Tumor microenvironment responsive liposomes blocking CXCL12/CXCR4 pathway and synergistically enhancing immune efficacy of anti-PD-L1 / 药学学报

Blocking immune checkpoint programmed cell death receptor 1 (PD-1) or programmed death receptor-ligand 1 (PD-L1) can enhance anti-tumor activity of effector T cells. However, the lack of response in many patients to PD-1/PD-L1 therapy remains a question. Improving the immunosuppressive tumor microenvironment (TME) to enhance the efficacy of immune checkpoint inhibitors has become a promising cancer treatment strategy. We constructed a liposome system (PD-L1/siCXCL12-Lp) of CXCL12 siRNA and anti-PD-L1 peptide with matrix metalloproteinases (MMPs) responsiveness, which combined the TME regulation of siCXCL12 and the immune regulation of anti-PD-L1 peptide. All animal experiments were approved by the Biomedical Ethics Committee of Peking University. The authors found that PD-L1/siCXCL12-Lp directly down-regulated the expression of CXCL12 in vitro (33.8%) and in vivo (15.5%). It also effectively increased the ratio of CD8+/Treg by 20.0%, which helped the anti-PD-L1 peptide to better exert its immune effect. The combination therapy significantly inhibited tumor growth (52.08%) with great safety, which explored a new idea for cancer immunotherapy.

Effects of Chronic Restraint Stress on Apoptosis of Amygdala Cells in Rats / 法医学杂志

OBJECTIVES@#To explore the damage effects of chronic restraint stress (CRS) on amygdala cells through the rat CRS model.@*METHODS@#The rat CRS model was established, and the changes in body weight and adrenal mass in control group and CRS group were monitored at 1 d, 7 d, 14 d and 21 d. The behavior changes were evaluated by the percentage of retention time of open arms and open arm entries using the elevated plus maze (EPM). ELISA was used to detect the concentrations of rat's corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol. The changes of expression of glucocorticoid receptor (GR) and glial fibrillary acidic protein (GFAP) in amygdala were determined by immunohistochemistry and Western blotting. Ultrastructure changes of glial cell were observed by transmission electron microscopy. The apoptosis rate of amygdala was measured by flow cytometry.@*RESULTS@#Compared with the control group at the same time points, body weight of CRS 1 d, 7 d, 14 d and 21 d groups increased slowly, but adrenal mass increased significantly; the serum level of CRH, cortisol and ACTH increased significantly at 7 d, 14 d and 21 d respectively; the expression of GR in amygdala was increased while that of GFAP was decreased; EPM test suggested that the percentage of retention time of open arms and open arm entries decreased significantly after 14 d. The CRS group showed different degrees of glial cell damage in amygdala, and the apoptosis rate of glial cell was significantly increased in 21 d group.@*CONCLUSIONS@#This study successfully established a CRS model in rats, and anxiety-like behavioral changes in model rats may be caused by apoptosis of amygdala astrocytes.

Effect of Huanglian Wendantang on Inflammatory Response and Hepatocyte Burn Death in IGT Rats / 中国实验方剂学杂志

Objective:To investigate whether there is inflammatory reaction and cell pyroptosis in impaired glucose tolerance （IGT） rats induced by high-fat diet and the intervention effect of Huanglian Wendantang. Method:Healthy male SD rats were fed with 45% fat content diet for 20 weeks to replicate the IGT model. The rats in line with the model establish criteria were randomly divided into 3 groups， with 10 rats in each group， another 10 rats were selected as the blank control group. Huanglian Wendantang group was given 7.8 g·kg^-1·d^-1 compound decoction of Huanglian Wendantang， and the positive control group was given 0.05 g·kg^-1·d^-1 aqueous solution of metformin hydrochloride， with the dose volume of 10 mL·kg^-1·d^-1. The blank group and the model group were given the same volume of distilled water. After continuous intragastric administration for 4 weeks， the body weight， body length and abdominal circumferences were measured， the Lee's obesity index was calculated， and the levels of fasting plasma glucose （FPG） and 2-hour plasma glucose （2 h PG） were detected in each group. Serum interleukin-6 （IL-6） content was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of nuclear factor kappa B （NF-<italic>κ</italic>B）， cysteinyl aspartate specific proteinase-1 （Caspase-1） and NOD-like receptor protein 3 （NLRP3） in liver tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Immunofluorescence was used to detect the expression of Caspase-1 and NLRP3 in rat liver. Hematoxylin and eosin （HE） staining was used to observe the morphology of liver tissue. Result:Compared with blank group， the body weight， abdominal circumference， body length and Lee's index in model group were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， the levels of FPG and 2 h PG were significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， serum IL-6 content and NF-<italic>κ</italic>B， Caspase-1 and NLRP3 gene and protein expressions in liver tissue were significantly increased （<italic>P</italic><0.01）， immunofluorescence technique showed that Caspase-1 and NLRP3 expressions increased obviously in IGT rat model’s liver tissues， and inflammatory cells infiltration was observed obviously in HE stained rat liver tissue model. Compared with model group， Huanglian Wendantang and metformin hydrochloride groups could effectively reduce the body weight of IGT rats （<italic>P</italic><0.01） and abdominal obesity， and correct the levels of FPG and 2 h PG （<italic>P</italic><0.01）， while effectively reducing serum IL-6 content and gene and protein expressions of NF-<italic>κ</italic>B， Caspase-1 and NLRP3 in liver tissues of IGT rats， and alleviating inflammatory cell infiltration. Conclusion:Inflammatory response exists in IGT model rats induced by high-fat diet. Huanglian Wendantang can obviously reduce its inflammatory response and alleviate liver cell pyroptosis.

Mechanism of Huanglian Wendan Decoction in improving impaired glucose tolerance based on skeletal muscle NLRP3/caspase-1/IL-1β, IL-18 pathway / 中国中药杂志

This study investigated the mechanism of improving impaired glucose tolerance(IGT) of rats by Huanglian Wendan Decoction from the perspective of the skeletal muscle Nod-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/interleukin-1β(IL-1β), interleukin-18(IL-18) pathway. Healthy male SD rats were fed with the diet containing 45% fat for 20 weeks, accompanied by a high-temperature and high-humidity environment and an inactive lifestyle, for the establishment of the IGT rat model. The rats were divided into the blank control group, model control group, metformin hydrochloride group(positive drug group, 0.05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group(7.8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks, the obesity and glycemic indexes of all the rats were measured. The fasting serum insulin(FINS) level was determined by ELISA, with the insulin sensitivity index(ISI) and insulin resistance index(IRI) calculated. The mRNA and protein expression le-vels of nuclear factor kappaB(NF-κB), NLRP3, caspase-1, IL-1β and IL-18 in skeletal muscle tissue were detected by real-time polymerase chain reaction(PCR), Western blot and immunofluorescence. Compared with the blank control group, the model control group witnessed significantly increased mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18. As revealed by the comparison with the model control group, Huanglian Wendan Decoction could effectively regulate the obesity status, reduce body weight, correct the abnormal levels of 2-hour plasma glucose(2 hPG), insulin resistance index(IRI), insulin sensitivity index(ISI), and lower the mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1β and IL-18 in the skeletal muscle tissue of IGT rats. Combined with previous studies, the above results showed that the occurrence and development of IGT was closely related to inflammatory response and the classic pyroptosis pathway in skeletal muscle, such as NLRP3/caspase-1/IL-1β, IL-18. It is inferred that the mechanism of Huanglian Wendan Decoction was to alleviate insulin resistance(IR) and then reverse the course of IGT lies in the regulation of the abnormal insulin receptor signaling pathway based on the NLRP3 inflammasome pathway.

Effect of Qingre Jiedu Medicine in Treating Tumor and Regulating Signaling Pathway / 中国实验方剂学杂志

Tumor is a new organism caused by normal local tissue and cell proliferation under the effect of various tumorigenic factors. It is a serious threat to human health and life. In the theory of traditional Chinese medicine, heat poison that is similar to tumorigenic factor is one of the main causes of tumor. Modern studies have shown that Qingre Jiedu drugs have the effect in clearing heat, detoxifying, resisting bacteria and inflammation, improving immunity and resisting tumor. Therefore, Qingre Jiedu drugs are an important part of traditional Chinese prescriptions for the treatment of tumors. The research of Qingre Jiedu drugs has attracted more and more attention. Molecular pharmacological studies have shown that Qingre Jiedu drugs can play a significant role in the treatment of many kinds of tumors by regulating the targets in multiple cell signal transduction pathways. Based on literatures, due to the lack of specific review on heat and detoxification drugs for the treatment of tumor mechanism in recent years, Sophorae Flavescentis Radix, Scutellariae Radix, Hedyotis diffusa willd and Scutellariae Barbatae Herba with typical effect in clearing heat and detoxification were selected to study the tumor cell proliferation, invasion and metastasis and angiogenesis, and effects in reducing or eliminating chemotherapeutic drug resistance, inhibiting cell proliferation and inducing cell apoptosis of Sophorae Flavescentis Radix, Scutellariae Radix, Hedyotis diffusa willd and Scutellariae Barbatae Herba through Wnt/beta-catenin, phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt), Hedgehog signaling pathways. Apparently, heat and detoxification drugs can inhibit the occurrence and development of tumors as a result of the common action of various components in heat and detoxification drugs. Meanwhile, heat and detoxification drugs have the advantages over chemical therapy and surgical treatment. This provides a reference for the application and further study of heat and detoxification drugs in tumor treatment, and new theoretical support and action target of modern medicine for the mechanism of action of traditional Chinese medicine in treating cancer.

Intervention of Epimedium 95％ethanol elution section in potential biomarkers in urine of rats based on metabonomics / 中国药理学与毒理学杂志

OBJECTIVE To observe the effect of Epimedium 95％ethanol elution section (E95EE) on endogenous metabolism in the urine of normal rats using methods of metabonomics, and to study whether differential expressions of biomarkers can influence different systems of the body along with various body-fluids-cycle distribution. METHODS SD rats were randomly divided into blank control group and E95EE group(10 rats per group). The rats of E95EE group were ig administered with E95EE 17.1 g · L-1, once daily, for 20 d, while those of normal control group were ig given an equal volume of saline. On the day of the final E95EE administration, the urine of 12 h was collected for analysis by UPLC-TOF/MS. RESULTS This study identified nine differential endogenous metabolites (3-sn-phos-phatidate, 5, 10-methenyltetrahydrofolate, l-1-phosphatidylethanolamine, 1-acyl-glycerone 3-phosphate, N-formimidoyl-l-glutamate, keto-oxaloacetate, sulfurous acid, formyl-N-acetyl-5-methoxykynurenamin and N-acetyl-5-hydroxytryptamine) and six primary metabolic pathways [glycerophospholipid metabolism, vitamin A (retinol) metabolism, histidine degradation, tryptophan metabolism, acetyl-CoA biosynthesis from citrate, glycolysis and gluconeogenesis]. The possible role of protection of E95EE discovered in the nervous and cardiovascular systems was displayed by the decreased level of 3-sn-phosphatidate and an increased level of N-formimidoyl-l-glutamate. However, the possible toxicity of E95EE on neoplastic prevention was achieved by reducing the level of l-1-phosphatidylethanolamine and 5,10-methenyl tetra-hydrofolate. CONCLUSION E95EE can produce both protection and toxicity on nervous, cardiovascular and immune systems, as well as on tumor-associated diseases. The mechanisms may be related to metabolic pathways of triglycerides, vitamin A, tryptophane and histidines.

Comparative study on serum pharmacochemistry of Gardenia jasminoides under physiological and pathological status / 中草药

Objective: By means of serum pharmacochemistry combined with modern chromatographic technology to elucidate the material basis for efficacy of aqueous extract from Gardenia jasminoides (AEGJ), which has antipyretic activities with bitter-cold property. Methods: Waters 2695 HPLC was employed as the detection system, based on the same chromatographic eluted condition, physiological and DNP-induced fever syndrome serum samples, collected from hepatic portal vein of rats after ig administration of AEGJ, were under determining to compare the absorptive variation of the chemical compounds. Results: The chromatographic fingerprints of common peaks for six batches of AEGJ were established to ascertain the optimized administration dosage, blood collection time, and the method used to process the serum. Compared with the normal administrated group under physiological status of rats, we found that the significant increment absorption of jasminoidin and the generation of two new migration compounds in the DNP fever syndrom model group. Conclusion: The increment absorption and the migration of chemical compounds in serum of DNP fever syndrome pathological model of rats might reasonably lead us to consider these chemical compounds as symbols of the bitter-cold property of G jasminoides.

A comparative study on alkaline ashing method and chloric acid digestion method for determination of human milk iodine / 中国地方病学杂志

Objective Take alkaline ashing method as golden standard to explore the accuracy of chloric acid digestion method in determination of human milk iodine. Methods Sixty one breast milk samples collected in Hexi district of Tianjin was measured by the method for determination of iodine in foodstuff by As3+-Ce4+ catalytic spectrophotometry (referred to as the alkaline ashing method) published in 2008 and the method for determination of iodine in urine by As3+-Ce4+ catalytic spectrophotometry(referred to as acid digestion) published in 1999, respectively. were highly correlated(r = 0.960, t = 26.3, P < 0.01), and the regression equation was (Y) = - 28.1 + 0.808X, in which X was independent variable, that is the results of alkaline ashing method; (Y) was dependent variable, that is the estimated data of chloric acid digestion method. The average difference of the results measured by the two methods was 68.3 μg/L, and the results from chloric acid digestion was 38.9% which lower than that of alkaline samples were diluted by 3,4 and 5-fold and then digested by chloric acid, the liquid clarification rates were 80.3% ashing and chloric acid digestion method were, respectively, 165.4, 110.0 μg/L. Conclusions Compared with alkaline ashing method, the results determined by chloric acid digestion method are significantly lower. It is suggested that there are systemic errors in chloric acid digestion method, which means that alkaline ashing method can not be replaced by the chloric acid digestion method.

The effect of insulin-like growth factor-Ⅰ on thyroid morphologic change of mice with iodine deficiency and iodine excess / 中国地方病学杂志

Objective To study the expression level of thyroid insulin-like growth factor-Ⅰ (IGF-Ⅰ) in iodine deficiency and excess mice and the effect of thyroid gland IGF-Ⅰ in the thyroid morphological change. Methods Forty-eight Balb/c mice were chosen as studied objects,weighing about 16 g. They were divided into three groups: low iodine(LI,iodine content of 50 μg/kg in feed,drinking deironized water) group,normoi(NI,iodine content of 300 μg/kg in feed,drinking deironized water) group and high(HI,iodine content of 300 μg/kg in feed,iodoine of content 14 700 μg/kg in drinking) group,16 mice in each group. Mice were put to death after 12 weeks and taken out of their thyroid gland. The body weight,absolute and relative weights of thyroid gland were measured and the morphological change of thyroid gland were observed under microscope. The expression levels of thyroid gland IGF-Ⅰ mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively. Results There were statistical significances between groups of thyroid absolute and relative weights(F = 315.881,405.921,all P < 0.01). LI group [(10.71±4.03) mg,(44.98±15.39)mg/100 g body weight]and HI group [(3.42±1.17)mg,(13.50± 3.89)mg/100 g body weight]had heavier thyroid absolute and relative weights than NI group[(2.11±0.53)mg,(8.35±1.98)mg/100 g body weight,all P < 0.01]. Under microscopy,the thyroid follicle capacity grew down and the follicle quantity grew up in LI group,the epithelium was stylolitic,the colloid diminished or absence in follicular cavity,while HI group presented colloid accumulation without follicular hyperplasia. The expression level of thyroid gland IGF-Ⅰ mRNA in LI group(1.03±0.32) was more than that in NI(0.65±0.19) and HI(0.59± 0.20) groups(F= 7.518,P< 0.01). In contrast to NI,there was no difference in the expression level of thyroid gland IGF-Ⅰ mRNA in HI group(P > 0.05). The brownish particles of LI group were more than NI and HI groups in the thyroid follicle epithelium by immunohistochemistry,while HI group was less than NI group. Conclusions The mice of iodine deficiency presented follicular hyperplasia goiter,the mice of iodine excess presented colloid accumulative goiter. The change of IGF-Ⅰ mRNA and protein expression may participate morphologleal change,indicating autocrine IGF-Ⅰ of thyroid gland may play an important role in regulating goiter formation.

Serum hyaluronan levels in oral cancer patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Hyaluronan (HA) is most likely associated with tumor invasion and metastasis. Studies have shown that HA levels are often increased in serum of patients with various malignant tumors. The purpose of this study was to determine the levels of serum hyaluronan in patients with oral cancer and evaluate the value of serum HA in adjuvant diagnosis, staging and monitoring treatment response in these patients.</p><p><b>METHODS</b>Eighty-four hospitalized patients with oral cancer, 65 patients with benign tumors in the oral and maxillofacial region and 67 healthy individuals were included in this investigation. Venous blood was collected from these patients and the healthy individuals before therapy. One week after therapy, venous blood was collected once again in 43 patients with oral cancer. Serum samples were obtained and serum HA levels examined.</p><p><b>RESULTS</b>The serum HA concentration was significantly higher in oral cancer patients than in patients with benign tumors and in healthy controls (P<0.05). The serum HA level in patients with stages III and IV disease was higher than in patients with stages I and II disease, but there was no significant difference in the HA level between stages I and II nor between stages III and IV (P>0.05). After a complete treatment the HA levels in patients with oral cancer became lower than before treatment, but the difference was not significant (P>0.05).</p><p><b>CONCLUSIONS</b>The results of this study suggest that the determination of HA levels may provide additional information in diagnosis of oral cancer, but its usefulness as an adjunct in clinical staging and in monitoring treatment response was limited.</p>

Effect of excessive iodine intake on sodium-iodide symporter mRNA and protein expression of breast in lactating rats / 中国地方病学杂志

Objective To study effect of excessive iodine intake on sodium-iodide symporter(NIS)mRNA and protein expression of breast in lactating rats.Methods60 Wistar rats,having been weaned for one month,were randomly divided into three groups according to their body weights,I.e,①normal iodine(NI,30 rats);②ten fold high iodine(10 HI,15 rats);③one hundred fold high iodine(100 HI,15 rats).Eating food containing iodine of 300μg/L and drinking water of iodine at 5,1845,20 295μg/L,respectively.After fed for 3 months,the rats mated and had offspring,and urine and milk iodine of lactating rats were determined by As-Ce-catalytic spectrophotometric method.Their marmnary glands were sampled at lactation day 10.Then NIS mRNA expression by RT-PCR was determined and NIS protein by immunohistochemistry(SABC)was observed.Results The urine iodine of 10 HI group(3597.5μg/L)and 100HI group(25 404.3μg/L)increased obviously compared with that of NI group(344.7μg/L).The milk iodine of 10HI group(27.1×103μg/L)and 100HI group(191.0×1μg/L)was higher than that of NI group(6.0×103μg/L),but the increased fold of milk iodine was not paralleled with that of urine iodine.Difference of NIS mRNA expression was significant(F=24.19,P＜0.01)among the groups,and the NIS mRNA expression in 10HI(1.250±0.034)and 100HI(1.272±0.039)group were less than that in NI (1.532±0.044)group(P＜0.01).The breast NIS mRNA expression in lactating rats(1.532±0.044)was significantly higher than that in unlactating rats(0.879±0.018,P＜0.01).With the increasing iodine uptake,NIS protein expression decreased.Conclusions The NIS mRNA and protein in rat breasts is down-regulated by excessive iodine intake.So increasing extent of milk iodine concentration is inhibited,which is important to prevent off-spring from getting excessive iodine intake from parental generation.

Fibular flap combined with lateral crural flap for reconstruction of oral and maxillofacial defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effect of fibular flap combined with lateral crural flap for reconstruction of oral and maxillofacial defect.</p><p><b>METHODS</b>Based on the peroneal vascular system, fibular flap combined with lateral crural flap were used for reconstruction of oral and maxillofacial defect. The fibular flap was used for bone defect, and the lateral crural flap was for the reconstruction of soft tissue defect of the oral cavity and pharynx.</p><p><b>RESULTS</b>During the period of Mar. 2005 to Mar. 2007, 26 cases were treated, including 25 cases of defects after tumor resection and 1 case of bilateral maxillary defect. The flaps were harvested without any injury to the peroneal vascular system and perforator. All the flaps were survived. One case of arterial insufficiency and one case of venous thrombosis occurred 12 hours and 24 hours after operation, but the flaps were salvaged after urgent re-operation. All the patients were followed up for 6 months to 2 years. The patients acquired satisfactory appearance with normal social life.</p><p><b>CONCLUSIONS</b>For complicated oral and maxillofacial reconstruction, fibular flap combined with lateral crural flap can achieve good reconstruction results and could be selected as the first line treatment.</p>

Glandular odontogenic cyst: report of two cases with cytokeratin 18 expression / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To report two cases of glandular odontogenic cyst and examine its cytokeratin 18,19 expression.</p><p><b>METHODS</b>Two cases of glandular odontogenic cyst were reported and studied. The cytokeratin 18, 19 expression in these two cases were also investigated using immuno-histochemical staining as well as in the situ hybridization of the cyst epithelium.</p><p><b>RESULTS</b>Histo-pathological examination revealed that ciliated columnar cells, squamous cells and low-columnar cells were found in the superficial layer of the lining epithelium. Several minor salivary glands, mainly composed of seromucous cells were observed near the satellite cyst. CK18 were expressed in all layers of the lining epithelium of varying intensity. CK18 was negative in lining epithelium of the daughter cyst, but CK19 was positive. CK18-mRNA was expressed in all the layers of the lining epithelium, the salivary glands and daughter cysts.</p><p><b>CONCLUSIONS</b>Histological features and CK18 expression may be indicative of the possibility of salivary glandular and odontogenic differentiation.</p>

Simultaneous screening program for newborns hearing and ocular diseases / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To explore the model and the feasibility of newborn hearing and ocular disease simultaneous screening program and to study the birth prevalence of newborn hearing loss and newborn ocular diseases.</p><p><b>METHODS</b>The universal newborn hearing screening (UNHS) was performed using transient otoacoustic emission (TEOAE) in well baby nursery and by a two-stage TEOAE and auto auditory brainstem response (AABR) protocol in neonatal intensive care unit (NICU). The UNHS was simultaneous done with newborn ocular disease screening program. The examination technical method was following: the response to light, external inspection of the eyes and lids, pupil examination, red reflex examination, funduscope examination after pupil dilation for referral (for all newborn in NICU). The infants who were referred by two-stage hearing screening and/or had high-risk factors of hearing loss received following-up and routine audiological evaluation and personalized intervention from 6 months to 3 years of age. The cases had positive sign and (or) abnormal results of the ocular disease screening were referred for further examination by pediatric ophthalmologists.</p><p><b>RESULTS</b>A total of 16 800 children born in Jinan Maternal and Child Hospital from October 1, 2002 to April 30, 2005. Of these infants, 15 398 cases (91.7%) had access to the simultaneous screening program for hearing and ocular diseases. The incidence of congenital sensorineural hearing loss (SNHL) among infants who did UNHS was 0.312% (48/15 398) in bilateral and 0.227% (35/15 398) in unilateral; Of the 4 cases of congenital SNHL complicated with newborn ocular diseases: 1 profound SNHL (bilateral), auditory neuropathy with congenital cataract (bilateral), 1 mild SNHL (bilateral) with membrana papillaris perseverance (left) and 1 mild SNHL (bilateral) with retina vein dilatation (bilateral), 1 mild SNHL (right) with persistent hyaloid artery (bilateral). In all 15 398 newborns, 15 neonates with congenital cataract were detected (22 eyes, 0.10%). Twenty seven neonates with less than 1500 g birth weight admitted to NICU, retinopathy of prematurity was detected in 3 neonates (6 eyes).</p><p><b>CONCLUSION</b>Hearing loss and ocular diseases was not rare in neonatal and infancy. Newborn hearing and ocular disease simultaneous screening program was not only feasible but also effective in detecting hearing loss and (or) ocular disorders. Early intervention was important for the prevention or treatment of neonatal hearing loss and (or) ocular diseases, such as newborn hearing loss with congenital cataract, retinopathy of prematurity and so on.</p>

Cytokeratin 18 and their gene expression in jaw odontogenic keratocyst epithelial lining / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To examine cytokeratin 18(CK18) and it's gene in jaw odontogenic keratocyst (OKC) epithelial lining.</p><p><b>METHODS</b>The epithelial linings of 32 cases were subject to monoclonal antibody immunohistochemical staining for CK18, CK8 and CK19. RT-PCR and in situ hybridization for CK18 mRNA were conducted in 12 of 32 cases in keratocyst epithelial cell linings.</p><p><b>RESULTS</b>In 17 cases, CK18 were observed in keratinized surface layers, though weakly positive. In 27 cases, CK18 were positive in the granular cell layers. CK18 were also positive in the spinous cell layers in 14 cases. In all cases, CK18 was negative in basal cell layers. By RT-PCR, 4 cases expressed CK18 strongly, 8 cases weakly. By in situ hybridization, 8 cases expressed CK18 mRNA positively in both spinous and granular cell layers, and 4 cases positively in basal and keratinized cell layers. CK8 were expressed in basal cell layers of keratocyst epithelial linings. In 23 cases, CK19 were expressed in surface cell layers of keratocyst epithelial linings.</p><p><b>CONCLUSION</b>The expression of CK18 in keratocyst epithelial linings transfers from basal cell layer to spinous layer. The expression of CK18 immunohistochemical staining and CK18 mRNA in situ hybridization are different, which shows CK18 might be related to proliferation of OKC epithelial linings. That suggests the existence of regulation of CK18 and CK18 mRNA expression.</p>

Comprehensive protocol for diagnosis and treatment of childhood neuroblastoma--results of 45 cases / 中华儿科杂志

<p><b>OBJECTIVE</b>The aim of the paper was to improve the prognosis of neuroblastoma (NB) stage III and IV in children through the comprehensive therapy including chemotherapy, delayed tumor resection, autologous stem cell transplantation (ASCT) and inducing differentiation and to analyze the factors affecting the prognosis.</p><p><b>METHODS</b>Newly diagnosed neuroblastoma patients seen from Oct.1998 to Dec.2003 were divided into high, medium and low risk groups depending on clinical stage and age. Comprehensive protocol included accurate staging, delayed and/or second tumor resection for stage III and IV patients, chemotherapy of different intensity mainly composed of cell cycle nonspecific drugs and 13-cis-retinoid for inducing cell differentiation. ASCT was given at the end of therapy for high risk group.</p><p><b>RESULT</b>Forty-five patients, 6 months to 11 years of age, 32 males and 13 females, were analyzed. Of them, 15 were found to have the tumor in adrenal gland, 12 had the tumor extended to the retro-peritoneal space, while in 15 cases the tumor was beside the spinal column in chest and in 3 the tumor was located in other places. Nine cases had stage I, 1 case had stage II, 8 cases had III, 26 cases had stage IV and 1 case had stage IVs of the tumor. Depending on the age and stage of the tumor, 26 cases were aligned into high risk protocol, 10 into medium risk and 9 into low risk groups. Thirty nine cases were treated as planned. Eleven of them received ASCT including 2 cases who received second ASCT. Of the thirty-nine patients, 31 achieved complete remission (CR) and 8 partial remission (PR) after surgery and/or chemotherapy. During up to 21 months median following up period (range 14 to 64 months), 24 cases (62%) kept CR (median 22 months) and 4 survived with stable disease. The survival rate (SR) was 72%. Eleven cases died of relapse and disease progression. No death occurred from treatment complication. Statistical analysis showed that the age older than 18 months, and stage III and IV of the tumor were the factors predicting poor prognosis (P = 0.04 and 0.003, respectively). Patients who had the tumor originated from the retroperitoneal space, who had incomplete tumor resection, and those who did not receive ASCT had poorer prognosis, but the differences were not significant (P = 0.092, 0.55 and 0.60, respectively).</p><p><b>CONCLUSION</b>The comprehensive protocol seemed to be reasonable. Age older than 18 months, and stage III and IV were the factors suggesting poor prognosis. The origin of the tumor, completeness of tumor resection, and use of ASCT had no significant impact on the prognosis.</p>

Cytokeratin18, 13 and their gene expression in post-operative maxillary cyst linings with metaplastic epithelium / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium.</p><p><b>METHODS</b>CK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases).</p><p><b>RESULTS</b>The expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA.</p><p><b>CONCLUSIONS</b>These results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.</p>

Examination of serum hyaluronic acid level in patients with oral and maxillofacial malignancy / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To determine the level of hyaluronic acid (HA) in serum of patients with oral and maxillofacial malignancy and to investigate the correlation between the levels of serum HA and stage of the malignant lesions and treatment response.</p><p><b>METHODS</b>44 patients with oral and maxillofacial malignancy were analyzed and 24 healthy individuals served as controls. Venous blood was collected from the patients before treatment and the healthy individuals. One week after therapy, venous blood were collected in 24 patients once again. Serum levels of HA were measured with quantitative radioimmunoassay (RIA).</p><p><b>RESULTS</b>Before treatment, the serum HA concentration in patients with oral and maxillofacial malignancy was significantly higher than that of the controls (P < 0.05). Also, the serum HA concentration in patients with OSCC was significantly higher than that of the controls (P < 0.05). No difference was noted in serum HA concentration between patients with salivary ACC and the control group (P > 0.05). The serum HA concentration of patients in stage III and IV was significantly higher than that of patients in stage I and II (P < 0.05). Serum HA levels decreased in patients after a complete treatment, but the difference was not significant (P > 0.05).</p><p><b>CONCLUSION</b>Serum levels of HA may be useful in diagnosis of OSCC and was associated with clinical stages in patients with oral and maxillofacial malignancy. However, it may not be contributory to monitoring treatment response in patients with oral and maxillofacial malignancy.</p>