Effect of GTPBP4 silencing on radiosensitivity of EC9706 cells / 中华放射肿瘤学杂志

Objective:To investigate the effect of GTPBP4 silencing by RNA interference on the radiosensitivity of esphageal cancer EC9706 cells line.Methods:The expression data of GTPBP4 in esophageal cancer tissues was obtained from public Gene Expression Omnibus (GEO) database. Recombinant plasmid-mediated RNA interference (RNAi) was employed to transfect the esophageal cancer EC9706 cell to evaluate the influence of GTPBP4 silencing on the proliferation, apoptosis and radiosensitivity of esphageal cancer EC9706 cells. The expression levels of GTPBP4 mRNA and protein and apoptosis-associated proteins of Bax, cleaved caspase-9, cleaved caspase-3 and Bcl-2 were determined by qRT-PCR and Western blot. The cell proliferation was determined by MTT assay. The changes in cell apoptosis were detected AnnexinⅤ-FITC/PI double staining flow cytometry. The variations in radiosensitivity after radiation exposure were assessed by clone formation assay.Results:The expression level of GTPBP4 in the esophageal cancer tissues was significantly higher than that in the normal adjacent esophageal tissues ( P<0.001). qRT-PCR and Western blot demonstrated that the expression levels of GTPBP4 mRNA and protein in the GTPBP4-siRNA group were significantly lower than those in the blank and negative control groups (both P<0.001), suggesting that the plasmid was successfully transfected into the EC9706 cells. MTT assay indicated that the EC9706 cell proliferation rate was significantly inhibited ( P<0.001). Flow cytometry found that the apoptosis rate was significantly increased in the GTPBP4-siRNA group ( P<0.001). After GTPBP4 gene interference combined with radiotherapy, the cell sensitivity enhancement ratio was 1.716. The apoptosis rate of EC9706 cells was significantly increased in the GTPBP4-siRNA group ( P<0.001). The expression levels of apoptosis-associated proteins including cleaved caspase-9, cleaved caspase-3 and Bax were significantly up-regulated, whereas that of Bcl-2 was significantly down-regulated in the EC9706 cells in the GTPBP4-siRNA group ( P<0.001, P=0.001, P=0.001 and P=0.005). Conclusions:GTPBP4 gene is highly expressed in human esophageal cancer tissues. RNAi technology can effectively inhibit the expression of GTPBP4 gene in the EC9706 cells, thereby suppressing cell proliferation, inducing cell apoptosis and enhancing the radiosensitivity of cells.

AMG-102 inhibits proliferation and induces apoptosis of laryngeal squamous cell carcinoma cells by regulating c-Met/PI3K/Akt pathway / 中华肿瘤杂志

Objective@#To investigate the effects of c-Met inhibitor AMG-102 on the proliferation and apoptosis of laryngeal squamous carcinoma Hep-2 cells and the underlying mechanism.@*Methods@#Laryngeal squamous carcinoma cell line Hep-2 cells were treated with 2.5, 5 and 10 μmol/L AMG-102, respectively. The proliferation activities of Hep-2 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT). The apoptotic rate of Hep-2 cells was detected by flow cytometry analysis and Hoechst staining. The mRNA expression levels of apoptosis-related genes were detected by real-time quantitative polymerase Chain reaction (RT-qPCR), and the protein expressions of c-Met/PI3K/AKT pathway were detected by western blot.@*Results@#Compared with the control group, the proliferation rates of Hep-2 cells treated with 2.5, 5 and 10 μmol/L AMG-102 for 24 hours were (89.8±1.1)%, (79.8±1.0)% and (69.1±1.2)%, respectively; for 48 hours were (76.8±2.0)%, (60.2±1.1)% and (49.8±1.2)%, respectively; for 72 hours were (50.1±2.0)%, (41.5±1.1)% and (33.6±1.0), respectively, with significant differences (all P<0.05). The apoptotic rates of Hep-2 cells treated with 2.5, 5 and 10 μmol/L AMG-102 for 48 hours were (16.09±1.53)%, (27.51±2.02)% and (36.57±1.42)%, respectively, which were significantly higher than (3.62±0.10) % in the control group (all P<0.05). After treated with 2.5, 5 and 10 μmol/L AMG-102 for 48 hours, the relative expression levels of Bcl-2 mRNA in Hep-2 cells were 0.58±0.13, 0.38±0.12 and 0.20±0.13, respectively; the relative protein expression of p-Met were 80.0±3.8, 50.6±4.2 and 28.5±1.3, respectively; the relative protein expression of p-PI3K were 87.1±0.9, 54.2±1.2 and 21.0±1.2, respectively; the relative protein expression of p-AKT were 98.7±5.6, 56.9±3.2 and 32.2±4.3, respectively; which were significantly lower than those in the control group (all P<0.05). The relative expression levels of Bax mRNA were 1.78±0.13, 2.37±0.14 and 3.05±0.13, respectively, and the relative expression levels of caspase-3 mRNA were 1.98±0.14, 2.47±0.14 and 3.15±0.13, respectively, which were significantly higher than those in the control group (all P<0.05).@*Conclusion@#c-Met inhibitor AMG-102 could inhibit the proliferation and induce apoptosis of laryngeal squamous carcinoma Hep-2 cells by regulating the c-Met/PI3K/Akt pathway.

Exploring Anti-inflammation Mechanism of Pien Tze Huang via Regulating of Microglia Polarization / 中国实验方剂学杂志

Objective::To investigate the anti-inflammation mechanism of Pien Tze Huang (PTH) via regulating microglia polarization. Method::The experiment was divided into five groups, Blank, M1[lipopolysaccharide (LPS) 100 μg·L-1+ interferon-γ(IFN-γ) 10 μg·L-1], M1-PTH group[LPS 100 μg·L-1+ IFN-γ 10 μg·L-1+ PTH 0.4 g·kg-1], M2 group[interleukin-4 (IL-4) 20 μg·L-1], and M2-PTH group[IL-4 20 μg·L-1+ PTH 0.4 g·kg-1]. The concentration of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), and transforming growth factor-β1(TGF-β1) in the culture supernatant were measured by enzyme-linked immunosorbent assay (ELISA), the levels of inducible nitric oxide synthase(iNOS) and arginine-1 (Arg-1) mRNA were detected by real-time fluorescent quantitative polymerase chain reaction technique(Real-time PCR), and the expression levels of p-STAT1, p-STAT3, iNOS, p-STAT6, and Arg-1 were detected by Western blot. Result::The concentration of NO and TNF-α of the culture supernatant, the level of iNOS mRNA, as well as the level of p-STAT1, p-STAT3 and iNOS in M1 group, which were significantly increased(P<0.01) .Compared with blank group, but the concentration of NO and TNF-α were down-regulated(P<0.01), and iNOS mRNA(P<0.05), as well as the expression of iNOS, p-STAT1, and p-STAT3 was decreased (P<0.05, P<0.01) after the invention of PTH in M1-PTH group compared with M1 group. The concentration of IL-10 and TGF-β1 in the culture supernatant, the mRNA level of Arg-1, as well as the levels of p-STAT6 and Arg-1 were significantly increased in M2 group when compared with Blank group, addition to the concentration of IL-10 and TGF-β1 were up-regulated(P<0.05, P<0.01), and the expression of Arg-1 mRNA, the level of Arg-1, p-STAT6 were enhanced(P<0.05, P<0.01) in M2-PTH group compared with M2 group. Conclusion::PTH plays an anti-inflammatory role via regulating microglia polarization.

Effect of c-Met inhibitor AMG-102 on radiosensitivity in laryngeal squamous carcinoma cells / 中华肿瘤杂志

Objective@#To investigate the effect of c-Met inhibitor AMG-102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells.@*Methods@#The effects of AMG-102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep-2 and KBV200 were detected by 3-(4, 5-dimethy-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay and colony formation assay, respectively. The apoptosis of Hep-2 and KBV200 cells was detected by flow cytometry. The expression levels of c-Met, phospho-Met (p-Met), cleaved caspase-3 and Akt/p-Akt, Erk/p-Erk were detected by Western blot. Specific small interfering RNA targeting c-Met or plasmid of c-Met were transfected into Hep-2 and KBV200 cells to investigate the cell sensitivity to AMG-102.@*Results@#Compared with KBV200 cells, Hep-2 cells were more sensitive to AMG-102 with IC50 of 14 and 9 μmol/L, respectively. The relative expression levels of c-Met and p-Met proteins in Hep-2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells (171.24±1.00 and 115.37±0.56, respectively, P<0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p-Met protein in KBV200 cells. The results showed that AMG-102 significantly reduced the expression of p-Met in KBV200 cells treated with HGF (P<0.001). Compared with the dimethyl sulfoxide (DMSO) group, AMG-102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep-2 cells (SER=1.28, P<0.001). However, AMG-102 had little effect on the radiosensitivity of KBV200 cells (SER=1.18, P=0.002). Compared with the 4 Gy radiotherapy alone group and the 5 μmol/L of AMG-102 alone treatment group, the apoptosis rate of Hep-2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase-3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase-3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p-Met, p-Akt and p-Erk were significantly decreased in the 4 Gy radiotherapy group, 5 μmol/L of AMG-102 treatment group and combined treatment group of Hep-2 cells. And the levels of p-Met, p-Akt and p-Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 μmol/L of AMG-102 treatment alone group. By contrast, in KBV200 cells, the expression of p-Met, p-Akt and p-Erk in each group was not changed. The relative expression of p-Met in Hep-2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p-Met was slightly increased after radiotherapy at 30 min and 1 h (P<0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P<0.05 for all). By contrast, the expression of p-Met in KBV200 cells did not change with time after radiotherapy (P>0.05). The sensitivity of Hep-2 cells to AMG-102 was decreased after silencing of c-Met, while the sensitivity of KBV200 cells to AMG-102 was not significantly changed (P>0.05). Moreover, the radiosensitivity of Hep-2 cells in c-Met knockdown group had a slightly increasing trend (SER=1.07, P=0.068). After the treatment with 10 μmol/L of AMG-102, the proliferation rate of c-Met ectopically expressed KBV200 cells was 60.05%±3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P<0.001). The results suggested that the overexpression of c-Met in KBV200 cells increased the radiosensitivity to AMG-102, whereas depletion of c-Met resulted in resistance to AMG-102 in Hep-2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c-Met showed a decreased trend (SER=0.7, P=0.005).@*Conclusions@#c-Met inhibitor AMG-102 has a significant inhibitory effect on the proliferation of c-Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c-Met receptor is more effective in c-Met overexpressed subtype of laryngeal squamous cell carcinoma.

Insights of Chinese medicine on ventricular remodeling: Multiple-targets, individualized-treatment / 中国结合医学杂志

Ventricular remodeling (VR) can be induced by myocardial injury, leading to progressive cardiac dysfunction and heart failure, and is associated with high morbidity and mortality. Despite being studied for more than 3 decades, current therapeutic strategies still remain unsatisfactory in effificacy, expensive, and with side effects and drug resistances. Chinese medicine (CM) has been used to treat heart diseases for thousands of years. This article reviews the published studies on the mechanisms and therapeutic applications of CM in VR. The major aspects include: mechanistic studies of VR, molecular biology and myocardial functional studies of CM therapies on VR, and mechanism of CM therapies on VR.

Case-control study on the iliac bone flap transplantation with deep circumflex iliac artery and quadratus femoris bone flap transplantation for the treatment of Garden III/IV femoral neck fracture of young and middle-aged patients / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effects between hip anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery and posterior K-L approach combined with quadratus femoris bone flap transplantation for the treatment of femoral neck fracture of Garden III-IV in young and middle-aged patients.</p><p><b>METHODS</b>From January 2004 to January 2011,46 patients with femoral neck fractures were treated by two kinds of operation. Among them, 20 cases were treated with anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery, included 12 males and 8 females with an average age of (32.1 ± 7.3) years old, involved 12 cases of Garden III and 8 cases of Garden IV. The other 26 cases were treated with posterior K-L approach combined with quadratus femoris bone flap transplantation, included 20 males and 6 females with an average age of (37.8 ± 6.9) years old, involved 16 cases of Garden III and 10 cases of Garden IV. The index of hospitalization (hospitalization time, total cost, operative time, intraoperative blood loss, postoperative complications), the quality index of operation (fracture reduction, position of internal fixation, fracture healing time, nonunion and femoral head necrosis) of two groups were observed and compared. Hip joint function were evaluated by Harris score.</p><p><b>RESULTS</b>All patients were followed up from 28 to 41 months with an average of 36 months. The intraoperative blood loss of group S-P (92.3 ± 10.4) ml was less than that of group K-L (132.4 ± 11.2) ml, there was significant difference between two groups (P < 0.05). The operation time of group S-P (81.4 ± 9.2) min was more than that of group K-L (67.1 ± 4.5) min, the difference was statistically significant (P < 0.05). One case in group S-P and 9 cases in group K-L appeared postoperative complications, there was significant difference between two groups (P < 0.05). The fracture healing time of S-P group (83.5 ± 7.3) d was shorter than that of group K-L (103.2 ± 12.6) d, there was significant difference between two groups (P < 0.05). At 30 months after operation, there were significant difference in Harris scoring between two groups (P < 0.05).</p><p><b>CONCLUSION</b>Anterior S-P approach combined with iliac bone flap transplantation with deep circumflex iliac artery for treatment of femoral neck fracture of Garden III-IV of young and middle-aged patients, it has characteristics in clear anatomic and easy to operate. As compared with K-L approach, S-P approach can better reserve residual blood supply of femoral neck. While combining with the iliac bone flap transplantation with deep circumflex iliac artery, it could better reconstruct the blood supply of femoral neck to promote fracture healing.</p>

Repair of scars in submaxillary region using expanded forehead axial flaps with fascia pedicles carrying bilateral frontal branches of superficial temporal artery and vein / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the feasibility of applying expanded forehead axial flaps with fascia pedicles carrying bilateral frontal branches of superficial temporal artery and vein (expanded forehead axial flap with double pedicles in brief, EFAF-DP) in repairing scars in submaxillary region.</p><p><b>METHODS</b>Sixteen patients with mandibular scars hospitalized in Department of Burns and Plastic Surgery of the First Hospital Affiliated to Fuzhou General Hospital in Nanjing Military Area Command from July 2005 to December 2009 were repaired with EFAF-DP. The operation consisted of 3 stages. Before operation, the location and course of superficial temporal arteries and veins (STAV) and their frontal and parietal branches were identified with Ultrasonic Doppler blood flow detector. In stage I, STAV were dissected from the frontalis muscle as a pedicle to form a skin soft tissue space to hold the dilator of a proper size. In stage II, after gradual dilation by repeated filling with saline, the dilator was removed. EFAF-DP was dissected to repair mandibular scar. Donor site was closed with sutures. In stage III, flap pedicles were divided and pruned.</p><p><b>RESULTS</b>Flap sizes ranged from 25 cm × 6 cm to 33 cm × 16 cm. The duration of dilation was 3-5 months, with 3.6 months in average. Ten patients underwent the operation of EFAF-DP transplantation and cervical skin dilatation. All flaps survived with healing of wounds. Disorder of venous return at the distal end of one flap was seen after second stage surgery, and it was corrected after comprehensive treatment including relieving spasm and improving venous return. Donor site wounds healed with normally grown hair without cicatricial alopecia along the hairline. Few hairs grew around mandible in one female patient out of the three (no hair grew on flaps of other two patients). This female patient and two male patients requesting for beard plasty received laser depilation treatment 1 to 3 months after discharge, with good result. Other male patients received no special treatment for their beard, and they shaped their beard with shaver. Sixteen patients were followed up for 6 to 24 months, and the shape of the flaps and beard (excluding female patients) were satisfactory with good appearance, satisfactory skin color and texture. The mobility of neck was obviously improved.</p><p><b>CONCLUSIONS</b>EFAF-DP provides bigger areas of a thin flap besides promoting vascularization of new vessels of flap. Extra expanded skin can be directly sutured at the fringe of hairline, which makes skin grafting unnecessary, and decreases the incidence of secondary deformity in donor sites. Some hair carried by the flaps can be directly used for beard reconstruction after rotation to help the male patients have a better appearance.</p>

The treatment of cicatricial alopecia after burn with the technique of synchronously perforating and transplanting hair follicular-units / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the effect of the technique of synchronously perforating and transplanting hair follicular-units in the treatment of cicatricial alopecia after burn.</p><p><b>METHODS</b>One hundred and sixty-six patients with 217 bald scar areas after burn were treated with above-mentioned technique from January 2002 to April 2008. Scalp strips, with conforming the necessity for grafting, were harvested from the occipital or temporal region. A series of follicular-units, each composing 1 - 3 hairs, were dissected from the strips under microscope or magnifying glass. Size-matching micro-slots were made in the scarred recipient area with 16 - 20 G needles to accept the grafts. The prepared follicular-unit was synchronously implanted into the bottom of the micro-slot as the needle being withdrawn. Patients who were not satisfactory with the density of hairs after I stage surgery underwent II stage surgery a half year later. Ten recipient areas with clear boundary in 10 patients were optionally chosen to observe the density of follicular-units and hair amount with naked eyes after I stage surgery. Survived transplanted hairs in above-mentioned 10 areas were counted to calculate hair survival rate at follow-up. Patients' postoperative satisfaction ratings were surveyed with questionnaire.</p><p><b>RESULTS</b>In one half of the patients, treatment was finished after I stage surgery, the other one half received 2 stages of surgery. The follicular-unit density reached 15 - 25 grafts/cm(2) with 40 - 70 hairs/cm(2) after I stage surgery. All patients were followed up for over 8 months. Grafted hairs grew well in a natural way. 96.5% mean hair survival rate was observed in the 10 recipient areas. From patients who received only I stage surgery, 61 patients (73.5%) were very satisfactory and 22 patients (26.5%) satisfactory with the results. From the other half of patients, 76 patients (91.6%) were very satisfactory and 7 patients (8.4%) satisfactory with the results.</p><p><b>CONCLUSIONS</b>The technique of perforating and transplanting follicular-unit hair synchronously is safe and effective with less surgery-induced injury and less bleeding. Hairs transplanted on cicatricial alopecia area with this technique grow well with high survival rate.</p>

L-Proline as an efficient and reusable promoter for the synthesis of coumarins in ionic liquid / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The effect of L-proline as a promoter on the condensation reaction of salicylaldehyde or its derivatives with ethyl acetoacetate in neutral ionic liquid [emim]BF4 was studied. All reactions were carried out under mild reaction conditions and achieved high yields. Moreover, the ionic liquid containing L-proline could be recycled and reused for several times without noticeably decreasing in productivity. The results show that the L-proline-[emim]BF4 system has a potential in contribution to the development of environmentally friendly and inexpensive processes in organic syntheses.

Clinical application of the scapular free flap extended to the upper arm / 中华整形外科杂志

<p><b>OBJECTIVE</b>To apply the scapular free flap extended to the upper arm for resurfacing the face and neck, as well as the upper lip in one stage.</p><p><b>METHODS</b>The scapular free flap was designed with extended portion to the posterior and interior part of the upper arm. Then the free flap was transferred to resurface the face and neck with the routine portion and to resurface the upper lip with the extended portion.</p><p><b>RESULTS</b>6 cases with extensive upper lip, facial and cervical burn scar were treated with the extended scapular free flaps. The flap size ranged from 22 cm x 11 cm to 40 cm x 9.5 cm (36.57 cm x 10.20 cm in average) for the routine portion and from 7 cm x 4 cm to 12 cm x 4 cm (10.32 cm x 3.67 cm in average) for the extended portion. All flaps survived completely.</p><p><b>CONCLUSIONS</b>There are direct communicating branches ("choke vessel") between the circumflex scapular artery (CSA) and the posterior humeral circumflex artery (PHCA). When the blood supply of PHCA is cut off, the CSA can provide blood supply through the communicating branches to the upper arm skin area previously nourished by PHCA. So the blood supply of the extended portion of the scapular free flap is not only from the branches of CSA, but also from the direct communicating branches between the CSA and PHCA. The extended scapular free flap has a reliable blood supply and can be applied to construct the facial and cervical scar contraction with the extended portion to resurface the upper lip. The satisfactory result can be expected.</p>

A "silicone suture" technique aided to scalp reduction for the treatment of scarring alopecia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate a "silicone suture" technique for enhancing the effect of scalp reduction.</p><p><b>METHODS</b>Under the local anesthesia, when an incision was made in the midline of the lesion, the dissection was carried out underlying the sub-galea on both sides of the lesion, as far as the width of the lesion. A 3 mm silicone suture in diameter was placed in the galea beyond the lesion. After the first suture bite was anchored in the tissue at one end, the suture device was continued across the midline in such a way as a running, buried, horizontal mattress suture and it was brought out to the skin surface through the deep tissue at the another end of the lesion with a locker. The extra-tissue of the lesion was then excised and the wound was directly closed in layers. After one week of the operation, the silicone suture was gradually tightened in 2-3 times a week for about 3-5 weeks, until both sides of the lesion were approximately closed. The device was there after removed and the wound was directly closed in layers after the scar was excised.</p><p><b>RESULTS</b>Between October of 1999 and March of 2006, 12 scarring-scalp patients, 7 males and 5 females, were treated by using the silicone suture device without complications. The excised defects were 5-10.5 cm in width. The stretching period was 26.4 days in mean. With the following-ups over 3 months, no hypertrophic scar and widening scar cases appeared.</p><p><b>CONCLUSIONS</b>The silicone suture as an alternative device for tissue extension could be a safe, simple, effective and economical device. It could significantly enhance the efficiency of scalp reduction.</p>

Repair of the facial defects using the expanded induced prefabricated skin flap of the retroauricular and mastoid process region based on the superficial temporal vascular bundle / 中华整形外科杂志

<p><b>OBJECTIVE</b>To provide an ideal method for flap prefabrication.</p><p><b>METHODS</b>The superficial temporal fascial flap has been elevated based on the superficial temporal vessels during the first-stage procedure. A subcutaneous tissue pocket with appropriate site was formed in the retroauricular and mastoid process region. The fascial flap was transferred into the pocket and fixed properly. The tissue expander was placed under the fascial flap. When the expanding process has been finished, the expander was removed and the expanded induced prefabricated skin flap of the retroauricular and mastoid process region pedicled on the superficial temporal vascular bundle was elevated and transferred to repair the facial skin defect.</p><p><b>RESULTS</b>There were nine cases in the group. Facial defects after resection of the melanotic nevus was repaired in 2 cases and facial defects after resection of the facial haemangioma and scar were repaired in 2 and 5 cases respectively. Pedicle length of the superficial temporal fascial flap was ranged from 5.5 cm to 7 cm (mean length 6.2 cm). The size of the fascial flaps was ranged from 4 cm x 3 cm to 7 cm x 7 cm (mean size 5.7 cm x 4.9 cm). The size of the prefabricated skin flaps was ranged from 5 cm x 5 cm to 8.0 cm x 7.5 cm (mean size 6.4 cm x 6.1 cm). The average time of the tissue expansion process is 16.1 weeks. All flaps survived postoperatively and the donor sites of the flaps were appropriated directly in 5 cases. The split-thickness skin grafting was used to recover the donor site defects in 4 cases.</p><p><b>CONCLUSIONS</b>The superficial temporal fascial flap owns the following advantages: the vascular pedicle is much longer and vascular supply is plentiful, and it is convenient to transfer. Meanwhile, the skin of the retroauricular and mastoid process region is most similar to that of the face in texture, color and depth. For the patients selected strictly, the technique mentioned above is somewhat an ideal method for facial defect repair.</p>

The penis prolongation and augmentation combined with autologous granular fat injection and silicone implantation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To prevent the retraction of the penis after prolongation and augmentation.</p><p><b>METHODS</b>After all the superficial and part of the deep suspensory ligament amputation, we implanted the silicon sheet (the length 2.3-3.6 cm, the width 1.5-2.5 cm, the thickness 2-3 mm) and injected autologous granular fat (30-48 ml) into penis.</p><p><b>RESULTS</b>16 patients (age 22-63 years, averagely 38 years) underwent this kind operation, the prolongation length is 1.8-5.1 cm, the average was 2.91 cm, the increased diameter of penis was 0.6-1 cm, the average is 0.85 cm, the following period is 3 months to 2 years. The results are satisfactory with the penis retraction less than 8%, and less than 10% decrease in diameter.</p><p><b>CONCLUSIONS</b>This method is an ideal way of the penis prolongation and augmentation, the implantation of the silicon sheet is effective way to prevent the retraction of the penis.</p>

A clinical study on diagnosis and treatment of renal tuberculosis:a report of 82 cases / 中国基层医药

Objective To investigate the clinical characteristics on diagnosis and treatment of renal tubercu- losis.Methods Clinical data of 82 patients with renal tuberculosis were retrospectively reviewed.Results All of the 18 cases who received medication recovered completely.64 cases undergoing surgery were pathologically diagnosed to have renal tuberculosis,of which 2 cases developed ureteral stump syndrome.Conclusion Urine PCR-TB-DNA re- mained the primary diagnostic method before operation.Computed tomography(CT)and magnetic resonance urogra- phy(MRU)could be used in diagnosis of renal tuberculosis.When the non-functioning kidney was resected,per- inephrit fat and the involved ureter should be concomitantly resected as much as possible.

Proteomic analysis of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the mechanism of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells.</p><p><b>METHODS</b>In this study, the proteins extracted from paclitaxel-induced apoptotic MCF-7 cells were analyzed by 2-dimentional gel electrophoresis (2-DE), and compared with those from untreated MCF-7 cells. The differential proteins were identified by mass spectrometry.</p><p><b>RESULTS</b>At 24 hour after paclitaxel (100 nmol/L) treatment, MCF-7 cells were collected and extracted the whole proteins. Seventeen up-regulated or down-regulated proteins were found by analysis of the differential proteomic 2-DE map. Six of them were identified by mass spectrometry. They were enolase 1, chloride intracellular channel 1, keratin 8, ribosomal protein S12, galectin-1 and histidine triad nucleotide binding protein, respectively.</p><p><b>CONCLUSION</b>We effectively found the changed proteins in the process of paclitaxel-induced apoptosis in MCF-7 human breast carcinoma cells by proteomic techniques. These up-regulated or down-regulated proteins are important molecules for our further research about the mechanism of paclitaxel-induced apoptosis.</p>

Binding interactions of pefloxacin mesylate with bovine lactoferrin and human serum albumin / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Förster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy.

An experimental study on the culture of human oral mucosa epithelium using autologous serum / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the biological properties of cultured human oral mucosa epithelium using autologous serum in order to provide a new material for tissue engineering urethra.</p><p><b>METHODS</b>The cultured oral mucosa epithelium was respectively transplanted beneath the skin, above the deep fascia and in the wound of the athymic mice. The specimens were taken at 2, 3, 4, and 6 weeks posttransplantation, and processed for (1) immunofluorescence anti-HLA staining to determine graft acceptance, and (2) anti-human IV collagen and antihuman laminin immunohistochemical staining procedures to indicate the basement membrane formation.</p><p><b>RESULTS</b>All the grafts survived and grew very well. The grafts were positive to anti-HLA. Collagen type IV and laminin were detected at the dermo-epidermal junction in all groups from day 14, and increasing in density up to day 21.</p><p><b>CONCLUSIONS</b>he cultured human oral mucosa epithelium by autologous serum could develop an excellent functional epithelium tissue, which would be used to reconstruct urethra and repair wound.</p>

An animal model of venous congested flap for long-term microcirculation study / 中华整形外科杂志

<p><b>OBJECTIVE</b>To set up a venous congested flap model to study the mechanism of necrosis through long-term microcirculation observation.</p><p><b>METHODS</b>A specially deviced chamber was assembled to one side of the ears in an adult white rabbit, about 7 approximately 10 days after the operation the congested flap model was made and the microcirculatory status of the flap was dynamically observed under a vivo-microscope for a long time.</p><p><b>RESULTS</b>The venous crisis phenomenon of flap was well studied and the microcirculation of the flap was observed carefully, finally the variational rule of the congestion flap microcirculation was made clear.</p><p><b>CONCLUSIONS</b>The model could well simulate the venous crisis flap in clinic, and the microcirculation could also be observed for a long time.</p>

Construction of phage display antibody library to MCF-7 cells and screening of single-chain antibodies against breast cancer cells / 生物工程学报

The aim of this study is to construct a phage display single-chain variable fragment (scFv) library against breast cancer cells and screen the specific antibodies against MCF-7 cells from the library. The BALB/C mice were immunized with MCF-7 cells. Total RNA of spleens was isolated. The heavy-chain (VH) and light-chain variable region genes (VL) of the antibodies were amplified by RT-PCR and joined into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The assembled scFv fragments were cloned into the phagemids(pCANTAB5E) and the recombinant phagemids were used to transform competent E. coli TG1. The transformed TG1 cells were infected by helper phage M13KO7 and the recombinant phagemids were rescued. The scFv fusion proteins were displayed on the surfaces of the recombinant phages. A phage display antibody library of repertoire of 1.2 x 10(6) clones was constructed. The specific antibodies against MCF-7 cells were enriched by 75 times after five rounds of affinity selection. Ten recombinant phages clones that exhibited specific binding to MCF-7 cells were identified. The specificity of those phage clones was analyzed by reactivity against HepG2 cells and Hela cells by ELISA. One of the selected phage clones against MCF-7cells was used to infect E. coli TOP10 to produce the soluble scFv antibodies after induction with IPTG. The strategy of construction and screening of antibody library directed against the whole tumor cells described in this report should be generally applicable to generate tumor cell-specific antibodies.

Experimental study on inhibition of skin graft contraction with papaverine cream / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of exteral use of papaverine cream on enhancement of skin graft.</p><p><b>METHODS</b>Five mini-pigs were used for the study. 20 full-thickness wounds were created on the back of each animal. Along the midline of the abdomen, a 40 cm x 2 cm full-thickness skin graft was harvested and the wound sutured.The full-thickness graft was prepared and trimmed to 2 cm x 2 cm of 0.6 mm thickness split-skin grafts, which were transplanted to each wound with tie-over bolsters. The sutures were removed 2 weeks after the operation. Twelve pairs of 100%-survived skin grafts were selected and divided into group A (the left side) and group B (the right side), with each pair locating on the same and opposite position. From the day of suture removal, 2% papaverine cream was used to group A while plain cream was used to group B. The grafts were measured and observed for healing condition at the time of suture removal and the first, second, third, fourth, fifth, and sixth month. The surface area of the graft was measured and the shrinking ratio was calculated. After the animals were killed at the sixth month, samples of the skin grafts were taken and processed with 10% formalin, routine paraffin wax and Hematoxylin-eosin staining. The tissue structure was observed and the results were analyzed statistically.</p><p><b>RESULTS</b>The grafts in two groups did not have significant differences at the time of suture removal. Observations from the first to the sixth month showed that the two groups had significant differences in skin graft contracture and histological changes. HE stains demonstrated that the skin grafts in group A had less fibroblasts, more microvessels and orderly-arranged collagenous fibers.</p><p><b>CONCLUSIONS</b>External use of papaverine cream can inhabit the contraction of skin grafts. It is a safe, effective, simple, and reliable method,which has the advantages of easy delivery,cheapness, less injury and infection,and no side effects.</p>