The cellular sources of leukoregulin (LR) and lymphotoxin (LT) from human peripheral blood leukocytes or spleen cells were examined The ratios of LT activity to LR activity in lymphokine preparations from different individuals were not constant but varied over 640-fold, suggesting that LR and LT activities were mediated by distinctly different entities.Purified LR was used to examine the relative susceptibilities of human or mice cells to LR.Most human tumor cells were very sensitive to LR while human normal cells and mice normal or tumor cells were less sensitive or resistant.Crude preparations of human interferon (Hu IFN)-? or Hu IFN-? were more cytotoxic to human tumor cells than partially purified natural Hu IFN-? or highly purified rHu IFN-?D, and crude or purified LR were more cytotoxic to human tumor cells than Hu IFN-? Hu IFN-?.Hu IFN-? activity but not cytotoxic activity could be removed from preparatons containing Hu IFN-?,LR and LT activities with the aid of monoclonal antibody to Hu IFN-?. Natural killer cytotoxic factor (NKCF) supernatants from the co-culture of human spleen cells and NK susceptible cell line K562 or Molt-4 were strongly cytotoxic to various tumor cell lines.The NKCF activity was completely adsorbed after incubation with Molt-4 cells and completely inactivated by incubating at pH2 for 24 h.The putative receptor for LR or LT was studied using adsorption assay.The present study reveals that human LR activity can be adsorbed from crude LR supernatants by incubation with native or formalin-treated K562 or SMMC-7721 cells at 37℃ or 4℃; LT activity can be adsorbed with L929 cells.Therefore, LR appears to be a cytotoxic/cytostatic factor that is distinct from LT, 1FN and NKCF.

In order to provide highly purified preparation of human ieukoregulin (LR) for the exploration of its anticancer mschatiism, crude preparations of LR and lymphotoxin(LT) derived from human spleen cell cultures induced with phytohem-agglutinin and diterp;ne ester l2-O-tetradecanoylphorbol-i3-acetate were purified by using four purification nmhois : (a) Con A-Sepharose 48 affinity chromatography, by which crude human LR or LT was purified to maximum specific activity of 85 333 U/mg protein or 13333 U/mg protein with a recovery of 28.1% or 31.9%, respectively 5 (b) DEAE-Sjphadex A-50 ion-exchange chromatography, by which partially purified LR or LT obtained after Con A-Sepharose 4B affinity chromatography was further purified 2.44-fold or 3.8o-fold with a recovery of 96% or 92%, respectively ; (c) DEAE-22-celluiose ion-exchange chromatography, by which crude human LR or LT was purified to maximum specific activity of 11 294 U/mg protein or 13 176 U/mg protein with a recovery of 87.5% or 89.6%.respectively ; (d) Blue-Sepharose CL-6B affinity chromatography, by which crude human LR or LT was purified to maximum specific activity of 51.89 U/mg protein or 2594 U/mg protein with a recovery of 53.75% or 5t3.G7%, respectively.Our data indicate that Con A-Sepharose 4B affinity chromatography, DEAE-Sephadex A-50 ion-exchange chromatography.DEAE-22-cellulose ion-exchange chromatography and Blue-Sepharose CL-6B affinity chromatography may be useful in the purification of LR or LT.

A simple, rapid and accurate method is described for the determination of icariin in Fuchun Tablets by reversed-phase high-efficiency liquid chromatography. The samples were first extracted with 70% alcohol for 50 minutes The resulting solutions were chromatographed on aYWG-C18 column.By using a mobile phase of methanoltetrahydrofuran-water (26.5:15.9:56.7) and a detection wavelength at 274 nm, the recovery for 101.02%, CV=2.4%,are obtained.The method can be employed for the quality control of Fuchun Tablets.

Object To optimize the best technical parameters through controlling the different factors. Methods Taking spherical degree and pill weight as index, the above factors were observed by orthogonal test. Results PEG4000 as matrix, methyl silicon oil as refrigerant. Internal and external diameter of burette are 4 1 mm and 6 1 mm, dropping into the refrigerant that composed of 40 ℃—50 ℃,10 ℃—30 ℃,0 ℃—4 ℃ by 20 ℃—30 ℃ droppings per minute. Conclusion The highest finished product with good quality can be got through this process.

Objective: To observe the effect of electroacupuncture (EA) at Lower He-Sea points on the expression levels of interleukin-1β (IL-1β) in the serum and gallbladder tissues, and nuclear factor-κB (NF-κB) in gallbladder tissues of the guinea pigs with acute cholecystitis (AC), and to explore whether Yanglingquan (GB 34), the Lower He-Sea point pertaining to Dan Fu (gallbladder), is relatively specific for the Dan Fu (gallbladder) disorders. Methods: Eighty-two healthy guinea pigs were randomly divided into 6 groups according to the random number table method, a blank group, a model group, a Yanglingquan (GB 34) group, a Zusanli (ST 36) group, a Shangjuxu (ST 37) group, and a Xiajuxu (ST 39) group, with 12 guinea pigs in the blank group while 14 in the other groups, respectively, half males and half females in each group. Except for the blank group, guinea pigs in the other groups were injected with E. coli into the gallbladder to establish AC models. Guinea pigs in the blank group were fed routinely without special treatment; those in the model group were daily tied up for 30 min without EA treatment; those in the 4 groups receiving EA treatment were acupunctured at the corresponding Lower He-Sea points after daily binding and stimulated with the SDZ-V EA instrument. After successful modeling and treatment for 5 d, blood was collected from the abdominal aorta of the guinea pigs, and the gallbladder tissues in each group were isolated for hematoxylin-eosin (HE) staining to observe the morphological changes. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum IL-1β level, and immunohistochemistry (IHC) was used to detect the expression levels of NF-κB and IL-1β in gallbladder. Results: On the 3rd day after modeling, the guinea pigs in the five groups with modeling were mentally depressed with decreased appetite, significantly reduced activities, slouch, lassitude, slack and matted fur, and loose stools; two guinea pigs were selected from each group (one male and one female, not included in the final statistics) to isolate the gallbladder after sacrifice; macroscopic observation showed that the gallbladder wall was differently thickened; the bile color was dark green and opaque with particles suspended or accumulated; light microscope observation showed that the submucosal blood vessels of the gallbladder were congested, along with mucosal edema, ulceration, necrosis, shedding, and a large number of inflammatory cells infiltrating in the lamina propria, indicating that the AC model was successfully prepared. Compared with the model group, the gallbladder tissue injuries of the four groups receiving EA treatment were all differently repaired, the serum IL-1β levels were reduced (P<0.01 or P<0.05), and the IL-1β levels in the gallbladder tissues were reduced (P<0.01 or P<0.05). Compared with the model group, the NF-κB expression level in the Yanglingquan (GB 34) group was significantly reduced (P<0.01), but was not statistical different in the Zusanli (ST 36) group, Shangjuxu (ST 37) group and Xiajuxu (ST 39) group (all P>0.05). Compared with the Yanglingquan (GB 34) group, the gallbladder tissues of the Zusanli (ST 36) group, Shangjuxu (ST 37) group and Xiajuxu (ST 39) group were more severely damaged, and the expression levels of serum IL-1β, the NF-κB and IL-1β in the gallbladder tissues were increased (P<0.01 or P<0.05). Intervention effect of Yanglingquan (GB 34) on AC guinea pigs was superior to that of Zusanli (ST 36), Shangjuxu (ST 37) and Xiajuxu (ST 39). Conclusion: EA at the Lower He-Sea points of the stomach, large intestine, small intestine and gallbladder can produce curative effects on AC guinea pigs and reduced the inflammatory symptoms. Intervention effect of Yanglingquan (GB 34) on AC guinea pigs is superior to that of Zusanli (ST 36), Shangjuxu (ST 37) and Xiajuxu (ST 39). The mechanism of EA at Yanglingquan (GB 34) in treating AC may be regulating IL-1β and NF-κB to control the inflammatory response and improve the gallbladder tissue damage.

Humans ; Mastoiditis ; diagnosis ; surgery ; Otitis Media ; diagnosis ; surgery

Objective:To develope a method for the separation and preparation ginkgolic acid ? 1. Methods: Ginkgolic acids were extracted by alcohol, and adsorbed by silica gel to remove most of the polar impurities. It was preparated with RP HPLC. Results: The extraction ratio of ginkgolic acid ? 1 by this extraction process was above 84%, and the purity of ginkgolic acid ? 1 reached 98%. Conclusion: This procedure is simple and relatively effective.

Objective To investigate the alterations and molecular mechanism of neural cell apoptosis after frontal lobe sharp injury in infant rats.Methods Brain damage was induced by frontal lobe sharp injury.dUTP nick end labeling(TUNEL) method was used to detect cell apoptosis after brain injury,and immunohistochemistry was used to detect the expression of Bax,Bcl-2 proteins in and around the area of injury,respectively.Results The apoptosis cells were found as early as 1 hour and reached a peak at 24 hours post-injury,then gradually diminished until 120 hours when only few apoptosis cells presented.The expression of Bax and Bcl-2 proteins increased significantly 1 hour after frontal lobe injury.Bax protein reached its peak at 12 hours while Bcl-2 protein reached its peak at 1 hour post-injury.The ratio of Bax to Bcl-2 increased after lesion with peak at 24 hours,and then decreased thereafter.At 120 hours,very weak expression of Bax and Bcl-2 protein detected.There were few Bax and Bcl-2 positive cells in frontal lobe cortex in blank control and sham operation group.Conclusions Cell apoptosis exists after frontal lobe sharp injury in infant rats and the change of the number of apoptotic cells is correlated with time course of post-injury.The ratio of Bax to Bcl-2 may play a vital role in cell apoptosis.

Purpose:To investigate prognostic effects of ex pressions of cyclooxygenase-2(COX-2) and epidermal growth factor receptor(EGFR )in non-small-cell lung cancer(NSCLC).Methods:Expression of COX-2 and EGFR in the resected tumors of 60 patients with NSCLC were detected with the method of immunohistochemistry st aining.We used ?2 test and COX regression analysis to compare difference in e xpressions of COX-2 and EGFR in differentiation grade of squamous and adeno-ca rcinoma cells of the lungs and their relationships with the patients survival in NSCLC.Results:The positive rates for COX-2 were 73% and 33% in adeno carcinoma and squamous carcinoma,respectively.The expression rate was significan tly higher in the adenocarcinoma than in the squamous carcinoma.Expressions of C OX-2 and EGFR in NSCLC were not related to patientsage and sex,differentiation grade of tumor cells,TNM staging,size of promary tumor and lymph nodes metastasi s.Median survival time was 30 months and 16 months in COX-2 expression levels o f “+” and of “++”,34 months and 15 months in EGFR expression levels of “+” and of “++” and was significantly lower than in those without expression of C OX-2 or EGFR(P

Objective To discuss appropriate strategy about treatment of rotatory fixation or dislocation of atlantoaxial articulation in children. Methods 32 cases of rotatory fixation or dislocation of atlantoaxial articulation in children were analyzed retrospectively from January 2000 to March 2005, 24 males and 8 females; patients' age was from 3 to 12 years old with an average of 5.5 years old. The mean course of diseases was 6.5 weeks (range: 2 days to 50 weeks). According to Fielding-Hawkins clinical classification, typeⅠ17 cases(1 case with os odontoideum), type Ⅱ 12 cases(1 case with os odontoideum and 1 case with congenital absence of the arch of posterior atlas), type Ⅲ 3 cases. 26 cases received conservational treatments which were recommended as 2-3 weeks of cervical traction, traction weight should be controlled to 1.0-1.5 kg. After a successful reduction a proper external fixation was required to maintain reduction. And another 6 serious cases received posterior atlantoaxial or occipitocervical autograft fusion. Indications for posterior atlantoaxial or occipitocervical atuograft fusion includes: 1) have difficulty in reduction even under proper traction; 2) with obvious neurological symptoms; 3) combined with compensatory deformity of atlantoccipital articulation or other occiptocervical deformities. Results All of 32 cases were followed up from 3 to 50 months with an average of 32 months. 26 cases who received cervical traction and external fixation resulted in satisfactory outcome in which all the torticollis were rectified, bilateral masses were symmetrical on AP position, and ADI was less than 4 mm in dynamic extension and flexation lateral view. ROM in rotation and extension and flexion were completely recovered. All the 6 cases surgically treated obtained sound bony fusion and neural symptoms were improved obviously and torticollis were rectified completely in 4 cases and others (2 cases) stopped progressing after operation. Conclusion Conservative treatment including Glission traction, cranial traction, collar, cast or cervicothoracic external fixation has been proved to be very effective in most of rotatory fixation or dislocation of atlantoaxial articulation in children; however, operative treatment should be considered in the following situations: patients with difficulty in reduction, with neural involvement and compensatory deformity of atlantoccipital articulation or other occipitocervical deformities.