Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

Objective To discuss the valuable influence of the total-body massage on the facilitation of the rehabilitation for the women in puerperium.Methods Both Chinese traditional medical total-body massage and the biological information theories and its methodology were employed in dealing with 50 cases of puerperants without contraindications,who had labored naturally 12 hours before and who had received cesarean sections 48 hours before. The results were compared with the control group in which conventional preventive health care for puerperants was adopted.Results Notable differences exist in the aspect of lochia,defecation,lactation and emotions between the two groups.Conclusion Significant benefits can be obtained from the massage to facilitate the rehabilitation of the women in puerperium while promoting their quality of lives and broadening the coverage of services of the preventive health care for the puerperants.

OBJECTIVETo compare the therapeutic effects of peripheral facial paralysis in acute stage by different interventions and explore the better treatments of peripheral facial paralysis.

METHODSOne hundred and thirty one cases of Bell's facial paralysis were randomly divided into three groups. In acupuncture group (44 cases), Dicang (ST 4), Jiache (ST 6), Hegu (LI 4), Yangbai (GB 14) and Taiyang (EX-HN 5), etc. were applied; in electroacupuncture group (45 cases), the selection of acupoints and needling method were same as those in acupuncture group, and the electroacupuncture therapy was applied on Dicang (ST 4), Xiaguan (ST 7), Yangbai (GB 14) and Taiyang (EX-HN 5) in acute stage; in medication and acupuncture group (42 cases), Prednisone and Acyclovir were taken by oral administration, Vitamin B1 and Vitamin B12, were applied by intramuscular injection in acute stage, and acupuncture was applied by the way which was same as that in acupuncture group during quiescent and recovery stages. The curative effects were evaluated by House-Brackmann Grading Scale, and the failed rates were observed by follow-up after one and three months.

RESULTSThe cured and markedly effective rates were 79.6% (35/44), 93.4% (42/45) and 78.6% (33/42) respectively in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.05). The cured rates above tympanichord were 54.2% (13/24), 85.2% (23/27) and 48.0% (12/25) in acupuncture group, electroacupuncture group and medication and acupuncture group, and the result in electroacupuncture group was superior to those in acupuncture group and medication and acupuncture group (P < 0.01). There was no significant differences of cured rates below tympanichord among three groups (P > 0.05); and the failed rate in electroacupuncture group was much lower than those in acupuncture group and medication and acupuncture group by follow-up after one and three months (all P < 0.01).

CONCLUSIONThe peripheral facial paralysis is effectively treated by electroacupuncture in acute stage, and it suggests that electroacupuncture should be applied early during the acupuncture treatment of peripheral facial paralysis.

Acupuncture Points ; Acupuncture Therapy ; Acyclovir ; administration & dosage ; Adolescent ; Adult ; Aged ; Electroacupuncture ; Facial Paralysis ; drug therapy ; pathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Prednisone ; administration & dosage ; Treatment Outcome ; Young Adult

BACKGROUND: The effect of pituitary adenylate cyclase activating polypeptide (PACAP) during traumatic brain injury (TBI) and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI. METHODS: Male Sprague Dawley rats were randomly divided into 3 treatment groups (n=6, each): sham-operated, vehicle (normal saline)+TBI, and PACAP+TBI. Normal saline or PACAP (1g/5L) was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4+ and CD8+ T cells in blood and the spleen were detected via flow cytometry. RESULTS: In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4+ CD8– lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI+vehicle, and CD4– CD8+ were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4+, and decreased CD8+, T lymphocytes in blood and the spleen. CONCLUSION: Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

Objective To evaluate the clinical diagnostic value of nucleic acid amplification (TB- RNA),bacteriophage-based assay,3D culture and smear on the detection of Mycobacteria tuberculosis.Methods 291 clinical sample including 110 sputum,54 thoracic fluid,37 throat swab,31 bronchial fluid,13 cerebrospinal fluid,12 urine,8 lymph fluid and 20 others (pericardial effusion,feces, blood and abdominal fluid) and gynecological specimen (including 6 leucorrhoea and menstrual blood) were analyzed by these four methods.Results Among the 291 clinical samples,the positive rate of mycobacteria tuberculosis for TB-RNA,bacteriophage-based assay,3D culture and smear were 37.1%,28.9%,27.5% and 10.3%.The sensitivity and specificity of the TB-RNA,bacteriophage-based assay,3D culture and smear were 54.3% & 100%,41.7% & 88.9%,31.7% & 93.5% and 14.6% & 98.9%,respectively.Conclusions TB-RNA is an effective clinical diagnostic method for Mycobacteria tuberculosis.Although the sensitivity of smear is poorer than others,it is a universal testing method in clinical laboratory due to low cost.The positive rate of mycobacteria tuberculosis for 3D culture is lower than that of bacteriophage-based assay and TB-RNA.Although the time to result for 3D culture might last for few weeks,the isolates can be used for drug resistance screening and bacterial identification.

OBJECTIVETo observe the effect of low molecular weight heparin calcium (LMWHC) combined Compound Danshen Injection (DI) on nephrotic syndrome patients with early onset severe preeclampsia.

METHODSTotally 80 nephrotic syndrome patients with early onset severe pre-eclampsia were randomly assigned to four groups voluntarily, i.e., Group A (22 cases, treated by magnesium sulfate), B (19 cases, treated by magnesium sulfate plus LMWHC), C (21 cases, magnesium sulfate plus DI), D (18 cases, magnesium sulfate plus LMWHC and DI). Umbilical arterial S/D ratios, amniotic fluid index (AFI), prolonged gestational age, placenta weight, neonatal weight, and Apgar score were compared among the four groups.

RESULTSCompared with before treatment in the same group, umbilical arterial S/D ratios decreased in the four groups (P <0. 05). AFI decreased in Group A, while it increased in Group B, C, and D (P<0. 05). Compared with Group A at the same time point, umbilical arterial S/D ratios decreased, and AFI increased in Group B, C, and D (P <0. 01 , P <0. 05). Prolonged gestational age and neonatal weight were increased in Group B, C, and D (P <0. 01, P <0. 05). Placenta weight were increased in Group B and D (P <0. 05). Apgar scores at 1 and 5 min were improved in Group D (P <0. 05). Compared with Group B and C at the same time point, umbilical arterial S/D ratios decreased, and AFI increased in Group D (P<0. 05). Compared with Group B, prolonged gestational age and placenta weight were decreased in Group C, but prolonged gestational age and placenta weight were increased in Group D (P <0.05). Compared with Group C, prolonged gestational age, placenta weight, and neonatal weight were increased in Group D (P <0. 05).

CONCLUSIONTreatment of nephrotic syndrome patients with early onset severe pre-eclampsia by LMWHC combined DI could prolong gestational ages, obviously improve prenatal outcomes, with better effect obtained than using any of them alone.

Calcium ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gestational Age ; Heparin, Low-Molecular-Weight ; therapeutic use ; Humans ; Magnesium Sulfate ; Nephrotic Syndrome ; drug therapy ; Phenanthrolines ; Pre-Eclampsia ; drug therapy ; Pregnancy ; Salvia miltiorrhiza

To investigate the effect of Tanreqing injection on immune activity of peripheral blood lymphocytes of patients with lung cancer. The peripheral blood lymphocytes of patients with lung cancer and healthy persons were separated by the density gradient centrifugation method for subsequent experiments, with those from healthy persons as the positive control. The effect of Tanreqing injection on stimulating the proliferation of lymphocytes with phytohemagglutinin (PHA) was determined by MTT method. The effect of Tanreqing injection on the lymphocyte secretions of IFN-γ and TNF-α and the subset ratio of lymphocytes cultured separately or with Tanreqing injection of different concentrations were examined by ELISA and flow cytometry (FCM) respectively. In addition, the LDH release assay was used to detect the cytotoxicity of cytotoxic T cells (CTL) and natural killer cells (NK). According to the findings, all of immunological indexes of lymphocytes from patients with lung cancer were weaker than that of healthy persons, but with the obvious increases in proliferation activity and IFN-γ and TNF-α secretions of lymphocytes co-cultured with Tanreqing Injection (P < 0.05). Among lymphocyte subsets co-cultured with Tanreqing Injection, CD3+, CD3+ CD4+ and CD3- CD16 + 56+ cell ratios notably increased, whereas CD4+ CD25+ Treg cell ratio obviously decreased (P < 0.05). In the meantime, Tanreqing injection can markedly promote the cytotoxicities of CTL and NK (P < 0.05). In conclusion, Tanreqing injection shows a significant effect in promoting the immune activity of lymphocytes from patients with lung cancer and their anti-tumor immunity.
Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interferon-gamma ; genetics ; immunology ; Killer Cells, Natural ; drug effects ; immunology ; Lung Neoplasms ; drug therapy ; genetics ; immunology ; physiopathology ; T-Lymphocytes, Cytotoxic ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; genetics ; immunology

To study the influence of behavioral stress on hippocampal spatial learning and memory, we used the freely moving rats that had undergone chronic implantation of a recording electrode in the hippocampus CA1 region and a bipolar stimulating electrode in the ipsilateral Schaffer collateral-commissural pathway. The field excitatory postsynaptic potentials (fEPSPs) were recorded in the absence of exogenous induction of high-frequency stimulation (HFS) or low-frequency stimulation (LFS) and reflected the effect of stress on the hippocampal spatial learning. And we also investigated the change of hippocampal synaptic plasticity when rats were re-exposed to the same environment at 24 h after novelty acquisition. We found that exploration of a novel environment induced the hippocampal synaptic depression in the rats with stress-adaption, whereas exposure to the novel environment induced the hippocampal synaptic potentiation in the behavioral stress rats. Furthermore, re-exposure to the same environment no longer elicited the hippocampal synaptic potentiation or depression at 24 h after the first novel acquisition in the behavioral stress rats. These results demonstrate that behavioral stress induces the hippocampal synaptic potentiation under novelty acquisition and further damages the hippocampal spatial learning and memory. However, the stress can be adapted by re-exposure to the novelty and thus does not further damage the hippocampal spatial learning and memory.
Animals ; CA1 Region, Hippocampal ; physiology ; Electric Stimulation ; Electrodes, Implanted ; Excitatory Postsynaptic Potentials ; physiology ; Exploratory Behavior ; physiology ; Learning ; physiology ; Male ; Memory ; physiology ; Neuronal Plasticity ; physiology ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; physiology ; Synaptic Potentials ; physiology

OBJECTIVEThis study aimed to investigate the value of child oral health for Chengdu parents, their intentions, and factors influencing their decision to acquire oral insurance coverage for their childrens.

METHODSA total of 562 Chengdu parents were interviewed using questionnaires by convenient sampling, and the results were analyzed using SPSS 20.0.

RESULTSThe age of children (B = -1.741, P = 0.004), age of parents (B = 2.031, P = 0.003), level of oral discomfort (B = 0.569, P = 0.000), incurring/not incurring oral care expenses in the previous year (B = 1.897, P = 0.014), the last time parents' had teeth cleaned (B = 0.777, P = 0.006), and acquiring/not acquiring commercial insurance coverage (B = 1.632, P = 0.031) significantly influenced the intention of acquiring child oral insurance.

CONCLUSIONChild oral health, health and insurance awareness of parents, and other factors influenced the intention of parents to purchase oral insurance coverage for their children, which were significant to establish pediatric dental insurance.

Child ; China ; Dental Care ; Health Expenditures ; Humans ; Insurance, Dental ; economics ; Oral Health ; economics ; Parents ; Surveys and Questionnaires

The effect of magnetic Fe3O4 particles on cellulase in the enzymatic hydrolysis of sunflower seed hull was studied in different adding ways and additive amount. In the process of enzymatic hydrolysis of sunflower seed hull, the variations of cellulase activity, reducing sugar concentration and cellulose conversion were evaluated. After the reaction, the analysis of pH and surface tension of hydrolysate were also used to determine the mechanisms of cellulase by the magnetic effect. The results indicated that after adding magnetic Fe3O4, the cellulase activity, reducing sugar concentration and conversion of cellulose had an increased between the 0.5 g/L and 2.0 g/L cases after 48 h. When the additive amount of magnetic Fe3O4 was 2 g/L, the cellulase activity at 60 h was improved significantly by 25.9%. It was found that the concentration of reducing sugar was increased from 6.950 mg/mL to 8.775 mg/mL with magnetic Fe3O4 1.5 g/L. Simultaneously, compared with the blank, which the conversion of cellulose was 47.932%, the maximum celluloseconversion of samples with adding magnetic Fe3O4 was 60.531%. Besides, the stability of cellulase activity adding in times was better than in one time. After the reaction, the final surface tension of hydrolysate with 1.5 g/L magnetic Fe3O4 was the lowest in comparison with the blank. However, no significant differences were observed in the final pH of the hydrolysate.