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Objective: To observe the analgesic effect of Tuina by pressing and kneading the Huantiao (GB30) acupoint on rats with chronic constriction injury (CCI) and to explore the analgesic mechanism of Tuina on sciatica rats., Methods: Thirty-two SPF male SD rats weighing 180 to 220 g were randomly divided into fore groups:blank group (without any treatment), sham group (only exposed without sciatic nerve ligating), model group (sciatic nerve ligating) and Tuina group (manual intervention after lsciatic nerve ligating). The CCI model was prepared by ligating the right sciatic nerve of the rats, on the third day of modeling, the rats in the Tuina group were given pressing and kneading the Huantiao (GB30) point for 14 days, and the changes of paw withdrawal threshold(PWT), paw withdrawal latency(PWL) were measured before and on the 1st, 3rd, 7th, 10th, 14th and 17th days after modeling. The changes of sciatic functional index(SFI) were measured before and on the 1st and 17th day after modeling. The morphological changes of the sciatic nerve were observed by hematoxylin-eosin(HE) staining;and the differences in NF-κB protein expression in the right dorsal horn of the spinal cord of rats were detected., Results: Following modeling, there was no significant difference in PWT, PWL and SFI between the blank group and the sham group ( P >0.05), but the PWT, PWL and SFI of the model group and the Tuina group decreased significantly ( P <0.01). After manual intervention, the pain threshold of rats in Tuina group increased. On the 8th day of manual intervention (the 10th day after modeling), PWT in Tuina group increased significantly compared with that in model group ( P <0.01). On the 5th day of manual intervention (the 7th day after modeling), the PWL of the massage group was significantly higher than that of the model group ( P <0.01). The pain threshold of rats in Tuina group continued to rise with the continuous manipulation intervention. After 14 days of manipulative intervention, the sciatic nerve function index of rats in the Tuina group increased significantly( P <0.01). Compared with the blank group and sham group, the myelinated nerve fibers of sciatic nerve in the model group were disordered and the density of axons and myelin sheath was uneven. Compared with the model group, the nerve fibers of rats in the Tuina group were gradually continuous and the axons and myelin sheath were more uniform than those in the model group. Compared with the blank group and sham group, the expression of NF-κB protein in the right spinal dorsal horn of the model group was significantly increased( P <0.01). Compared with the model group, the expression of NF-κB protein in the right spinal dorsal horn of rats in Tuina group decreased significantly( P <0.01)., Conclusion: Pressing and kneading the Huantiao (GB30) point restores nerve fiber alignment;and improves the PWT、PWL and SFI in the CCI model by decreasing NF-κB p65 protein expression in the spinal dorsal horn. There fore, Tuina demmstrates an analgesic effect and improves the gait of rats with sciatica.

Galli Gigerii Endothelium Corneum(GGEC) is commonly used for the clinical treatment of indigestion, vomiting, diarrhea, and infantile malnutrition with accumulation. In recent decades, omnivorous domestic chickens, the original source of GGEC, has been replaced by broilers, which may lead to significant changes in the quality of the yielding GGEC. Through subjective and objective sensory evaluation, biological evaluation, and chemical analysis, this study compared the odor and quality between GGEC derived from domestic chickens and that from broilers. The odor intensity between them was compared by odor profile analysis and it was found that the fishy odor of GGEC derived from domestic chickens was significantly weaker than that of GGEC from broilers. Headspace-solid phase microextraction-gas chromatography-triple quadrupole tandem mass spectrometry(HS-SPME/GC-QQQ-MS/MS) suggested that the overall odor-causing chemicals were consistent with the fishy odor-causing chemicals. According to the odor activity va-lue and the orthogonal partial least squares discriminant analysis(OPLS-DA) result, dimethyl trisulfide, 2-methoxy-3-isobutylpyrazine, and 2-methylisoborneol were responsible for the fishy odor(OAV≥1) and the content of fishy odor-causing chemicals in GGEC derived from broilers was 1.12-2.13 folds that in GGEC from domestic chickens. The average pepsin potency in GGEC derived from broilers was 15.679 U·mg~(-1), and the corresponding figure for the medicinal from domestic chickens was 26.529 U·mg~(-1). The results of pre-column derivatization reverse-phase high-performance liquid chromatography(RP-HPLC) assay showed that the content of total amino acids and digestion-promoting amino acids in domestic chickens-derived GGEC was 1.12 times and 1.15 times that in GGEC from broilers, and the bitter amino acid content was 1.21 times folds that of the latter. In conclusion, GGEC derived from domestic chickens had weaker fishy odor, stronger enzyme activity, higher content of digestion-promoting amino acids, and stronger bitter taste than GGEC from broilers. This study lays a scientific basis for studying the quality variation of GGEC and provides a method for identifying high-quality GGEC. Therefore, it is of great significance for the development and cultivation of GGEC as both food and medicine and breeding of corresponding varieties.

Despite the distinctive characteristics and remarkable efficacy, animal medicine is stenchy, which decreases the comp-liance of patients. At the moment, the research on the method for deodorizing animal medicines lags behind. To be specific, the components related to the odor and the basic properties transformation of the components are unclear and there is a lack of specific deodorizing method. This study aims to clarify the main components related to the stench of animal medicine, such as aldehydes, amines, trimethylamines and sulfur compounds, and their basic properties, and to explore their metabolism and transformation in vivo and in vitro, which is expected to serve as a reference for the research on deodorization of animal medicine and development of new techniques.

Objective: To investigate the correlation between the changes of cervical curvature and atlantoaxial instability., Methods: The correlation between the changes of cervical curvature and atlantoaxial instability was retrospectively studied in 50 outpatients with abnormal cervical curvature (abnormal cervical curvature group) from January 2018 to December 2019. There were 24 males and 26 females in abnormal cervical curvature group, aged from 18 to 42 years old with an average of(30.62±5.83) years. And 53 patients with normal cervical curvature (normal cervical curvature group) during the same period were matched, including 23 males and 30 females, aged from 21 to 44 years with an average of(31.98±6.11) years. Cervical spine X-ray films of 103 patients were taken in lateral position and open mouth position. Cervical curvature and variance of bilateral lateral atlanto-dental space(VBLADS) were measured and recorded, Pearson correlation coefficient analysis was used to study the correlation between the changes of cervical curvature and atlantoaxial instability., Results: Atlantoaxial joint instability accounted for 39.6%(21/53) in normal cervical curvature group and 84.0%(42/50) in abnormal cervical curvature group. There was significant difference between two groups( P <0.01). VBLADS in abnormal cervical curvature group was (1.79±1.01) mm, which was significantly higher than that in normal cervical curvature group(0.55±0.75) mm( P <0.01). Pearson correlation coefficient analysis showed that the size of cervical curvature was negatively correlated with VBLADS., Conclusion: Cervical curvature straightening and inverse arch are the cause of atlantoaxial instability, the smaller the cervical curvature, the more serious the atlantoaxial instability.

Phyllanthi Fructus, a unique Chinese and Tibetan medicinal plant with both edible and medical values, has high potential of cultivation and development. The resources of Phyllanthi Fructus in China are rich, mainly distributed in Yunnan, Sichuan, Fujian, Guangdong, Guangxi, etc. Phyllanthi Fructus is widely used in the clinical practice of Chinese medicine and plays an important role in Tibetan medicine, Uyghur medicine, Yi medicine, and Mongolian medicine. Phyllanthi Fructus mainly contains phenolic acids,tannins, terpenes, sterols, fatty acids, flavonoids, amino acids and other compounds. Modern pharmacological studies show that Phyllanthi Fructus has antioxidant, anticancer, blood lipid-lowering, liver protective, antimicrobial, anti-inflammatory, and immune regulatory activities. In this paper, the research status of Phyllanthi Fructus was reviewed from the aspects of herbal textual research,chemical composition, and pharmacological action. The quality markers(Q-markers) of Phyllanthi Fructus were predicted and analyzed from the aspects of biogenic pathway, specificity and measurability of chemical components, efficacy, properties, new clinical uses, drug-food homology, and transformation of polyphenols. The results will provide a scientific basis for the quality control, quality evaluation, and standard formulation of Phyllanthi Fructus.

Indigo Naturalis has a long history of medicinal use with particularity and complexity in its processing. Before the Ming dynasty,Indigo Naturalis was extracted from the top layer of zymotic fluid,called &quot; purified Indigo Naturalis&quot;. In modern processing,the precipitate &quot; crude Indigo Naturalis&quot; is dried to produce Indigo Naturalis after impurity removal. The form of Indigo Naturalis slices has undergone significant changes in ancient and modern times. In view of this,the quality comparison between crude Indigo Naturalis and purified Indigo Naturalis was conducted in this study with modern analytical techniques. Firstly,chemical composition was analyzed with UPLC-Q-TOF-MS,and the chemical composition of scent with HS-SPME/GC-MS/MS. The content of indigo,indirubin,total ash,and water-soluble extract was determined as well as the inorganic composition in crude Indigo Naturalis and purified Indigo Naturalis. Then,their microscopic morphology was observed and the surface element composition was investigated. Finally,the antipyretic activities of crude Indigo Naturalis and purified Indigo Naturalis were compared in the fever rat model induced by lipopolysaccharide and 2,4-dinitrophenol. The results demonstrated that the purified Indigo Naturalis had a faster and more lasting antipyretic effect,while the crude Indigo Naturalis had almost no antipyretic effect. This study is of great significance to the research on processing technology of Indigo Naturalis and provides reference for the formulation of its quality standards,production specifications and calibration procedures.

Phyllanthus emblica is a kind of traditional medicine and medicinal and edible plant, with rich variety resources and high development value. It is a key poverty alleviation variety in China at present. As P. emblica processing industry is rising gradually in recent years, in order to fully develop and utilize its industrial resources, this paper systematically introduces current comprehensive development and utilization of P. emblica, discusses the problems in P. emblica processing industry, and puts forward comprehensive development and utilization strategies and industrial models in terms of cultivation, breeding, grading, quality evaluation and waste recycling, so as to provide a certain reference for promoting the high-quality development of P. emblica industry in China.

Objective: To investigate the impact of hypoxic-ischemic brain injury (HIBI) on brain development in neonatal rats of different sexes. Methods: From January 1 to December 31, 2018, 60 7-day-old SD rats were randomly divided into HIBI-F group (20 rats), HIBI-M group (20 rats), and control group (20 rats, 10 females and 10 males). The animal model of HIBI was established with Rice-Vannucci method, with the rats' left common carotid artery double-ligated and severed. The rats were then placed in an incubator and exposed to a hypoxic gas mixture (8% O(2), 92% N(2)) for 90 minutes. No intervention was given to the control group. Two weeks after HIBI, the motor development was evaluated by footprint analysis, the residual brain volume was measured by brain magnetic resonance imaging (MRI), and the damage of synaptic ultra structure was analyzed by transmission electron microscope. One-way ANOVA or χ(2) test was used for inter-group statistical analysis, and paired sample t test was used to compare the bilateral step length and toe distance of rats in the same group. Results: The mortality rate of HIBI-F was significantly higher than that of HIBI-M (20%(4/20) vs. 10%(2/20), χ(2)=40.000, P =0.001). The right step length and toe distance in HIBI-M group and HIBI-F group were significantly shorter than those in control group ((7.5±0.3) cm and (7.9±0.5) cm vs. (8.2±0.5) cm, F= 9.605, P< 0.01, (0.9±0.1) cm and (1.0±0.0) cm vs. (1.1±0.1) cm, F= 71.437, P< 0.01). Besides, according to above data, the right step length and toe distance in HIBI-M group were significantly shorter than those in the HIBI-F group (both P< 0.01). Furthermore, the right step length was significantly shorter than the left step length ((8.3±0.4) and (8.3±0.5) cm, t= 5.289 and 10.580, P= 0.001 and 0.010, respectively) and toe distance ((1.1±0.1) and (1.1±0.1) cm, t= 7.953 and 6.435, respectively, both P <0.01) in both HIBI-M group and HIBI-F group. Similarly, the synaptic gap of the left precentral gyrus neurons was longer in HIBI-M group and HIBI-F group than that in control group ((23.4±1.3) and (19.7±1.6) nm vs. (18.9±0.6) nm, F= 71.719, P <0.01), and also longer in HIBI-M group than that in HIBI-F group ( t= 7.645, P <0.01). Likewise, the residual brain volume in HIBI-M group and HIBI-F group was significantly less than that in control group ((67±4)% and (75±5)% vs. 100%, F= 406.122, P <0.01), and the residual brain volume in HIBI-M group was significantly less than that in HIBI-F group ( t= -5.281, P <0.01). Conclusions: Male neonatal rats are more vulnerable to HIBI and have severer subsequent brain injury and hemiplegia. Different treatment strategies for HIBI patients of different sexes should be developed.

Objective: To investigate the economic burden of hepatitis C patients and related factors in Guangdong province. Methods: In this study, cluster sampling method was used to select cases, including acute hepatitis C, chronic hepatitis C and liver cirrhosis cases from eligible outpatients and inpatients in 1 or 2 large general hospitals in all the 21 cities in Guangdong province. Questionnaire survey was conducted for all the hepatitis C patients to analyze their economic burden, while multivariate linear regression model was used to identify the related influencing factors. Results: A total of 356 hepatitis C patients were enrolled in the study, with 176 outpatients (49.4 % ) and 180 inpatients (50.6 % ) respectively. The average age of the study subjects was (44.79±11.73) year-olds. The annual direct economic costs of patients with acute hepatitis C, chronic hepatitis C and liver cirrhosis were 10 703.22 ( IQR : 7 396.75-16 891.91), 14 886.63 ( IQR : 7 274.00-30 228.25) and 28 874.00 ( IQR : 13 093.69-56 350.00) Yuan (RMB) respectively. The annual indirect costs appeared as 2 426.99 ( IQR : 1 912.18-7 354.52), 3 235.99 ( IQR : 1 323.81-6 619.07) and 5 442.35 ( IQR : 3 235.99-10 296.33) Yuan (RMB) respectively. The annual intangible costs were 5 000.00 ( IQR :2 000.00-10 000.00), 10 000.00 ( IQR : 4 000.00-30 000.00) and 10 000.00 ( IQR : 3 000.00-100 000.00) Yuan (RMB) respectively. The annual total costs were 22 306.17 ( IQR : 14 581.24-50 569.17), 38 050.33 ( IQR : 17 449.57-68 319.62) and 80 152.18 ( IQR : 40 856.09-228 460.79) Yuan (RMB) respectively. Results from the multiple linear regression analysis showed that factors as: annual hospitalization days, annual number of outpatient visits, annual number of hospitalization, type of disease and the levels of the hospitals were related to the economic burden of patients with hepatitis C. Conclusion: Patients with HCV-related diseases presented serious economic problem which calls for close attention in Guangdong province.

Objective: To study the association between maternal pre-pregnancy body mass index (BMI) and adverse outcomes of late preterm infants (LPI)., Methods: A total of 367 LPI who were born from January 2011 to December 2015 and admitted to the neonatal ward were enrolled. The BMI criteria for Chinese population were used to analyze the factors for maternal pre-pregnancy BMI and its association with adverse outcomes of LPI (1 minute Apgar score ≤7, delivery room resuscitation, hospitalization days after birth >7 days, and ventilation duration ≥6 hours)., Results: Of all LPIs, there were 64 LPI (17.4%) in the low maternal pre-pregnancy BMI group, 243 LPI (66.2%) in the normal maternal pre-pregnancy BMI group, and 60 LPI (16.4%) in the high maternal pre-pregnancy BMI group. Low pre-pregnancy BMI was the risk factor for 1 minute Apgar score ≤7 (OR=3.243, 95% CI: 1.102-9.546) and need for delivery room resuscitation (OR=3.492, 95%CI: 1.090-11.190), and high pre-pregnancy BMI was the risk factor for hospitalization days after birth >7 days (OR=1.992, 95%CI: 1.024-3.874)., Conclusions: Abnormal maternal pre-pregnancy BMI has adverse effects on the outcomes of LPI. In order to reduce these adverse outcomes BMI should be controlled within the normal range in pregnant women.

To prepare the hawthorn leaves flavonoids self-microemulsifying membrane controlled-release coated drop pill, and to study its release rate in vitro and pharmacokinetics study in vivo. In order to improve the dissolution of hawthorn leaves flavonoids, self-microemulsifying technology was used to prepare the hawthorn leaves flavonoids self-microemulsion. Hawthorn leaves flavonoids self-microemulsifying drop pill was prepared with the PEG 6000. Studies were made on the in vitro release of flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills and the in vivo pharmacokinetic in rats. The prescription of flavonoids from hawthorn leaves self-micro-emulsifying drop pills was 0.25 g of flavonoids from hawthorn leaves, 0.25 g of iodophenyl maleimide, 0.375 g of polyethylene glycol 400, 0.375 g of cremophor RH 40 and 2 g of polyethylene glycol 6000. The optimized prescription was 4 g of ethyl cellulose 20, 0.64 g of polyethylene glycol 400, 1.8 g of diethyl phthalate, and the weight of coating materials increased by 3.5%. Flavonoids from hawthorn leaves self-micro-emulsifying membrane-moderated coated drop pills complied with the design of sustained-release in 12 h in terms of in vitro release and in vivo pharmacokinetic parameters in rats, and its bioavailability was 2.47 times of quick-release drop pills. Slightly soluble flavonoids from hawthorn leaves could be made into sustained-release preparations by the self-micro-emulsifying and coating technology.

Objective: To explore the effect of human umbilical cord blood mononuclear cells (UCBMC) promoting nerve behavior function and brain tissue recovery of neonatal SD rat with hypoxic ischemic brain injury (HIBI)., Method: A modified newborn rat model that had a combined hypoxic and ischemic brain injury as described by Rice-Vannucci was used, early nervous reflex, the Morris water maze and walking track analysis were used to evaluate nervous behavioral function, and brain MRI, HE staining to evaluate brain damage recovery., Result: Newborn rat Rice-Vannucci model showed significant brain atrophy, obvious hemiplegia of contralateral limbs,e.g right step length [(7.67 ± 0.46) cm vs. (8.22 ± 0.50) cm, F = 1.494] and toe distance [(0.93 ± 0.06) cm vs. (1.12 ± 0.55) cm, F = 0.186] were significantly reduced compared with left side, learning and memory ability was significantly impaired compared with normal control group (P < 0.01); Cliff aversion [(8.44 ± 2.38) s vs.(14.22 ± 5.07) s, t = 4.618] and negative geotaxis reflex time [(7.26 ± 2.00) s vs. (11.76 ± 3.73) s, t = 4.755] on postnatal 14 days of HIBI+ transplantation group were significantly reduced compared with HIBI+NaCl group (P < 0.01) ; the Morris water maze experiment showed escape latency [ (23.11 ± 6.64) s vs. (34.04 ± 12.95) s, t = 3.356] and swimming distance [ (9.12 ± 1.21) cm vs.(12.70 ± 1.53) cm, t = 17.095] of HIBI+transplantation group were significantly reduced compared with those of HIBI+NaCl group (P < 0.01) ; the residual brain volume on postnatal 10 d [ (75.37 ± 4.53)% vs. (67.17 ± 4.08)%, t = -6.017] and 67 d [ (69.05 ± 3.58)% vs.(60.83 ± 3.69)%, t = -7.148]of HIBI+ transplantation group were significantly larger than those of HIBI+NaCl group (P < 0.01); After human UCBMC transplantation, left cortical edema significantly reduced and nerve cell necrosis of HIBI+ transplantation group is not obvious compared with HIBI+NaCl group., Conclusion: Human UCBMC intraperitoneal transplantation significantly promoted recovery of injured brain cells and neurobehavioral function development.

Objective: To evaluate the outcome and indication of the reconstruction of oral and maxillofacial postoperative defects by submental artery island myocutaneous flaps., Methods: Sixty eight cases with the reconstruction of oral and maxillofacial defects by submental artery island myocutaneous flaps from January 2006 to May 2010 were analysed retrospectively. Primary lesions included carcinomas originating from tongue (28 cases), palate (13 cases), mouth floor (9 cases), gingiva (4 cases), buccal mucosa (6 cases), lip (3 cases), and other malignant or benign tumors (5 cases). The ages ranged from 25 to 84 years (mean 58 years); 47 males and 21 females. The sizes of skin paddle varied from a minimum of 4 cm × 4 cm to a maximum of 15 cm × 10 cm., Results: Of the 68 flaps, 62 were survival, 4 had partial necrosis but healed with treatments, and 2 failed due to complete necrosis. Appearance and functions of recipient sites were satisfactory. The followed-up time was 3 - 24 months, local recurrence occurred in 5 cases and cervical lymph node metastases were found in 15 patients., Conclusion: Submental island flap is reliable for the reconstruction of postoperative defects in early oral cancer without regional lymph node metastasis or in benign tumor.

Effects of phosphorus of low concentrations on the growth and the phosphorus removal efficiency of Scenedesmus obliquus were investigated in this study. Results showed that Scenedesmus obliquus achieved a phosphorus removal efficiency of 100% within 22 h when the initial algal cell concentration was 1 x 10(5) /mL and the initial phosphorus concentration was 0.02-0.10 mg/L. With the initial phosphorus concentration increased from 0.02 mg/L to 0.10 mg/L, both growth velocity of Scenedesmus obliquus and maximum biomass increased obviously. Research found that phosphorus concentration had a significant influence on cell morphology of algal. In the external phosphorus sufficient conditions, most of algae cell present as four cells gather round form, then transformed into two cells side by side form in the absence of external phosphorus in culture medium, Finally in single as the main form of existence.

Objective: To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection., Methods: Fresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts., Results: Ameloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05)., Conclusion: The xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.

Objective: To investigate the feasibility of endoscopy-assisted partial-superficial parotidectomy., Methods: 38 cases with benign tumors located in the superficial lobe of the parotid gland were randomly assigned to receive conventional (20 cases) or endoscopic (18 cases) partial-superficial parotidectomy. Two short incisions, which were 2 to approximately 2.5 cm in length and located at retromandibular and postauricular area, were adopted for endoscopy-assisted surgery. The facial nerve was dissected through retrograde approach., Results: The tumors were successfully resected with endoscopy in 18 cases. The operation time was not significantly different between the conventional and endoscopy-assisted procedures (P > 0.05). The intraoperative blood loss was markedly lower in endoscopy-assisted group, compared with conventional group (P < 0.01). All the 18 cases with endoscopy-assisted surgery were satisfactory with the postoperative cosmetic results. The great auricular nerve was preserved very well in 12 patients (66.6%). Transient facial paralysis happened in 1 case and relieved 1 month later. Salivary fistula occurred in 1 case and recovered after dressing with pressure for 2 weeks. All the patients were followed up for 24 to approximately 50 months (mean, 39 months) without relapse., Conclusions: Endoscopy-assisted partial-superficial parotidectomy can successfully treat benign tumors located in the superficial lobe of parotid gland with a better postoperative cosmetic result.

In the present paper, the authors aimed to detect the binding of monoclonal antibody and polyclonal antibodies in sera of patients with recombinant house dust mite allergen (rDer p2) by surface plasmon resonance (SPR) technique. This technique is superior to other methods, like enzyme-linked immunosorbent assays (ELISAs) in studying the interaction between biomolecules because no labeling and sample separation are needed. The allergen rDer p2 was immobilized on carboxymethyldextran-modified sensor chip surface by amine coupling. Surface plasmon resonance measurements of monoclonal antibody and polyclonal antibodies in patients' sera revealed that their bindings diverge widely; the binding of patients' sera was remarkably lower than that of monoclonal antibody. At the same time, the binding of patients' sera with rDer p2 varied among patients allergic to dust mite. This study could provide an easy, fast and real-time way for clinical allergic diseases diagnosis.

Objective: To explore the application of MS-CT and 3D reconstruction in diagnosis and treatment of vascular malformations in head and neck., Methods: 20 cases with vascular malformations in head and neck underwent MS-CT and 3-D reconstruction. Then the treatment was determined based on the results of MSCT scanning. The postoperative results were evaluated., Results: The images of MS-CT showed the edge of vascular malformations partially or completely in 16 cases of venous malformations. The lesion's anatomic site and 3-D position was obtained. The 3-D images also showed the overexpanded supply arteries in 4 case of arteriovenous malformations. 2 case of venous malformations in lip underwent resection and healed completely. 12 cases of venous malformations in buccal and floor of mouth were treated with compartmentalized sclerotherapy with partial lesion involution. 2 case of venous malformations in mouth floor were treated with operation followed by sclerotherpy with partial lesion involution. 4 cases of arteriovenous malformations were treated with Superselective Artery Embolization with partial lesion involution., Conclusion: MS-CT and 3D reconstruction can play an important role in diagnosis and treatment of vascular malformations in head and neck.

Objective: To explore the effect of the patients' articulator function after reconstruction of hemi-tongue defect with forearm flap (FAP) or prime close (PC)., Methods: 36 patients who underwent hemiglossectomy were investigated after radical surgery for TSCC. 20 cases were reconstructed with FAP flaps and 16 with primary closure. The patients' articulator functions were evaluated by articulation tests. VS-9700 was used to analyze the speech character when they pronounce /ji/., Results: 1) The speech articulation of patients who underwent hemi-tongue reconstruction with FAF was better than that of patients with PC, and there was significant difference between them (P < 0.05). 2) The first formant (F1) of /i/ of the PC group was lower than that of the control group (P < 0.05). But the second formant (F2) of the PC group was higher than that of the control group (P < 0.05). The first formant (F1) of /i/ of the FAF group was lower than that of the control group (P < 0.05), but there were no significant differences between FAF group and control group in F2 of /i/ (P > 0.05)., Conclusions: Articulator function can be well achieved by forearm flaps reconstruction to hemi-tongue defect patients.

Objective: To research the treatment effect of a complex prescription of Chinese crude drug in BALB/c athymic mice., Methods: 48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, they were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug of different density as the low, middle, high and FF207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and survial time, volume of tumors, exponent of tumors and the tumor metastasis in livers were observed; and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry., Results: 4 weeks later, the total survival rate in treatment group (A + B + C) was 50% and more higher than the control group (0%) treated by FT207, P < 0.01. The survival rate in group A, B, C was higher than group D, except group A, there was significant differences, P < 0.05 or 0.01. And no differences were observed between the treatment groups and the control group in volume of tumors and exponent of tumors, P > 0.05. Tumor metastasis in livers of the treatment group is less than the control's, P < 0.05. Negative correlation between volume of tumors and survival time, positive correlation between tumors volume and spleen exponent were observed, P < 0.01. The result of immunohistochemistry represented that the intensity of PTEN in latero-cancer tissue was the highest, and then the hepatic tissue, the lowest was cancer tissue, P < 0.01. It also represented that the intensity of PTEN in treatment groups (A, B, C) was more higher than the control group (D), and group B was the highest in the treatment groups, P < 0.05 or P < 0.01. However, there was no significant statistic difference between group A and group C, P > 0.05., Conclusion: This complex prescription of Chinese crude drug can prolong the survival time and decrease tumor metastasis in livers of these experimental mice. Too large tumor volume is an main death cause of model. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

Objective: To evaluate the oncologic efficacy, surgical safety, postoperative beauty, and morbidity of an ideal procedure of modified radical surgery in tongue cancer at early stages., Methods: Six patients with early stage squamous cell carcinoma of tongue underwent a procedure of modified radical surgery, in which an inconspicuous incision was used, the external jugular vein, sternocleidomastoid muscle, spino-accessory nerve, and greater auricular nerve were preserved, half of glosso and mouth floor were excised, and the defect was repaired instantly. Recurrence of glosso, mouth floor and neck, regional edema, shoulder dysfunction, auricle sensibility, oral cavity function and beautiful outlook on the face and neck were evaluated clinically., Results: Compared with classic radical neck dissection, this procedure of modified radical surgery showed an inconspicuous incision and pretty appearance, minor edema on face and neck, better shoulder function, and sensation of auricular skin. No recurred to the glosso, mouth floor, and neck was found during follow-up. The patients showed better oncological safety, pretty appearance of tongue, and better oral function of speech, swallow and mastication., Conclusion: This ideal procedure of modified radical surgery in tongue cancer at early stages lessens or avoids destruction of face and neck, shoulder malfunction, numbness of auricular skin, and oral dysfunction of speech, swallow and mastication without impairment on the oncologic safety of the radical surgery, and improves the quality of life of the patients.

Objective: To study the effects of adramycin to disturb telomeric extention reaction mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form guanine-quadruplex (G4) structures., Methods: In the presence of adriamycin, d(TTAGGG)4, d(TTAGAG)4, d(TTAGGG)5 and d(TTAGGGT) were analyzed by electrophoretic mobility shift assay. The mobility of d(TTAGGG)3, d(TTAGGG)4 and d(TrAGGG)5 in native polyacrylamide electrophoresis were observed. Methylation protection experiments were performed to investigate the effects of adriamycin on methylation of guanine in d(TTAGGG)4 and d(TTAGAG)4. The traditional telomeric repeats amplification protocol (TRAP) and modified TRAP-G4 assays were, respectively, used to analyze the different characteristcs of adriamycin's inhibiting telomeric extension mediated by telomerase of Tca8113 cells., Results: At 5.00 microg/mL of adriamycin, conversion of some of linear d(TrAGGG)4 and d(TrAGGG)5to the new, high-mobility bands formed by complex with special second structures were found in the mobility shift assay. Adriamycin at 1.25 microg/mL protected the G in d(TIAGGG)4 from methylating. Adriamycin at 2.50 microg/mL or 1.25 microg/mL partially inhibited the telomeric extension lengthened by telomerase of Tca8113 cells in TRAP assay, but completely did so in TRAP-G4 assay., Conclusion: Adriamycin is able to disturb telomeric extention mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form intra-molecular G4 structures.

Objective: To research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402., Method: 48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent)., Result: All of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C., Conclusion: The higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

Objective: To investigate the effects of survivin short hairpin RNA (shRNA) on survivin expression, cell apoptosis, and chemosensitivity of human tongue cancer cell Tca8113 to cisplatin., Methods: Survivin-directed shRNA plasmid vector was delivered into Tca8113 cells with lipofectamine(TM) 2000 reagent. Survivin expression was detected with the reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Flow cytometry was used to examine cell apoptosis, and the sensitivity to anticancer agents was evaluated by methyl thiazolyl tetrazolium (MTT) assay., Results: After survivin shRNA vector transfection in Tca8113 cells, the expression of mRNA/protein declined significantly, and the apoptotic rate increased in time-dependent manner up to 37.9% at 48 hours. RNAi-mediated survivin reduction selectively inhibited growth and enhanced chemosensitivity of cisplatin but not of 5-fluorouracil., Conclusions: Survivin shRNA could inhibit the expression of survivin mRNA and it's protein and enhance the chemosensitivity of cisplatin.

Objective: To observe the induction of apoptosis of cisplatin (DDP) to oral squamous cell carcinoma cell line (Tca8113) in vitro and study the role of Survivin on the apoptosis of Tca8113 cells induced by cisplatin., Methods: The inhibitory effects of different doses of DDP on Tca8113 cells were assayed with MTT test. Apoptosis was determined by flow cytometry. The expression of Survivin was detected by RT-PCR and immunocytochemistry., Results: Cisplatin obviously inhibited Tca8113 cells growth in a dose and time dependent manner. The apoptotic index showed the similar trend. Survivin gene expression was decreased with increasing of time and reached the lowest level at 24 hours after DDP treatment, then increased after that time., Conclusion: Cisplatin gene can effectively induce apoptosis in Tca8113 cells and the inhibition of Survivin gene expression may play a critical role on Tca8113 cell apoptosis induced by cisplatin.

Objective: To establish an subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice., Methods: Ameloblastoma cells were absorbed by primary culture, repeat attachment and pancreas proteolytic enzyme were both used to purify them. Then, the purified cells were implanted subcutaneously into the nude mice. The specimens were respectively investigated by microscope in different spots after implanting., Results: Ameloblastoma cells can survive in all of the 8 nude mice. The xenograft can be found on 23 days after implanting. The rate of successful inocutation is 25%. The subcutaneously xenotransplanted tumor cells can be found with microscope in the inter-muscle tissues of nude mice., Conclusion: The subcutaneously xenotransplanted tumor model of human ameloblastoma in nude mice was successfully established and it may benefit to further studies on this tumor.

Recombinant proteins extracted from inclusion body remain in denaturation status. Renaturation in vitro after initial purification is a key step of downstream processing. A common method of renaturation of recombinant proteins is the dilution method. With Bla g 2 as a model protein, the conformational changes of denatured and renatured Bla g 2 were investigated by applying fluorescence spectra. The effects of different urea concentrations, different SDS concentrations and different pH on the fluorescence intensity of renatured protein were also investigated. The reasons for these were studied with the knowledge of molecular structure.

Objective: To construct the eukaryotic expression vector of TIMP-2 gene and to explore its expression in human ameloblastoma cell in vitro., Methods: The aimed gene fragment was obtained by RT-PCR. And then, molecmicrolar cloning technology and enzyme digestion were used to connect the gene with the plasmid PcDNA3.1(+), which can be expressed in eukaryotic cells and a report gene: green fluorescent protein gene (GFP) was already existed in the plasmid. We named the eukaryotic expression vector, which contended our aimed gene TIMP-2 as well as report gene GFP, PcDNA3.1(+)/GFP-TIMP-2. The vector was identified by PCR analysis, EcoR I and Xho I restriction analysis and Sequence analysis. After the PcDNA3.1(+)/GFP-TIMP-2 was transfected into cultured human ameloblastoma cell, RT-PCR and Flow Cytometry (FCM) and Microscope wre respectively performed to evaluate the effect of transfection and expression., Results: The constructed vector PcDNA3.1(+)/GFP-TIMP-2 was proved correct by enzyme digestion and sequencing analysis. After PcDNA3.1(+)/GFP-TIMP-2 was trasnfected into cultured human ameloblastoma cell, the rate of transfection is 47.6% (Analysis report of FCM), the green fluorescence was found in plasm (observed with fluo-microwave), the expression of TIMP-2 mRNA was elevated 2.4 times compared with the control group., Conclusions: PcDNA3.1(+)/GFP-TIMP-2 was successfully constructed and it could be transfected into cultured human ameloblastoma cell. It may be benefit to further study of the relationship between the TIMP-2 gene and the behaviour of ameloblastoma.

Objective: To investigate the role of telomerase and telomere repeat binding factors (TRF) in apoptosis., Methods: The proliferative activity of Tca8113 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. After Tca8113 cells were treated with adriamycin at 5 mg/L, apoptotic morphology was observed under microscope with Giemsa staining and apoptosis examined by flow cytometry; analysis of telomerase activity was performed by TRAP-enzyme-linked immunosorbent assay; expression and expression level of TRF proteins were detected with immunohistochemical staining and immunofluorescence label assay, respectively., Results: After Tca8113 cells were treated with adriamycin at 5 mg/L for 5 days and 7 days, the cells apoptosis was found. Telomerase activity dropped in time-dependent manner. Expression of TRF proteins appeared in nucleus of the cells. No statistical difference in expression levels of TRF was observed between the treated and untreated cells., Conclusions: Tca8113 cells apoptosis induced by adriamycin decreased telomerase activity, but did not influence the expression level of TRF proteins.

Objective: To observe the therapeutic effect of CDglyTK gene mediated by radiation-inducible promoters in the treatment of buccal carcinoma in Golden Hamster., Methods: Animal models of buccal carcinoma in golden hamster were established by painting 0.5% dimethyl-benzanthracene. The plasmids pcDNA (+) 3.1/E-CDglyTK were transfected into tumors by lipofectamine. 24 h later, the tumors were exposed to 3 Gy irradiation. Animals were monitored at regular intervals for volume of tumors. CDglyTK mRNA was assayed by RT-PCR. Apoptosis and proliferating cell nuclear antigen were detected respectively by in situ end-labeling and immunohistochemical methods., Results: Compared with control groups, the tumor was suppressed obviously by CDglyTK gene therapy combined with 3 Gy induction radiation. The expression of CDglyTK gene could be detected by RT-PCR in the transfected tumor, and up-regulation of CDglyTK expression was found in tumor exposed to radiation (P < 0.05). There was significant difference in apoptosis index or proliferation index between tumor without irradiation and tumor with irradiation (P < 0.05)., Conclusions: The radiation-inducible promoter can be served as a molecular switch to regulate the expression of CDglyTK gene in buccal carcinoma in golden hamster, and low dose induction radiation can significantly improve the therapeutic effects.

Objective: To determine whether the human telomerase reverse transcriptase (hTERT) gene silencing could be effectively induced by PCR-derived siRNA expression cassettes (SEC) transfected by the fifth generation polyamidoamine dendrimer (G5 PAMAM-D) in Tca8113 cells., Methods: Four SEC were rationally designed and constructed based on a two-step PCR reaction. The SEC were then transferred into Tca8113 cells using G5 PAMAM-D, and hTERT expression was investigated by real-time fluorescence-quantitative reverse transcriptase-PCR and western blot analysis., Results: The RNA interference effects of the SEC targeted for varying hTERT mRNA positions showed a significant disparity. Among them, SEC-A revealed the most potent inhibitory effects (above 95% of reduction), followed by SEC-D and SEC-C, and SEC-B had no effect on hTERT expression (P > 0.05). That the endogenous hTERT gene silencing induced by G5 PAMAM dendrimer-mediated SEC-A was highly sequence-specific, and multiple transfection as well as properties of the vectors were routinely attributable to the specific suppression., Conclusions: Specific inhibition of endogenous hTERT expression by use of a PCR-based short hairpin siRNA technique and dendrimer transfer system may serve as a novel strategy for treatment of tongue cancers expressing hTERT in vitro.

Objective: To explore the clinical value and safety of using rib-major pectoralis myocutaneous flap carrying costal parietal pleura in combined repair of large soft and hard tissue defect caused by radical surgery of advanced tongue cancer., Methods: Six patients with advanced tongue carcinoma involving the floor of mouth and mandible were performed combined radical neck dissection with glossectomy and mandibulectomy, which caused large soft and hard tissue defect. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura were transferred for immediate repair of the large defects. The rib flaps were applied for the repair of mandible, and the major pectoralis myocutaneous flaps were applied for the reconstruction of tongue and floor of mouth., Results: Six patients recovered well after operation. Six rib-major pectoralis myocutaneous flaps carrying costal parietal pleura survived well; the wounds of surgical incision of the oral cavity, neck, and chest healed up. The reconstructed tongue and the lower face appearance were satisfactory, the occlusion relationships were normal; the speaking as well as swallowing functions recovered., Conclusions: It's safe and reliable to use rib-major pectoralis myocutaneous flap carrying costal parietal pleura to repair large soft and hard tissue defect in oral and maxillofacial region. Opening pleural cavity and harvest costal parietal pleura would not influence patients' thoracic movement and breath function and would not cause other complications. It's simple and safe for harvesting the composite flap. Carrying costal parietal pleura assures the sufficient blood supply of rib in the composite flap.

Objective: To investigate the feasibility and safety of intraoral submandibular gland excision., Methods: Analyze the relationship between the external maxillary artery and submandibular gland, and offer a reliable anatomical base for 10 cases of intraoral submandibular gland excision, including 8 cases of chronic sialadenitis, 1 case of pleomorphic adenoma and 1 case of cyst of submandibular gland., Results: The external maxillary artery went across the surface of gland submandibular, and its branches provided nutrition for the gland in most cases. The results of 10 cases intraoral submandibular gland excision were effective and satisfied, without major complications. The average time of operation was 50 minutes and the average hemorrhage of operation was 60 ml., Conclusion: Intraoral submandibular gland excision is safe and feasible for chronic sialadenitis and cyst of submandibular gland and some of benign tumor submandibular gland as long as indications strictly controlled and the external maxillary artery well coped with.

Objective: To observe the therapeutic effect of CDglyTK gene mediated by synthetic radiation-inducible promoter in the treatment of Tca8113 cells., Methods: CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1 (+) to construct plasmid pcDNA3.1 (+)-CDglyTK, and then the synthetic radiation-inducible promoter in pMD18 -T -E was inserted into pcDNA3.1 (+) -CDglyTK to construct plasmid pcDNA3.1 (+ )/E -CDglyTK. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine, and then exposed to 3 Gy irradiation. Cytotoxicity was evaluated by MTT. The expression of CDglyTK gene was detected by RT-PCR. The apoptosis and proliferation were examined by flow cytomtery., Results: The plasmid pcDNA3.1 (+)/E-CDglyTK was constructed successfully. The comparative survival rate of Tca8113 cells was markedly decreased by induction irradiation. Up-regulation of CDglyTK expression was found in Tca8113 cells exposed to irradiation. The apoptosis index (AI) of Tca8113 cells exposed to irradiation was higher than that of Tca8113 cells without irradiation, the other way round, the proliferation index (PI) of Tca8113 cells exposed to irradiation was lower than that of Tca8113 cells without irradiation., Conclusion: The synthetic radiation-inducible promoter can be served as a molecular switch to improve the expression of CDglyTK gene in Tca8113 cells, and low dose induction radiation can significantly improve the therapeutic efficiency.

Background & Objective: The intensity of lymphatic metastasis consists of lymph node metastasis number (LMN) and lymph node metastasis ratio (LMR). LMR is the ratio of positive nodes to dissected nodes. LMN and LMR are 2 important prognostic factors of esophageal cancer, and are adopted in tumor staging. This study was to assess the lymphatic metastasis intensity of thoracic esophageal squamous cell carcinoma (ESCC), and explore the influential factors and lymphadenectomy pattern., Methods: A total of 120 patients with ESCC had been operated in the Second Affiliated Hospital of Sun Yat-sen University from 1998 to 2000. The lymph nodes were dissected according to the mapping scheme of the American Thoracic Society (ATS) modified by Casson et al., Results: A total of 2 631 lymph nodes were dissected with an average of 22 lymph nodes in each case. The cervical LMR was significantly higher in the upper thoracic ESCC than in the middle and lower thoracic ESCC (20.9% vs. 12.9% and 6.8%, P<0.05). The left gastric LMR was significantly higher in the lower thoracic ESCC than in the middle and upper thoracic ESCC (37.5% vs. 17.5% and 7.1%, P<0.05). Subcarinal metastatic lymph nodes were often found in the middle thoracic ESCC. T stage, histological differentiation, and circum wall involvement degree were correlated to lymphatic metastasis intensity (P<0.05); the length of esophageal lesion had no correlation to lymphatic metastasis intensity (P>0.05). The survival time of the patients received the right thoracic esophagectomy with 3-field lymphadenectomy (3-FL) was significantly longer than that of the patients received the left thoracic esophagectomy with 2-FL (P<0.05)., Conclusions: During the operation on ESCC, the regions with high lymphatic metastasis intensity should be dissected. T stage, histological differentiation, and circum wall involvement degree are important influential factors of lymphatic metastasis intensity. The right thoracic esophagectomy with 3-FL is superior to the left thoracic esophagectomy with 2-FL.

The recombinant allergen, Bla g 2, was expressed by prokaryotic vector E. coli and eukaryotic vector P. Pastoris. The different structures and configurations of the Bla g 2 from E. coli and P. Pastoris were studied by fluorescence and circular dichroism. The secondary structures of Bla g 2 in solution, and the composition besides the type of its tertiary structure were proposed. These studies help understand the differences between prokaryotic and eukaryotic expression systems, reveal the relationship between the structure and the function of Bla g 2, and improve the production of this significant allergen.

Objective: To investigate the relationship between matrix metalloproteinase-2 (MMP-2)activity and cell proliferation, growth and invasion of ameloblastoma., Methods: The cells and xenograft of ameloblastoma were treated with MMP-2 inhibitor Ro31-9790 and the effects of Ro31-9790 on the cell proliferation and growth of ameloblastoma were observed. Primary culture in vitro, subcapsular kidney xenograft in vivo, MTT assay, flow cytometry, neoplastic volume measurement and histochemistry were employed to study the effects of cell proliferation and growth produced by Ro31-9790., Results: There was no significant different in cell proliferation at same interval among several groups (P > 0.05). The ratio of G0/G1 stage, G2/M stage and apoptotic cells didn't increase following increased Ro31-9790, and the ratio of S stage cells also didn't reduce following increased Ro31-9790. The tumor volume and its increase in treatment group were significant less than those in control group., Conclusion: Ro31-9790 does not influence proliferation of ameloblastoma cells in vitro, but it can effectively inhibit the ameloblastoma growth in vivo. MMP-2 activity has no relationship to proliferation of ameloblastoma cells, but it can contribute to the ameloblastoma growth and may be a reason of invasion in ameloblastoma.

Two new MAR segments (M14 and M17) were cloned from tobacco genome. Both of the sequences contained several typical consensus sequences of MARs, which were different from the original MAR sequence, such as 90%AT-box, A-box, T-box, the base unpairing regions (BUR), autonomously replicating sequences (ARS), the consensus sequence for topoisomerase II, MAR recognition sequence (MRS), origin of replication (ORI), curved DNA motifs and ATATTT et al. To investigate the effects of these two sequences on gene expression in transgenic plants, 3 plant expression vectors were constructed with uidA gene coding beta-glucuronidase (GUS) which were flanked on one side and on both sides by the MARs we obtained. These plant expression vectors with one or two MARs were transformed into tobaccos via Agrobacterium-mediated transformation method, with the plant expression vector pCAMBIA2301 without MAR and wild type tobacco as controls. GUS histochemical staining results showed that the uidA gene expressed stably in transgenic tobaccos. Quantitative detection of GUS activity showed that the MARs could increase GUS expression levels in vivo in contrast to the controls, wherever they were flanked on one side or both sides of uidA gene. The vector ligated with MARs in the same direction on both sides of uidA could increase the GUS expression level much better than both vectors which just ligated with single MARs on one side. The former one increased the average GUS activity for 3.14 folds, but 1.56 and 2.43 folds for the latter two vectors with single MARs respectively contrasting to the pCAMBIA2301 control. But the expression differences among individual transformants were still obvious. Therefore, it was concluded that the DNA sequences we obtained in this experiment were two novel MARs and could enhance gene expression in vivo. In the meanwhile, although the numbers of the MARs typical motifs in M14 were more than in M17, especially the 90% AT box which had been considered to be the highest correlative motif with binding strength in vitro, the enhancement of gene expression was lower yet, which implied no correlation between improvement of gene expression and binding strength between MARs and nuclear matrix in vitro.

Objective: To form ACC-M-GFP cells by transfecting pEGFP-1 into ACC-M cells, and to build up lung metastasis model of adenoid cystic carcinoma for dynamic observation in simulated lung environment., Methods: pEGFP-1 was prepared and then transfected into ACC-M cell lines by using cationic lipid-based gene transfer technique. After successive selection, the ACC-M-GFP cells were collected and inoculated in BALB/C mice. The simulated lung environment was built up, and dynamic observation was performed by laser confocal microscopy within four weeks after transfection., Results: The amount of fluorescent tumor cell colonies and the intensity of fluorescence gradually increased from the second week to the forth week after transfection. In the meantime, only a very small number of tumor cells had the ability to form clones., Conclusions: We successfully built up a lung metastasis model of adenoid cystic carcinoma for dynamic observation. The model is suitable for capturing and analyzing metastatic tumor cells in early stage.

Objective: To detect the role of Vascular endothelial growth factor C (VEGF-C) in the interaction of tongue squamous cell carcinoma (TSCC) with the peri-carcinoma lymphatics., Methods: TSCC cells were implanted on the chorioallantoic membrane (CAM), in which, group A was transfected by plasmid pREP7 including VEGF-C, group B was transfected by plasmid pREP7, and group C was Tca8113. Lymphatic vessels were stained by 5'-Nase. Morphologic analysis was used to evaluate the alteration of lymphatic vessels density (LVD), area and perimeter., Results: The transplanted tumor well on CAM. The expression of VEGF-C in group A was higher than that in control group B and C. The LVD, perimeter and area in group A were (5.3 +/- 0.41)/high-power field, (148 +/- 21) microm and (76.8 +/- 13.5) microm(2) respectively in group A, which were significantly higher than that of group B and C (P < 0.01), while there were no significant difference between group B and C., Conclusions: CAM is ideal model for the research of lymphatic vessels; Over-expression of VEGF-C could induce the dilatation and LVD increase of peri-tumor lymphatic vessels, which maybe one of the mechanisms that VEGF-C metastasis of TSCC through lymphatic vessels.

Objective: To evaluate an island pectoralis major myocutaneous flap for immediate reconstruction of the total tongue body and the floor of the mouth after radical resection of bilateral tongue carcinoma involved., Methods: Form October 2000 to December 2002, seven patients with bilateral tongue carcinoma were selected to treat with salvage surgery, and an island pectoralis major myocutaneous flap were applied for immediate reconstruction of total body of tongue and floor of mouth., Results: Six flaps survived well. Only one flap showed a partial necrosis, with no submandibular fistula, infection, and other complications. The reconstructed tongue and floor of mouth worked well. The speaking and swallowing functions were satisfactory after 2 approximately 16 months' follow-ups. Only one patient died of lung metastasis carcinoma in 9 months after the operation., Conclusions: The island pectoralis major myocutaneous flap could be an ideal selection for the immediate reconstruction of the total tongue body and the floor of mouth.

Objective: To investigate the relationship of Fas mRNA and protein expression and apoptosis in human oral squamous cell carcinoma., Methods: Northern blot and flow cytometry (TUNEL method) were used to detect the expression of Fas mRNA and Fas protein, cell cycle and apoptotic level in oral squamous cell carcinoma. The relationship between Fas gene expression and OSCC apoptosis was analyzed statistically., Results: Fas mRNA and protein could be detected in all five normal oral mucosa specimens. There was positive correlation between expression of Fas mRNA/protein and cell differentiation as well as apoptosis in OSCC (P < 0.005)., Conclusion: The expression of Fas gene was highly correlated with the differentiation and apoptosis in OSCC.

Objective: To investigate the expression of transfected human endostatin (hES) gene in tongue squamous cell carcinoma (TSCC) and its inhibitory effects on the growth of tumor cells in vivo., Methods: Lipofectamine-mediated hES gene was transferred into Tca8113 cells, selected with Blasticidin S; The stable transfected cells were inoculated in BALB/c mice, and then the growth of xenografts was observed. The hES and vascular endothelial growth factor (VEGF) protein expression of xenografts was detected by S-P immuno-histochemical assay. We also detected the microvessel density (MVD) of xenografts with Weidern's method and apoptotic index of the tumor cells by flow cytometry (FCM)., Results: The hES protein expression of xenografts in experimental group was significantly higher than that in control group (P < 0.01), while the expression of VEGF protein was on the other way round (P < 0.01). MVD counting of xenografts in experimental group was lower than that in control group (P < 0.01). The mean apoptotic level of the tumor cells in control group was also lower than in experimental group (P < 0.01). In addition, the inhibitory rate to growth of xenografts induced by hES transfection was 78.9%., Conclusions: hES gene can be transferred into TSCC cells and then induce corresponding protein expression efficiently in xenograft model, resulting in significantly inhibitory effects on the xenografts in vivo.

Objective: To investigate the effect of fas gene transfection and monoclonal anti-fas antibody on tumorigenicity and proliferation of transplanted tumor of Tca8113 cell., Methods: Plasmid including fas gene was transfected into Tca8113 cell by lipofectamine kit. Some transfected cells were treated by monoclonal anti-fas antibody after 48 hours since transfection. Untransfected cell (control), fas-tansfected cell and fas-transfected cell treated with antibody were transplanted to nude mice subcutaneously. Growth of transplanted tumor was observed and recorded regularly. Animals were sacrificed and tumor samples were harvested at the end of experiment. Fas expression in each neoplasm was assessed by RT-PCR. Apoptosis, proliferation and expression of fas protein in tumor tissue were measured by flow cytometry (FCM)., Results: Tumor occurred much later in fas-transfected group and fas-transfected plus antibody treated group. Growth arrest was found in them. RT-PCR and FCM suggested that fas-transfection up-regulated the expression of fas mRNA and protein, increased apoptosis index (AI). But no effect on proliferation index (PI) was observed. Monoclonal anti-fas antibody did not effect the expression of fas mRNA and protein, but increased AI and decreased PI., Conclusion: Fas-transfection suppressing tumorigenesis of Tca8113 cell transplanted in nude mice might be caused by up-regulation of expression of fas gene and enhancement of apoptosis. However, anti-fas antibody suppressing tumorigenesis might be associated with activation of apoptosis and repression of proliferation.

Objective: The purpose of this study was to establish transfergeneic Tca8113 cell and evaluate the expression of human endostatin (hES) gene in the cell colone in vitro., Methods: To transfect hES gene into Tca8113 cells, lipofectamin was complexed with plasmid encoding hES gene, and blasticidin S antibiotic was adopted to select Tca8113--hES cell clone. Immunohistochemistry S-P method was adopted to detect the expression of hES in the transfergenic Tca8113 cell in vitro., Results: Transfected by hES, the transfergenic Tca8113 cells could grow and proliferate in RPMI--1640 culture medium containing blasticidin S antibiotic. The expression rate of hES reached 100%., Conclusion: hES gene can express in hES-transfected Tca8113 cell in vitro.

Background & Objective: Studies have revealed that overexpression of p53 protein has close relation to induction of drug resistance in cancer patients and it can be used as a predictor for chemosensitivity of tumor. Recently, it has been found that specific p53 antibody (p53-Ab) presented in the serum of cancer patients with p53 protein overexpression. Furthermore, the presence of serum p53-Ab is closely correlated with p53 protein overexpression. Thus, serum p53-Ab could theoretically be useful in predicting chemosensitivity in cancer patients. In the present study, we investigated whether preoperative serum p53-Ab is correlated with the postoperative chemosensitivity of esophageal cancer, which was analyzed using in vitro drug response assay., Method: Serum analysis of p53 antibodies was performed by enzyme-linked immunosorbent assay (ELISA) in 38 patients with esophageal cancer preoperatively, then surgically resected specimens were analyzed for their chemosensitivity to cisdichlorodiammineplatinum (DDP),5-fluorouracil (5-FU), and Adriamycin (ADM) by in vitro drug response assay MTT colorimetry., Results: Serum p53-Ab was present in 47.3% of the patients with esophageal cancer. The positive rate of serum p53-Abs in patients was significantly different from that in healthy blood donors (P< 0.01). The positive rate of serum p53-Ab was 25%(5/20) for the patients at stage I and II, and 72.2%(13/18) for the patients at stage III and IV.A significant correlation between serum p53-Ab positive rate and TNM stage was observed (P< 0.05). The positive rate of serum p53-Ab were 21.5%(3/14), 50%(6/12), 75% (9/12) for the patients with well, moderately,and poorly differentiated tumors, respectively. There was significant correlation between serum p53-Ab positive rate and tumor differentiation (P< 0.05). The chemosensitivity to DDP, 5-FU, and ADM (11.1%,16.1%,and 16.1%, respectively) of the patients with positive serum p53-Ab was significantly lower than that of the patients with negative p53-Ab (60%, 45%, and 35%; P< 0.01,P< 0.05, and P< 0.05, respectively)., Conclusion: p53-Ab is not only a prognostic marker for patients with esophageal cancer, but also a predictor of their response to chemotherapy.

Background & Objective: Radical operation of tongue carcinoma causes hemi-tongue defective, which seriously affected the quality of life in these patients. The primary tongue reconstruction guaranteed the healing of the surgical incision at early stage. This may help to recover swallowing and speaking of the patients. However, the vascular crisis happened in the free flap proved to be an obstacle to the extensive application in the tongue reconstruction. This paper illustrated some modifications in the application of the free forearm flap, which may ensure the success in operation., Methods: Some methods like skin flap design, preparation and vascular anastomosis were modified in the application among the 32 tongue carcinoma patients., Results: The surgical incisions of the oral cavity and neck healed up, with no salivary fistula, chylorrhea, submandibular fistula, infection and other complications. After the operations, 6 flaps developed vascular crisis, and 5 flaps were successfully salvaged, meanwhile, one case failed, which resulted in the flap abandonment. The total successful transplantation rate reached 96.9%. The reconstructed tongue recovered, and the speaking as well as swallowing functions re-obtained., Conclusion: The modified methods of tongue reconstruction with free forearm flap improve the clinical effects of reconstructed tongue.

Background & Objective: Abnormal expression of p15 and p16 were commonly found in many kinds of primary tumors, but the possible correlation between p15 and p16 abnormalities and tongue neoplasms is still unknown. This study was designed to investigate the expression of p15 and p16 proteins and their possible correlation with clinicopathology and prognosis of tongue squamous cell carcinoma (TSCC)., Methods: The expression of p15 and p16 were detected by immunohistochemistry (ultra-sensitive SP method) and the results were analyzed quantitatively by imagine cytometry in 45 cases of TSCC and 10 cases of normal tongue tissues, thereafter, the results were analyzed with clinicopathological parameters and survival time., Results: Both p15 and p16 proteins were expressed in squamous epithelia of the normal tongue tissues. The negative expression rates of p15 and p16 protein in TSCC were 46.7%(21/45) and 66.7%(30/45), respectively. 80%(12/15) of the p16 positive cases accompanied with p15 positive staining. However, 85.7%(18/21) of the cases with p15 deletion exhibited p16 negative expression. Most of the cases with both p15 and p16 co-deletion were found in stage III and IV (16/18). The expression rates of p15 and/or p16 in stage I, II were significantly higher than those in stage III, IV (P< 0.05), while p15 and/or p16 expression rates in node-positive groups were much lower than those in node-negative groups (P< 0.01). p15 deletion, or both p15 and p16 co-deletion group, also had a decreased 3-year survival rate (P< 0.05)., Conclusion: The expression of p15 and p16 protein are closely associated with clinical stages and cervical lymph node metastasis of TSCC. p15 deletion, or both p15 and p16 co-deletion in TSCC can also predict a poor prognosis. p15 and p16 expression can be used as parameters for evaluating clinical stages and metastatic potential as well as prognosis of TSCC.

Objective: To describe a hospital outbreak of severe acute respiratory syndrome (SARS) and summarize the clinical features and therapeutic approaches., Methods: Clinical data in this cohort were collected prospectively as they were identified., Results: The outbreak started with a SARS patient from the community on 30 January 2003, followed by a total of 96 people [76 women and 20 men; mean age (29.5 +/- 10.3) years; 93.8% of whom were health care workers] infected in a short period of time after their exposure to this source patient. The incubation period ranged from 1 to 20 days, with a mean of (5.9 +/- 3.5) days. The initial temperature was (38.3 +/- 0.6) degrees C, while the highest was (39.2 +/- 0.6) degrees C (P < 0.001), with a mean fever duration of (9.0 +/- 4.2) days. Other common symptoms included fatigue, cough, mild sputum production, chills, headache, general malaise and myalgia. The radiographic changes were predominantly bilateral and in the middle or lower lung zones. Leukopenia was observed in 67.7% of this cohort. The mean lowest oxygen saturation was (94.8 +/- 3.1)% with supplementary oxygen through a nasal cannula. 68.8% of the patients were treated with methylprednisolone for a mean period l of (4.9 +/- 2.4) days. The initial dose was (67.3 +/- 28.2) mg/d and the maximal dose was (82.4 +/- 30.5) mg/d. Ninety-five patients (99.0%) had a complete clinical recovery, and 1 patient died of progressive acute respiratory distress syndrome. The mean hospitalized duration was (17.2 +/- 8.0) days., Conclusion: SARS appears to be highly contagious and potentially lethal among health care workers, characterized by acute onset and rapid progression. Corticosteroids, antibiotics, human gamma-globulin, interferon-alpha, and antiviral drugs, although used empirically, might be of some benefits in shortening the clinical course.