviewed.Among the 18 cystitis glandularis patients,12 cases were invasion of bilateral ureteric meatus and 6 of unilateral ureteric meatus.Operation or drug treatment was performed on the diseases that can cause cystitis glandularis such as bladder stone,bladder neck stegnosis,external urethral meatus stegnosis and benign prostate hyperplasia.Sensitive antibiotic was administrated in all cases.After placing ureter catheter,transurethral plasma electro-resection was carried out in five patients whose ureteric meatus could be identified.In addition,of thirteen patients with ureteral orifice unable to be identified,there were ten cases with normal renal function,mitomycin was injected under affected membrana mucosa,and then the patient with ureteral orifice identified underwent transurethral plasma electro-resection after placing ureter catheter.On the other hand,the patient whose ureteral orifice still could not be recognized undertaken transurethral electro-resection at first,during which the ureter catheter was put once ureteral orifice had been detected,otherwise,the ureter catheter should be placed through cystoscope if nephritic colic emerged and hydronephrosis aggravated after operation.In those patients with kidney dysfunction,the ureter catheter was put by ureter discission or ureter replantation at first,the transurethral electro-resection could not be executed until the renal function recovered.Following all these procedure above,bladder instillation of drugs regularly,anti-infection and symptomatic treatment were administrated.Results One patient combined with bladder adenocarcinoma received cystectomy,of the other patients,six cases recurred and underwent electrotomy again resulting in no relapse.All nephrohydrops vanished or relieved obviously,nevertheless,urinary tract infection,haematuria and bladder-ureter backstreaming as the cardinal complication developed in some cases.Conclusion In the management of cystitis glandularis encroaching ureteric meatus,total or partial cystectomy can be avoided if ureter draining freely can be ensured,motivation removal,antiinfection,injection of drug under mucosa and preoperative diuresis conduce to the achievement of ureter catheter placing,transurethral plasma electro-resection is still effective methods in treating these cystitis glandularis.

BACKGROUND: Recently biodegradable hydrogel has been extensively used to delivery anticancer drug and bioactive macromolecule. However, to protect the activity of the bioactive macromolecule, we need to obtain series of hydrogel which have milder hydrogelation conditions and shorter hydroglation time.OBJECTIVE: To prepare enantiomeric poly(L-Lactic acid) (PLLA)-poly(ethylene glycol (PEG)-PLLA/ poly(D-Lactic acid) (PDLA)-PEG-PDLA stereocomplex hydrogel which has shorter hydroglation time, to physically encapsulate a model drug-lysozyme and sustained release it from the hydrogel. METHODS: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were synthesized by ring-opening polymerization of L(D)-lactide using PEG as the initiator and Sn(Oct)2 as the catalyst. The triblock copolymers were characterized by 1H nuclear magnetic resonance, FT-IR and X-Ray diffractometry. A hydrogel was prepared from an aqueous mixture of PLLA20-PEG227-PLLA20 and PDLA21-PEG227-PDLA21 (10 wt% concentration) at room temperature for 12 hours. X-Ray diffractometry test was used to research the gelation mechanism. The release profile of the lysozyme as a model drug from the hydrogel was tested. The morphology of the freeze-dried hydrogel was investigated by scanning electron microscope. The cytotoxicity of the hydrogel was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay.RESULTS AND CONCLUSION: Triblock copolymers of PLLA-PEG-PLLA and PDLA-PEG-PDLA were obtained. Both the PEG and PLA blocks in the copolymers could crystallize, but the crystallization of the PEG block was predominant. The stereocomplex formation between the PLLA and PDLA blocks within the hydrogel was confirmed by the X-Ray diffractometry analysis. The release profile of the lysozyme from the hydrogel exhibited a sustained-release pattern with a duration period of 7 days. The hydrogel exhibited a 3D interconnected porous structure with 50-100 μm pore size after being freeze-dried. The mouse fibroblast cell viability percentage was 99.3% after the cells contacted with the 100% extracted liquid for 72 hours.