Objective To compare the purification process of two types of ceramic hydroxyapatite(CHT I and CHT II)to remove the aggregates from two monoclonal antibodies(mAb 1 and mAb 2).Methods All the chromatography runs were performed on AKTA AVANT 150 with Tricon 10/50 column.The dynamic binding capacity( DBC) of two types of CHT was studied firstly, and then purification research was carried out selecting the suitable DBC.The column was equilibrated with 5 mmol/L sodium dihydrogen phosphate pH 6.5, and then was eluted with gradient buffers which were 10 mmol/L sodium dihydrogen phosphate pH 6.5 and 2 mol/L sodium chloride pH 6.5.Aggregate content in loading and elution pool was evaluated by size exclusion chromatography.Scale-up process was carried on 20 cm height chromatography column XK16/40.Results DBC of CHT I for mAb 1 was 40 mg/mL and mAb 2 was 45 mg/mL.After purity, monomer content of mAb 1 reached 98.6% and yield was 92.5% and monomer content of mAb 2 reached 98.8%and yield was 91.5%.DBC of CHT II for mAb1 was 16 mg/mL and mAb 2 was 20 mg/mL.After purity, monomer content of mAb 1 reached 99.8% and yield was 91.8% and monomer content of mAb 2 reached 99.9% and yield was 92.2%.Conclusion Two types of CHT both can remove aggregates effectively from monoclonal antibodies when aggregate content reaches more than 10%, and results conform to the regulations.CHT I has higher dynamic binding capacity than CHT II, and CHT II is superior to CHT I in removing aggregate efficiency.The purification process is simple and can be easily scaled up in pilot and manufacture.Therefore, it meets the requirement pilot and scale production.

Objective To analyze the infection status and variation tendency of chlamydia trachomatis (CT) ,Neisseria gonorrhoe-ae(NG) and ureaplasma urealyticum(UU) in female genital tract in northeast Sichuan province during 2005 -2012 to provide the laboratory basis for their diagnosis and treatment Methods The real-time fluorescent quantitative PCR (RT-PCR) was applied to detect the CT DNA ,NG DNA and UU DNA in 1 386 samples from female genital tract and the detection results were performed the statistical analysis .Results The total positive rate of these 3 kinds of pathogens was 62 .8% (871/1 386) .Among the simple in-fection ,UU had the highest positive rate(48 .0% ,665/1 386);the positive rates of CT and NG were only 2 .2% .In the mixed infec-tion ,the positive rate of CT + UU was highest(6 .5% ,90/1 386) ,while which of UU + NG ,CT + NG and CT+ NG+ UU was 2 .5% (35/1 386) ,0 .4% (5/1 386) and 1 .1% (15/1 386) respectively .In different age groups ,the positive rate in the age <20 years old group was 49 .3% ,while which in the age >20 years old groups were all more than 60% .The positive rate of the CT ,NG and UU pathogens in females was in continuous high level during 2005 -2012 ,and which totally showed an increasing tendency . Conclusion CT and UU are the main pathogens in female genital tract infection in this region ,and the positive rate of genital tract infection in females aged more than 20 years is higher ,the infection rate of these 3 kinds of pathogens demonstrates the increasing trend year by year ,so more attention should be paid to the detection of CT and UU in this group for guiding the clinicians to con-duct the diagnosis and treatment .

Objective To study the correlation between pain, lower limb strength and physical function in patients with primary osteoporosis. Methods Fifty-seven female patients diagnosed with menopause-related low bone mass or primary osteoporosis using a GE calcaneus bone density detector were involved in this study. The muscle strength of their lower limbs was tested with a Biodex system 4 machine. Pain was assessed with a visual analogue scale, and physical function (PF) with the SF-36 instrument. Results Low back pain was significantly correlated with PF, and so was leg pain. Leg pain was also significantly correlated with the strength of the extensors of the dominant leg during low velocity and medium velocity movement. Leg pain was not, however, significantly correlated with the strength of the flexors of the dominant leg during low and medium velocity movement. Conclusions Pain predicts poor physical function in patients with primary osteoporosis or low bone density. During low and medium velocity movement, leg pain significantly predicts poor muscle strength in the extensors of the dominant lower leg, but it has no correlation with the muscle strength of the flexors.

Objective To investigate the characteristic,diagnosis,clinical staging, treatment and clinical prognosis of occult breast carcinoma (OBC). Method Forty-six cases of OBC were analyzed retrospectively with the clinical and follow-up information that were confirmed by postoperative pathologic diagnosis from November 1981 to November 2005. Results All patients showed axillary node enlargement as the first sign and were operated.The operation included axillary node excision in 2 patients,radical mastec-tomy or modified radical mastectomy in 44 patients. Forty-five cases got follow-up for 1-22 years,33 cases had existed 3 years,18 cases had existed 5 years,8 cases had existed 10 years. Conclusions For axillary mass which causes are uncertain ,the possibility of OBC should be considered .Meanwhile excision and pathological examination is necessary.The metastatic histological structure and immunohistochemical index of the axillary nodes usually provide important clue for the source of this tumor.Radical or modified mastectomy is the best method, and pest-operative chemotherapy and/or radiotherapy should be done. It has been showed that targeted therapy is very important to breast cancer with C-erbB-2 positive recently.To the cases that neck lymphatic metastasis is M4G3 positive by immunohistochemical examination and no primary focus clinically, the diagnosis of OBC should be considered. The cases without primary focus have better prognosis than those with primary focus.

OBJECTIVETo study the correlation between arachidonic acid (AA) and acute red blood cells damage in rats, and to build a model with hidden blood loss in vivo, and to explore the pathological mechenism of hidden blood loss.

METHODSA total of 50 male adult Sprague-Dawley rats weighing (200 ± 20) g were randomly divided into five groups (n = 10): control group and four experimental groups. The rats in the experimental groups were given 0.5 ml different concentrations of AA dilu- ents, 5, 10, 20, 40 mmol/L respectively. The blood samples were collected from orbital venous at the beginning and 24, 48, 72 hours after administration. Then the changes of hemoglobin (Hb) ,red blood cell count (RBC), glutathione peroxidase (GSH- PX) activity, total superoxide dismutase (T-SOD) activity and hydrogen peroxide (H202) in the blood samples were tested.

RESULTSSignificant hidden blood loss occurred when the concentration was 10 mmol/L in the experimental group, with the RBC and Hb sharply reduced in blood samples. The Hb and RBC were reduced in all the experimental groups and control group at 24 hours after administration, while in the experimental groups they changed more obviously. The GSH-PX activity, T-SOD activity and H₂O₂were also significantly reduced in all groups, and the changes showed significant differences. The Hb and RBC were relatively stable in the control group and the experimental groups at 48 hours after administration; while GSH-PX activity, T-SOD activity and H₂O₂were all significantly decreased, and the changes in the experimental groups were more notable.

CONCLUSIONElevated levels of AA in the blood causes oxidative stress in the red blood cells, leading to the damage of red blood cells and hemoglobin, which is responsible for hidden blood loss.

Animals ; Arachidonic Acid ; toxicity ; Erythrocytes ; drug effects ; metabolism ; Glutathione Peroxidase ; blood ; Hemoglobins ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood

Objective To study the clinical features of congenital glucose-galactose malabsorption (CGGM),and to improve the understanding of CGGM.Method Clinical manifestations and treatment process of one patient with CGGM in our hospital were retrospectively analyzed.From 1966 to 2016 May,Chinese medical database and PUBMED were searched using Malabsorption syndrome,dehydration,hypernatremia , diarrhea , newborn , carbohydrate metabolism ,andglucose/galactose malabsorption as key words.The clinical features of CGGM reported in literatures were summarized.Result The patient in our hospital was a full-term female infant naturally delivered.The onset of the disease was on the 9th day after birth,and the clinical manifestations included severe diarrhea,severe dehydration,hypernatremia,metabolic acidosis and malnutrition.After intravenous infusion and symptomatic treatment,dehydration,hypernatremia and metabolic acidosis were corrected.However,there was no improvement of diarrhea characterized with watery and acidic stools,and neither was weight gain.Glucose loading test was negative,and fructose loading test was positive.Diarrhea was improved markedly using diagnostic carbohydrate-free formula,so CGGM was diagnosed clinically.SLC5A1 homozygous IVS7-2 A ＞ G mutation was detected which confirmed the diagnosis of CGGM.With carbohydrate-free formula feeding,the body weight of the infant was increased.Followed up for 2 months now,her body length and body weight were at P25 and P22 on growth curve respectively,and no obvious neurological sequela was observed.Our literature review revealed 7 reports including 48 cases of CGGM children.Literature review showed that:most children with CGGM (79.2％) had the onset within 7 days of life;main clinical features included diarrhea (100％),dehydration (100％),and malnutrition (54.2％);22.9％ of patients with carbohydrate-free formula and 27.1％ with fructose matrix formula were fed well;no death was detected,77.1％ had normal weight gain,and 91.7％ had normal development of the nervous system.Conclusion CGGM is rare.The symptoms include severe watery and acidic stools with onset during neonatal period.CGGM is associated with severe complications such as hypertonic dehydration and hypernatremia.The diagnosis is established based upon typical clinical manifestations,sugar loading test and SLC5A1 gene detection.Carbohydrate-free formula feeding is effective.

[Summary] A case of primary hyperparathyroidism ( PHPT ) complicated with Graves′disease was reported.The parathyroid lesion( s) could not be identified by repeated MIBI and ultrasonography tests.With the control of hyperthyroidism, medical therapies of hypercalcemia were still not effective, the serum calcium levels continued to be high.Thus, the decision to operate was made.The pathological findings confirmed the diagnosis of parathyroid adenoma.For PHPT patients with clear surgical indications, even though the pre-operative localizing tests are negative, operation is still worth to try.

Objective To investigate the clinical characteristics of neonatal multi-system Langerhans cell histiocytosis.Methods A female neonate with multi-system Langerhans cell histiocytosis in Guangzhou Women and Children's Medical Center in March,2013 was reported and related literature was reviewed.The clinical manifestations,and laboratory test,imaging examination and biopsy results were analyzed.Results The neonate had onset of jaundice and rash,then was found to have hepatomegaly,splenomegaly,lymphadenopathy and abnormal blood test.The lymph node biopsy revealed infiltration of Langerhans cells and eosinophils,which were positive for CDla/S-100 stains.She was diagnosed with multi-system Langerhans cell histiocytosis.Sixty-two cases of neonatal multi-system Langerhans cell histiocytosis were reviewed in the literature,and the most common clinical manifestations included rash,hepatomegaly,respiratory symptoms,imaging changes,abnormal blood test,digestive symptoms and lymphadenopathy.The most frequently involved organs included lung,liver,spleen,bone marrow,bone,gastrointestinal tract,lymph node and mucosa,among which the risky organs (liver,spleen,blood system and lung) accounted for 88.7％(55/62).Conclusions The incidence rate of multi-system Langerhans cell histiocytosis is low in neonates,and it is characterized by multisystem involvement,especially the liver,spleen,blood system,lung and skin.For the neonates diagnosed without intrauterine infections,besides skin biopsy,lymphadenopathy should be dynamically observed and lymph node biopsy should be done earlier so as to establish an early diagnosis.

Objective To study the therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells (hAMSCs) in rats with experimental type 1 diabetes mellitus (T1DM).Methods The hAMSCs from human amnion were isolated and cultured in vitro,then phenotype was analyzed by flow cytometry.T1DM were produced by administering streptozocin to rats.The rats were divided into normal control group (n =6),T1 DM model group (n =6),medium treated group (n =6),hAMSCs transplanted group(n =6),and insulin treated group(n =6).5 × 106of hAMSCs or vehicle were administered to rats via sublingual vein.Blood glucose levels of rats were recorded weekly in the groups for six weeks by Blood Sugar Meter.At the end of 6 weeks after hAMSCs transplantation,concentrations of plasma insulin were detected by ELISA; histopathological changes of pancreas,surviwl and differentiation of transplanted hAMSCs in pancreatic tissue were studied with HE staining,and immunofluorescence staining; percentages of lymphocyte subsets in peripheral blood mononuclear cells were determined by flow cytometry ; concentrations of plasma cytokines were determined by cytometric bead array.Results After hAMSCs transplantation,blood glucose levels in rats with T1DM were decreased (P ＜ 0.01),while concentrations of plasma insulin were increased significantly (P＜0.01).At 6 weeks,cell-treated animals showed an improvement in pancreas damage ; the percentages of CD4 + IFN-γ+ (Th 1) and C D4 + interleukin (IL)-17 + (Th 17) cells were reduced (all P＜0.05),while the percentages of FoxP3-positive regulatory T cells (FoxP3 +Treg) and CD4+ IL-4+(Th2) cells were increased (all P＜0.01) ; plasma concentrations of interferon-γ,IL-2,and tumor necrosis factor-αwere decreased (all P＜0.01),but IL-4 level was increased (P＜0.05).No histological evidence of insulin producing cells from hAMSCs was seen within pancreas.Conclusions hAMSCs may reduce blood glucose and alleviate the islet damage in rats with T1 DM,which is related to their potential to up-regulate FoxP3 +Treg cells.

Objective To study the effects of intermittent low?dose administration of recombinant human parathyroid hor?mone (1-34) [rhPTH(1-34)] in the expression of Osterix (Osx) during early stage of fracture healing. Methods Forty?eight 2?month old male Sprague?Dawley rats were underwent close unilateral femoral fracture and intramedullary nail fixation. The sub?jects were divided into 2 equal groups randomly:treatment group undergoing subcutaneous injection of rhPTH(1-34) 10 mg/kg/d immediately after the operation and control group undergoing subcutaneous injection of normal saline of the same dose. Six rats in each group were sacrifice at 2, 7, 14, and 21 days after operation. X?ray photography study was conducted at 7, 14 and 21 days. Tissue RNA and protein were extracted from the bone tissues of bilateral femurs and the expression levels of Osx mRNA and pro?tein were evaluated via real time quantitative PCR and Western?blotting. Results Fracture healing was significant at 14 days af?ter operation, and the progress of fracture healing was better in the rhPTH(1-34) group than in the control group at 14 and 21 days. The relative expression of Osx mRNA and protein in the fractured femurs of the rhPTH(1-34) group (5.02±0.5 and 10.03±0.8 for Osx mRNA, 3164.03±131 and 3509.02±126 for protein) at 14 and 21 days after the operation were significantly higher than those of the control group (2.30±0.4 and 4.01±0.7 for Osx mRNA, 1053.04±121 and 2721.03±123 for protein). However, there was no significant difference at 2 and 7 days after operation between the rhPTH(1-34) and control group. Conclusion Intermittent low?dose administration of rhPTH(1-34) up?regulates the expression levels of osteogenesis?specific Osx mRNA and protein in rats. It will accelerate the early phase of fracture healing process.